Research ArticleEffects of Acupuncture Knife on InflammatoryFactors and Pain in Third Lumbar Vertebrae TransverseProcess Syndrome Model Rats

Jia Ni Yu1 Chang Qing Guo1 Bo Hu2 Nai Gang Liu3 Hong Mei Sun4 Hong Xu4

Hai Xia Wu4 Yan Guo1 Chu Xi Liang1 Zhan Xia Chen1 and Xiao Hong Li1

1 School of Acupuncture and Moxibustion Beijing University of Chinese Medicine No 11 North 3rd Ring East RoadChaoyang District Beijing 100029 China

2 Acupuncture amp Moxibustion and Physiotherapy Department Beijing Shuili Hospital No 19 Yu Yuan Tan South RoadHaidian District Beijing 100036 China

3 Acupuncture and Moxibustion Department China-Japan Friendship Hospital No 2 The Ying Hua Yuan East StreetChaoyang District Beijing 100029 China

4 School of Basic Medical Sciences Beijing University of Chinese Medicine No 11 North 3rd Ring East RoadChaoyang District Beijing 100029 China

Correspondence should be addressed to Chang Qing Guo guochangqing818163com

Received 29 July 2014 Accepted 8 October 2014 Published 2 December 2014

Academic Editor Zhang Weibo

Copyright copy 2014 Jia Ni Yu et alThis is an open access article distributed under the Creative Commons Attribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

The aim of this paper was to explore the long-term effects and pain reliefmechanism of acupuncture knife on third lumbar vertebrae(L3) transverse process syndrome Forty SD rats were randomized into control model electroacupuncture (EA) and acupuncture

knife (AK) group Except control rats other ratswere subjected to an operation to emulate L3transverse process syndrome Fourteen

days after the operation EA and AK rats were given electroacupuncture and acupuncture knife treatments respectively Fifty-sixdays after the operation enzyme-linked immunosorbent assay was used to measure substance P (SP) 5-hydroxytryptamine (5-HT) interleukin-1120573 (IL-1120573) interleukin-10 (IL-10) tumor necrosis factor-120572 (TNF-120572) and transforming growth factor-120573 (TGF-120573)in peripheral blood The tail flick test was used to observe pain threshold We found that rats with the simulation operation hadsignificantly higher levels of SP 5-HT IL-1 IL-10 TNF-120572 and TGF-120573 while the AK rats had lower levels In addition the painthreshold of AK rats was similar to that of control rats AK pretreatment could alleviate pain through modulating inflammatoryresponse

1 Introduction

Low back pain is one of the most commonmedical problemsand is caused by myofascial intervertebral disk (IVD) facetjoint or sacroiliac joint problems The third lumbar vertebra(L3) which adheres to most muscles fascia and other soft

tissues of lumbar is the only vertebra without protection ofthe costal bone or iliac bone However L

3is located in the

most prominent part in the lumbar spine [1] L3transverse

process syndrome is characterized by regional tendernesscaused by overuse and injury of soft tissues adhering to

the transverse process of L3 This syndrome is caused by

peripheral sensitization of local muscle nociceptorsAcupuncture knife invented by ProfessorHanzhang Zhu

is a novel therapy combining traditional Chinese medicinemeridian theory with modern surgical techniques that usesa needle-knife as a therapeutic tool According to previousrandomized controlled multicenter clinical study acupunc-ture knife can restore the dynamic balance and relieve pain byrelaxing and stripping the taut node and cord formed in theL3transverse process tip which is more effective than that of

other medical produces [2 3] Our early stage studies showed

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2014 Article ID 892406 7 pageshttpdxdoiorg1011552014892406

2 Evidence-Based Complementary and Alternative Medicine

that acupuncture knife can adjust analgesic compounds in theshort-term [4 5] However there is no preclinical researchabout the mechanisms of acupuncture knife on long-termanalgesia Substance P (SP) and 5-hydroxytryptamine (5-HT)in the central nervous system and in the periphery have longbeen considered to have an important role in the controlof pain [6 7] In addition animal studies have shown thatinflammatory cytokines contribute to the development ofhyperalgesia [8]

In the present study we aimed to understand themechanism of acupuncture knife on third lumbar vertebraetransverse process syndrome and its effect on substance P(SP) 5-hydroxytryptamine (5-HT) interleukin-1120573 (IL-1120573)interleukin-10 (IL-10) tumor necrosis factor-120572 (TNF-120572) andtransforming growth factor-120573 (TGF-120573) in blood serum

2 Materials and Methods

21 Animals and Surgical Protocol Forty 3-month-old maleSprague-Dawley (SD) rats were used in this study weighing250ndash270 g All rats were purchased from the Victoria-LihuaAnimal Laboratory Center with batch number SCXK (Bei-jing China) 2007-0001

The rats were housed in a controlled environment (12 hlightdark cycle room temperature 23 plusmn 2∘C and 50ndash60relative humidity) with food andwater provided according tothe requirements of the Provision and General Recommen-dations of National Institutes of Health Guide for the Careand Use of Laboratory Animals The Animal Research EthicsBoard of Beijing University of Chinese Medicine approvedthe experiment

Rats were randomly divided into four groups (119899 = 10)a control group in which rats were not given interventiona model group in which rats received surgery to simulateL3transverse process syndrome without treatment an elec-

troacupuncture (EA) group in which rats received surgeryand electroacupuncture treatment 14 days after surgery andan acupuncture knife (AK) group in which rats receivedsurgery and acupuncture knife treatment 14 days aftersurgery The method of establishing L

3transverse process

syndrome model was modified from that of Wang et al [9]Before the surgery the rats were fixed in a prone positionwith10 chloral hydrate (Beijing Factory of Chemical ReagentsBeijing China) abdominal anesthetization (04 gkg) Thehair around the L

3transverse process tip was removed

and a 1 cm vertical incision was made 05ndash08 cm to theleft of the L

3-L4spinal transverse process under aseptic

condition The deep myofascia was separated and the leftlumbar paraspinal muscle was exposed 05ndash08 cm left ofthe central line The paraspinal muscle was separated to theposterior of L

3transverse process and a piece of 05 times 05 cm

absorbent gelatin sponge was implanted (Nanjing JinlingPharmaceutical Co Ltd Nanjing China) After surgerythe lumbar paraspinal muscle of rats was sutured with plaingut suture (3-0) and the cutaneous incision with silk (4-0)The wound was sterilized with gentamicin (2mL 80000UTianjin Pharmaceutical Co Ltd Tianjin China) to avoidinfection

22 Treatment and Techniques For the EA group (119899 = 10)14 days after creating the model intermittent direct EA wasperformed every other day six times in 2 weeks 20mineach time with a frequency of 100Hz dense-disperse waveand 2mA current EA was conducted with the rats lightlyanaesthetized with diethyl ether for about 1min until all fourlimbs were restrained Two points were chosen Yaoyangguan(GV 3) and left Shenshu (BL 23) Disposable stainless sterilehandle needles (Beijing Zhongyan Taihe Medical InstrumentCo Ltd Tianjin China) with a length of 30mm and adiameter of 030mm were used The rats were fixed in theprone position and the acupuncture points were shaved anddisinfected with 75 alcohol Both needles were insertedpercutaneously at the tip of the acupuncture pointsWhen theacupuncturist felt the deqi sensation of the subject the anodelead of the electric stimulator (Hanrsquos 200E Nanjing JishengMedical Co Ltd Jiangsu China) was connected with BL 23and the cathode lead was connected with GV 3

For the AK group (119899 = 10) 14 days after creating themodel acupuncture knife treatment was performed every7 days in 2 weeks two times in total AK was conductedwith the rats lightly anaesthetized with diethyl ether forabout 1min until all four limbs were restrained Pathologicalnodes or cords as acupuncture knife points whichweremuchfirmer than the surrounding muscle fibers were found bygently palpated After acupuncture knife points were shavedand disinfected with 75 alcohol The instrument was usedto make three cuts parallel to the spine and was then rotated90∘ to make another cut The instrument was removed andthe wound was compressed with gauze to avoid excessivebleeding

After treatments were finished all rats were fed foranother 28 days without any treatments or interventions

23 Assessment

231 Pain Threshold At selected time points (all beforenoon) day 0 (the day before model establishment) day 2day 8 day 14 (the day before performing treatment) day 17(1 day after the first acupuncture knife treatment) day 21 day28 (2 days after treatment) day 35 day 42 day 49 and day56 the tail flick test was conducted with a noxious thermalstimulation focus on the site of ventral surface approximately15mm from the tail tip using a radiant heat source providedby a light and thermalgesia algometer (Institute of MaterialMedica Chinese Academy of Medical Sciences BeijingChina) Each rat was held and gently restrained above theapparatus A built-in timer was stopped automatically whenthe tail of the rat withdrew from the beam of light and theresult was displayed for viewing Each rat was tested threetimes with an interval of 15min and the mean of three timeswas the result of pain threshold

232 Transmission Electron Microscopy (TEM) to Observethe Myofibril Ultrastructure When the rats were sacrificeda piece of paraspinal muscle was harvested (1 times 1 times 1mm3)The injured paraspinal muscle tissues were fixed in 5glutaraldehyde for 1ndash3 h washed in buffer and then fixed

Evidence-Based Complementary and Alternative Medicine 3

(a) times12000 (b) times12000

(c) times12000 (d) times12000

Figure 1 Representative ultrastructure changes of skeletal muscles after treatments in the following groups (119899 = 10 per group) (a) control(b) model (c) EA and (d) AK

in 1 osmic acid with pH adjusted to 72ndash74 at 4∘C Thetissues were dehydrated with an ethanol gradient and werethen placed in 100 acetone for 10 minutes Tissue wasthen embedded in a mixture of 100 dehydrating agent andequivalent embedding medium (Epon 812) for 60 minutesbefore they were finally placed in pure embedding mediumovernight at 4∘C The structures of myofibers were analyzedwith an electron microscope (Hitachi Ltd Japan) Tissuesections of 80 nm were prepared and stained with acetic aciduranium saturated aqueous solution followed by lead citrateThe tissues were examined for morphology changes such asarrangement rules of myofibrils position of the sarcomeresarcomere light band (I line) and dark zones (A line) Anyabnormal Z-Lines skeletal muscle cell membranes nucleimitochondria sarcoplasmic reticulum and T-tubules werealso observed

233 Serum Indicators SP 5-HT TGF-120573 IL-1120573 IL-10 andTNF-120572 were tested by ELISA Each blood sample was mixedwith 90 120583L EDTA-Na

2and 120 120583L aprotinin and centrifuged

(4∘C 2000 rpm) for 15min to separate the serum for bio-chemistry and enzyme linked immunosorbent assay (ELISA)

24 Statistical Analysis SPSS 210 statistical analysis soft-ware was used to analyze the data All data are expressedas mean plusmn SD The data of pain threshold was analyzedby nonparametric Kruskal-Wallis test followed by multiplecomparisons between treatment groups by LSD test Afterthe test of normal distribution and homogeneity one-way

ANOVA analysis was used for comparison of serologicalindicators between treatment groups followed by post hocmultiple comparison Statistical significance was set to 119875 lt005 while highly statistical significance was set to 119875 lt 001

3 Results

31 Ultrastructural Changes of Skeletal Muscles after Treat-ments In the control group the structure of sarcomeres waswell aligned with orderly thick and thin filaments A-bankI-bank H-bank M-line and Z-line were clear and regularAround the junction of the A-bank and I-bank there wasa sarcoplasmic reticulum with obvious triads The structureof mitochondria was clear with normal cristae The sar-coplasmic reticulum distribution was regular with integratedmuscle membrane (Figure 1(a)) In the model group therewas excessive empty space between the myofibrilsThe struc-ture of myofibrils and sarcomeres was disordered A-bankI-bank H-bank M-line and Z-line were irregular or notpresent Some thick or thin filaments were fractured Swellingof mitochondria was observed The number of irregularlyarranged collagenous fibers was increased (Figure 1(b)) Inthe EA group the empty space between the myofibrilswas smaller than that in the model group The number offractured filaments was lower than that in the model groupCollagenous fibers were evident (Figure 1(c)) In the AKgroup the empty space between the myofibrils was smallerthan that in the model group A small number of filamentswere fractured The structure of mitochondria was almost

4 Evidence-Based Complementary and Alternative MedicinePa

in th

resh

old

of ta

il fli

ck te

st

Time points (day)

13

12

11

10

9

8

7

6

5

4

0 2 8 14 17 21 28 35 42 49 56

lowast lowastlowast

lowast

lowastlowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

ControlModel

EAAK

Figure 2 Effects of pain threshold on tail flick test in each group(119899 = 10 per group) control model EA and AK Compared withthe control group lowast119875 lt 001 lowastlowast119875 lt 005 Compared with the modelgroup 119875 lt 005

normal Collagenous fibers were observed but arrangementwas orderly (Figure 1(d))

32 Effects of Pain Threshold on Tail Flick Test The results ofthe pain threshold of tail flick test are shown in Figure 2 Thepain threshold of the control group at different time pointsdid not showobvious changesTheother three groups showedlower pain threshold from the second day after the operationAt 14 days the day before performing treatment comparedwith the control group the model EA and AK group had alower pain threshold (119875 lt 001 or119875 lt 005) and therewere nostatistical differences between themodel EA andAK groupsAfter treatments the pain threshold of the EA and AK grouprose Fromdays 17 to 28 comparedwith the control group thepain thresholds of the EA andAKgroupswere not statisticallydifferentHowever the pain threshold of themodel groupwassignificantly lower than the control group On days 21 and 28compared with the model group the pain threshold of theAK group rose significantly (119875 lt 005) From days 35 to 56there was no significant difference between all four groupsHowever Figure 1 shows a similar pain threshold between thecontrol EA and AK group which is higher than that of themodel group

33 Changes in IL-1120573 IL-10 TNF-120572 and TGF-120573 in thePeripheral Blood Fifty-six days after operation IL-1120573 andTNF-120572 levels in the model group were significantly higherthan that in the other three groups (119875 lt 005 or 119875 lt 001)Compared with the control group there was no significantdifference in IL-1120573 and TNF-120572 in the EA and AK groups(119875 gt 005) There was no significant difference in IL-1120573 andTNF-120572 between the EA and AK groups (119875 gt 005) (Figure 3)The IL-10 level in the model group was significantly higher

lowast

lowast

ControlModel

EAAK

8

6

4

2

0

Seru

m v

alue

of I

L-1120573

and

TNF-120572

(ng

mL)

IL-1120573 TNF-120572

Figure 3 Effects of AK on IL-1120573 and TNF-120572 in peripheral blood inthe following groups (119899 = 10 per group) control model EA andAK Compared with the control group lowast119875 lt 001 compared withthe model group 119875 lt 001 119875 lt 005

lowast

lowast

lowastlowastlowast

Seru

m v

alue

of I

L-10

and

TGF-120573

IL-10 (ngmL) TGF-120573 (pgmL)

350

300

250

200

150

100

50

0

ControlModel

EAAK

Figure 4 Effects of AK on IL-10 and TGF-120573 in peripheral bloodin the following groups (119899 = 10 per group) control modelEA and AK Compared with control group lowast119875 lt 001 lowastlowast119875 lt 005compared with model group 119875 lt 001

than that in the other three groups (119875 lt 001) Comparedwith the control group there was no significant differencein IL-10 in the EA and AK groups (119875 gt 005) There wereno significant differences in IL-10 among the EA and AKgroups (119875 gt 005) Compared with the control group TGF-120573 in the model EA and AK groups were significantly higher(119875 lt 001 or 119875 lt 005) Compared with the model groupTGF-120573 in the EA and AK groups were significantly lower(119875 lt 001) There was no difference between the EA and AKgroups (119875 gt 005 Figure 4)

Evidence-Based Complementary and Alternative Medicine 5

ControlModel

EAAK

lowast

lowast

lowastlowast

lowast300

270

240

210

180

150

120

90

60

30

0

Seru

m v

alue

of S

P an

d5

-HT

SP (pgmL) 5-HT (ngmL)

lowast

lowast

Figure 5 Effects of AK on SP and 5-HT in peripheral blood in thefollowing groups (119899 = 10 per group) control model EA and AKCompared with the control group lowast119875 lt 001 lowastlowast119875 lt 005 comparedwith themodel group 119875 lt 001 comparedwith the EA group 998771119875 lt005

34 Changes in SP and 5-Hydroxytryptamine in the PeripheralBlood Fifty-six days after operation compared with the con-trol group the model EA and AK groups had significantlyhigher SP levels in the peripheral blood (119875 lt 005 or 119875 lt001) Compared with the model group the EA and AKgroups had significantly lower SP levels in the peripheralblood (119875 lt 001) There was no significant difference inSP among the EA and AK groups (119875 gt 005) Comparedwith the control group the model EA and AK groups hadsignificantly higher 5-HT levels in the peripheral blood (119875 lt001) Compared with the model group the EA and AKgroups had significantly lower 5-HT levels in the peripheralblood (119875 lt 001) The 5-HT of the AK group was higher thanthat of the EA group (119875 lt 005 Figure 5)

4 Discussion

The term ldquotendon meridianrdquo in traditional Chinese medicineis part of the meridian system with functions of connectinglimbs and regulating the movement of joints Qi and bloodin meridians nourish some tissues involved in movementsuch as tendons ligaments fascia skeletal muscles andjoint capsules Some typical symptoms of tendon meridiandisease are pain numbness hypertonicity or stiffness ofmuscle or fascia even motor dysfunction which are possiblyequivalent to chronic injury of the motor system Thirdlumbar vertebrae transverse process syndrome is part of lowback pain according to TCM theory which is commonlybecause of qi and blood stagnation in the waist Disorderedblood circulation in the waist is caused by acute trauma achronic strain or invasion of cold pathogens The syndromeincludes spontaneous unilateral regional pain near the thirdlumbar vertebrae transverse process or referred pain tothe homolateral leg that is aggravated by bending down

with restricted range of waist motion and the presence ofhypersensitive nodules and local tenderness near the thirdlumbar vertebrae transverse process

After acute or chronic injury soft tissue adheres tothe tip of the third lumbar vertebrae transverse processwhich causes aseptic inflammation Inflammation causespathological changes such as edema exudation adhesion oreven continuous contracture in lesion tissues Nociceptiveterminals in muscle have a multitude of different receptors intheir membranes including matched receptors for moleculesthat are released from damaged tissue such as bradykininserotonin and prostaglandins [10] The continuous presenceof these inflammatorymediators and other chemicals may benecessary for persistent pain conditions [10]

Cytokine TNF-120572 is a mediator of chronic pain producedby the onset of inflammation [11] TNF-120572 can lead to NF-120581B-mediated transcription of IL-1 whichmediates inflammatoryhyperalgesia [12] IL-1120573 is one of the most important mem-bers of the interleukin-1 family When IL-1120573 binds to the IL-1 receptor the activation of IL-1 receptors induces changesin synaptic strength and results in behavioral hyperalgesia[13] Our results showed that serum TNF-120572 and IL-1120573 in themodel rats remained at a higher level than that in the normalrats This implies that the operation caused chronic inflam-mation The increased cytokines confirmed the activation ofimmune cells and the production of sterile inflammationThe rat model of embedding absorbent gelatin is suitablefor the observation of chronic inflammation and pain Afterelectroacupuncture and acupuncture knife treatments thelevels of serumTNF-120572 and IL-1120573were significantly lower thanthose in model rats Therefore both the electroacupunctureand acupuncture knife treatments can reduce the releaseof cytokines In addition the effect of anti-inflammation islong term Compared with EA rats there were no significantdifferences in AK rats in both serum levels of TNF-120572 andIL-1120573 However the contents of TNF-120572 and IL-1120573 in AKrats were closer to those in normal rats which suggeststhat acupuncture knife treatment is a more effective anti-inflammatory than electroacupuncture

IL-10 is a powerful anti-inflammatory cytokine that canreduce the production of proinflammatory cytokines suchas IL-1120573 IL-6 TNF-120572 and nerve growth factor IL-10-producing monocytes and macrophages promote the reso-lution of transient inflammatory hyperalgesia Impaired IL-10 production may represent a risk for developing chronicpain after inflammation [14] Moreover IL-10 gene therapyhas been proven useful in neuropathic pain rats [15] Ourresults showed that the serum IL-10 levels in model EA andAK rats were significantly higher than those in normal ratsHowever compared with model rats the serum levels in EArats and AK rats were significantly lower This result mayprove the therapeutic effect of downregulation of proinflam-matory cytokines by electroacupuncture and acupunctureknife treatments

TGF-120573 has been recognized as a growth factor withmultiple biological functions TGF-120573 plays an important rolein the regulation of immune cell function by limiting the pro-duction of inflammatory mediators including interleukin-2 and tumor necrosis factor [16] In chronic inflammation

6 Evidence-Based Complementary and Alternative Medicine

TGF-120573 is expressed and promotes wound healing and fibrosis[17] In addition TGF-120573 promotes the expression of endoge-nous opioids inhibiting the neuroimmune responses of glialcells and neurons in the spinal cord following peripheralinjuries Other undetermined mechanisms also contribute toits antiallodynic effect [18] Our results showed that the serumlevels of TGF-120573 in model EA and AK rats were significantlyhigher than those in normal rats which implies that theperipheral injury tissue is under the fibroplasia stage Thiswas also shown by pathological observations In addition thecontent of TGF-120573 in EA and AK rats is significantly lowerthan that in model rats This demonstrates that the level ofTGF-120573 declined because of the lighter inflammatory responsebrought by EA and AK treatments

Substance P (SP) is a neuropeptide released from nerveendings in many tissues which is directly related with painand plays an important role in inflammation [19] Peripheralinflammation and noxious stimuli may trigger the releaseof SP [20] thus lowering pain thresholds and resultingin allodynia [21] In the periphery administration of 5-HT can increase the excitability of myelinated (A120575 fibers)and unmyelinated afferents (C fibers) [22] Previous studyconfirmed that the inhibition of 5-HT secretion from mastcells can diminish the sensitization of sensory nerve terminals[23] Our results showed that 56 days after operation theconcentration of SP and 5-HT in the blood of EA and AKrats was lower than that in model rats and higher than thatin normal rats Therefore both EA and AK treatments con-tribute to relieving painThis may be because of alleviation oflocal inflammatory responses However compared with AKrats serum 5-HT of EA rats is significantly lowerThis impliesthat EA treatment is more effective for alleviating pain

5 Conclusions

In summary we demonstrated that acupuncture knife ther-apy is able to mitigate pain by controlling the inflammatoryresponse and is capable of restoring injury to soft tissueHowever the analgesic effect of electroacupuncture therapyis better than that of acupuncture knife therapy This maybe because of fewer treatment sessions for acupuncture knifetherapy (two times) than electroacupuncture therapy (sixtimes) accompanied by retaining the needle for 20minHowever acupuncture knife therapy has a better trendfor reducing inflammation In addition in spite of widercutting edge of acupuncture knife than acupuncture needlethe puncture depth of acupuncture knife is shallower thanacupuncture needle that is cutting in taut nodes and cordsof muscle or fascia located under skin without arteries veinsor nerves which can lead to irreversible damages The safetyof acupuncture knife is credible

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors show their deepest gratitude to ProfessorHanzhang Zhu for inventing the acupuncture knife treat-ment They thank him for his contribution to Chinesemedicine

References

[1] C M Ni A Zhao and J J He ldquoThe relationship betweenthe third lumbar transverse process syndrome and anatomyrdquoJournal of AnhuiMedical University vol 27 no 1 pp 11ndash13 1992

[2] C Q Guo S L Li J L Qiao et al ldquoMulticenter and randomizedcontrolled clinical trial of acupuncture knife therapy in treat-ment of third lumbar transverse process syndromerdquo Journal ofChengduUniversity of ChineseMedicine vol 35 no 1 pp 20ndash232012

[3] C Q Guo F H Dong S L Li et al ldquoEffects of acupotomy lysison local soft tissue tension in patients with the third lumbarvertebrae transverse process syndromerdquoChinese acupuncture ampMoxibustion vol 32 no 7 pp 617ndash620 2012

[4] B Hu L Liu C-Q Guo et al ldquoEffect of acupotomy lysis on SPand CCK-8 contents in hypothalamus and spinal cord in ratswith transverse process syndrome of the third lumbar vertebrardquoAcupuncture Research vol 33 no 1 pp 22ndash25 2008

[5] H M Sun B Hu X H Li et al ldquoEffect of acupuncture knifedissolution on pain threshold and ENK and 120573-EP in spinalcord and hypothalamus in rats with transverse syndrome of thethird lumbar vertebraerdquo Journal of Beijing University of ChineseMedicine vol 31 no 3 pp 176ndash179 2008

[6] E D de Avila R S de Molon D A de Godoi Goncalves andC M Camparis ldquoRelationship between levels of neuropeptideSubstance P in periodontal disease and chronic pain a literaturereviewrdquo Journal of Investigative and Clinical Dentistry vol 5 no2 pp 91ndash97 2014

[7] L Bardin ldquoThe complex role of serotonin and 5-HT receptorsin chronic painrdquo Behavioural Pharmacology vol 22 no 5-6 pp390ndash404 2011

[8] M de Goeij L T Van Eijk P Vanelderen et al ldquoSystemicinflammation decreases pain threshold in humans in vivordquoPLoS ONE vol 8 no 12 Article ID e84159 2013

[9] J RWang J L Qiao P Lu et al ldquoConstruct and research of ani-mal model for the third lumbar transverse process syndromerdquoChinese Journal of Physical Medicine and Rehabilitation vol 25no 7 pp 394ndash398 2003

[10] J P Shah J V Danoff M J Desai et al ldquoMochemicalsassociated with pain and inflammation are elevated in sites nearto and remote from active myofascial trigger pointsrdquo Archivesof Physical Medicine and Rehabilitation vol 89 no 1 pp 16ndash232008

[11] Q Liu L Jin B H Mahon M D Chordia F H Shen and XLi ldquoA novel treatment of neuroinflammation against low backpain by soluble fullerol nanoparticlesrdquo Spine vol 38 no 17 pp1443ndash1451 2013

[12] C A Dinarello A Simon and J W M van der Meer ldquoTreatinginflammation by blocking interleukin-1 in a broad spectrum ofdiseasesrdquoNature Reviews Drug Discovery vol 11 no 8 pp 633ndash652 2012

[13] K Ren and R Dubner ldquoInteractions between the immune andnervous systems in painrdquo Nature Medicine vol 16 no 11 pp1267ndash1276 2010

Evidence-Based Complementary and Alternative Medicine 7

[14] H L D M Willemen N Eijkelkamp A Garza Carbajal etal ldquoMonocytesmacrophages control resolution of transientinflammatory painrdquo Journal of Pain vol 15 no 5 pp 496ndash5062014

[15] E D Milligan E M Sloane S J Langer et al ldquoRepeatedintrathecal injections of plasmid DNA encoding interleukin-10produce prolonged reversal of neuropathic painrdquo Pain vol 126no 1ndash3 pp 294ndash308 2006

[16] N F Chen S Y Huang W F Chen et al ldquoTGF-1205731 attenuatesspinal neuroinflammation and the excitatory amino acid systemin rats with neuropathic painrdquo The Journal of Pain vol 14 no12 pp 1671ndash1685 2013

[17] Y Zhu T Colak M Shenoy et al ldquoTransforming growthfactor beta induces sensory neuronal hyperexcitability andcontributes to pancreatic pain and hyperalgesia in rats withchronic pancreatitisrdquoMolecular Pain vol 8 article 65 2012

[18] A Lantero M Tramullas A Diaz and M A Hurle ldquoTrans-forming growth factor-120573 in normal nociceptive processing andpathological pain modelsrdquoMolecular Neurobiology vol 45 no1 pp 76ndash86 2012

[19] M Munoz and R Covenas ldquoInvolvement of substance P andthe NK-1 receptor in human pathologyrdquo Amino Acids vol 46no 7 pp 1725ndash1750 2014

[20] B Bie and Z-Q Zhao ldquoPeripheral inflammation alters desen-sitization of substance P-evoked current in rat dorsal rootganglion neuronsrdquo European Journal of Pharmacology vol 670no 2-3 pp 495ndash499 2011

[21] K L Witt and J A Vilensky ldquoThe anatomy of osteoarthriticjoint painrdquo Clinical Anatomy vol 27 no 3 pp 451ndash454 2014

[22] L Bardin ldquoThe complex role of serotonin and 5-HT receptors inchronic painrdquo Behavior Pharmacollgy vol 22 no 5-6 pp 390ndash404 2011

[23] I Ferjan and M Lipnik-Stangelj ldquoChronic pain treatment Theinfluence of tricyclic antidepressants on serotonin release anduptake in mast cellsrdquo Mediators of Inflammation vol 2013Article ID 340473 7 pages 2013

Evidence-Based Complementary and Alternative Medicine 3

(a) times12000 (b) times12000

(c) times12000 (d) times12000

Figure 1 Representative ultrastructure changes of skeletal muscles after treatments in the following groups (119899 = 10 per group) (a) control(b) model (c) EA and (d) AK

in 1 osmic acid with pH adjusted to 72ndash74 at 4∘C Thetissues were dehydrated with an ethanol gradient and werethen placed in 100 acetone for 10 minutes Tissue wasthen embedded in a mixture of 100 dehydrating agent andequivalent embedding medium (Epon 812) for 60 minutesbefore they were finally placed in pure embedding mediumovernight at 4∘C The structures of myofibers were analyzedwith an electron microscope (Hitachi Ltd Japan) Tissuesections of 80 nm were prepared and stained with acetic aciduranium saturated aqueous solution followed by lead citrateThe tissues were examined for morphology changes such asarrangement rules of myofibrils position of the sarcomeresarcomere light band (I line) and dark zones (A line) Anyabnormal Z-Lines skeletal muscle cell membranes nucleimitochondria sarcoplasmic reticulum and T-tubules werealso observed

233 Serum Indicators SP 5-HT TGF-120573 IL-1120573 IL-10 andTNF-120572 were tested by ELISA Each blood sample was mixedwith 90 120583L EDTA-Na

2and 120 120583L aprotinin and centrifuged

(4∘C 2000 rpm) for 15min to separate the serum for bio-chemistry and enzyme linked immunosorbent assay (ELISA)

24 Statistical Analysis SPSS 210 statistical analysis soft-ware was used to analyze the data All data are expressedas mean plusmn SD The data of pain threshold was analyzedby nonparametric Kruskal-Wallis test followed by multiplecomparisons between treatment groups by LSD test Afterthe test of normal distribution and homogeneity one-way

ANOVA analysis was used for comparison of serologicalindicators between treatment groups followed by post hocmultiple comparison Statistical significance was set to 119875 lt005 while highly statistical significance was set to 119875 lt 001

3 Results

31 Ultrastructural Changes of Skeletal Muscles after Treat-ments In the control group the structure of sarcomeres waswell aligned with orderly thick and thin filaments A-bankI-bank H-bank M-line and Z-line were clear and regularAround the junction of the A-bank and I-bank there wasa sarcoplasmic reticulum with obvious triads The structureof mitochondria was clear with normal cristae The sar-coplasmic reticulum distribution was regular with integratedmuscle membrane (Figure 1(a)) In the model group therewas excessive empty space between the myofibrilsThe struc-ture of myofibrils and sarcomeres was disordered A-bankI-bank H-bank M-line and Z-line were irregular or notpresent Some thick or thin filaments were fractured Swellingof mitochondria was observed The number of irregularlyarranged collagenous fibers was increased (Figure 1(b)) Inthe EA group the empty space between the myofibrilswas smaller than that in the model group The number offractured filaments was lower than that in the model groupCollagenous fibers were evident (Figure 1(c)) In the AKgroup the empty space between the myofibrils was smallerthan that in the model group A small number of filamentswere fractured The structure of mitochondria was almost

4 Evidence-Based Complementary and Alternative MedicinePa

in th

resh

old

of ta

il fli

ck te

st

Time points (day)

13

12

11

10

9

8

7

6

5

4

0 2 8 14 17 21 28 35 42 49 56

lowast lowastlowast

lowast

lowastlowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

ControlModel

EAAK

Figure 2 Effects of pain threshold on tail flick test in each group(119899 = 10 per group) control model EA and AK Compared withthe control group lowast119875 lt 001 lowastlowast119875 lt 005 Compared with the modelgroup 119875 lt 005

normal Collagenous fibers were observed but arrangementwas orderly (Figure 1(d))

32 Effects of Pain Threshold on Tail Flick Test The results ofthe pain threshold of tail flick test are shown in Figure 2 Thepain threshold of the control group at different time pointsdid not showobvious changesTheother three groups showedlower pain threshold from the second day after the operationAt 14 days the day before performing treatment comparedwith the control group the model EA and AK group had alower pain threshold (119875 lt 001 or119875 lt 005) and therewere nostatistical differences between themodel EA andAK groupsAfter treatments the pain threshold of the EA and AK grouprose Fromdays 17 to 28 comparedwith the control group thepain thresholds of the EA andAKgroupswere not statisticallydifferentHowever the pain threshold of themodel groupwassignificantly lower than the control group On days 21 and 28compared with the model group the pain threshold of theAK group rose significantly (119875 lt 005) From days 35 to 56there was no significant difference between all four groupsHowever Figure 1 shows a similar pain threshold between thecontrol EA and AK group which is higher than that of themodel group

33 Changes in IL-1120573 IL-10 TNF-120572 and TGF-120573 in thePeripheral Blood Fifty-six days after operation IL-1120573 andTNF-120572 levels in the model group were significantly higherthan that in the other three groups (119875 lt 005 or 119875 lt 001)Compared with the control group there was no significantdifference in IL-1120573 and TNF-120572 in the EA and AK groups(119875 gt 005) There was no significant difference in IL-1120573 andTNF-120572 between the EA and AK groups (119875 gt 005) (Figure 3)The IL-10 level in the model group was significantly higher

lowast

lowast

ControlModel

EAAK

8

6

4

2

0

Seru

m v

alue

of I

L-1120573

and

TNF-120572

(ng

mL)

IL-1120573 TNF-120572

Figure 3 Effects of AK on IL-1120573 and TNF-120572 in peripheral blood inthe following groups (119899 = 10 per group) control model EA andAK Compared with the control group lowast119875 lt 001 compared withthe model group 119875 lt 001 119875 lt 005

lowast

lowast

lowastlowastlowast

Seru

m v

alue

of I

L-10

and

TGF-120573

IL-10 (ngmL) TGF-120573 (pgmL)

350

300

250

200

150

100

50

0

ControlModel

EAAK

Figure 4 Effects of AK on IL-10 and TGF-120573 in peripheral bloodin the following groups (119899 = 10 per group) control modelEA and AK Compared with control group lowast119875 lt 001 lowastlowast119875 lt 005compared with model group 119875 lt 001

than that in the other three groups (119875 lt 001) Comparedwith the control group there was no significant differencein IL-10 in the EA and AK groups (119875 gt 005) There wereno significant differences in IL-10 among the EA and AKgroups (119875 gt 005) Compared with the control group TGF-120573 in the model EA and AK groups were significantly higher(119875 lt 001 or 119875 lt 005) Compared with the model groupTGF-120573 in the EA and AK groups were significantly lower(119875 lt 001) There was no difference between the EA and AKgroups (119875 gt 005 Figure 4)

Evidence-Based Complementary and Alternative Medicine 5

ControlModel

EAAK

lowast

lowast

lowastlowast

lowast300

270

240

210

180

150

120

90

60

30

0

Seru

m v

alue

of S

P an

d5

-HT

SP (pgmL) 5-HT (ngmL)

lowast

lowast

Figure 5 Effects of AK on SP and 5-HT in peripheral blood in thefollowing groups (119899 = 10 per group) control model EA and AKCompared with the control group lowast119875 lt 001 lowastlowast119875 lt 005 comparedwith themodel group 119875 lt 001 comparedwith the EA group 998771119875 lt005

34 Changes in SP and 5-Hydroxytryptamine in the PeripheralBlood Fifty-six days after operation compared with the con-trol group the model EA and AK groups had significantlyhigher SP levels in the peripheral blood (119875 lt 005 or 119875 lt001) Compared with the model group the EA and AKgroups had significantly lower SP levels in the peripheralblood (119875 lt 001) There was no significant difference inSP among the EA and AK groups (119875 gt 005) Comparedwith the control group the model EA and AK groups hadsignificantly higher 5-HT levels in the peripheral blood (119875 lt001) Compared with the model group the EA and AKgroups had significantly lower 5-HT levels in the peripheralblood (119875 lt 001) The 5-HT of the AK group was higher thanthat of the EA group (119875 lt 005 Figure 5)

4 Discussion

The term ldquotendon meridianrdquo in traditional Chinese medicineis part of the meridian system with functions of connectinglimbs and regulating the movement of joints Qi and bloodin meridians nourish some tissues involved in movementsuch as tendons ligaments fascia skeletal muscles andjoint capsules Some typical symptoms of tendon meridiandisease are pain numbness hypertonicity or stiffness ofmuscle or fascia even motor dysfunction which are possiblyequivalent to chronic injury of the motor system Thirdlumbar vertebrae transverse process syndrome is part of lowback pain according to TCM theory which is commonlybecause of qi and blood stagnation in the waist Disorderedblood circulation in the waist is caused by acute trauma achronic strain or invasion of cold pathogens The syndromeincludes spontaneous unilateral regional pain near the thirdlumbar vertebrae transverse process or referred pain tothe homolateral leg that is aggravated by bending down

with restricted range of waist motion and the presence ofhypersensitive nodules and local tenderness near the thirdlumbar vertebrae transverse process

After acute or chronic injury soft tissue adheres tothe tip of the third lumbar vertebrae transverse processwhich causes aseptic inflammation Inflammation causespathological changes such as edema exudation adhesion oreven continuous contracture in lesion tissues Nociceptiveterminals in muscle have a multitude of different receptors intheir membranes including matched receptors for moleculesthat are released from damaged tissue such as bradykininserotonin and prostaglandins [10] The continuous presenceof these inflammatorymediators and other chemicals may benecessary for persistent pain conditions [10]

Cytokine TNF-120572 is a mediator of chronic pain producedby the onset of inflammation [11] TNF-120572 can lead to NF-120581B-mediated transcription of IL-1 whichmediates inflammatoryhyperalgesia [12] IL-1120573 is one of the most important mem-bers of the interleukin-1 family When IL-1120573 binds to the IL-1 receptor the activation of IL-1 receptors induces changesin synaptic strength and results in behavioral hyperalgesia[13] Our results showed that serum TNF-120572 and IL-1120573 in themodel rats remained at a higher level than that in the normalrats This implies that the operation caused chronic inflam-mation The increased cytokines confirmed the activation ofimmune cells and the production of sterile inflammationThe rat model of embedding absorbent gelatin is suitablefor the observation of chronic inflammation and pain Afterelectroacupuncture and acupuncture knife treatments thelevels of serumTNF-120572 and IL-1120573were significantly lower thanthose in model rats Therefore both the electroacupunctureand acupuncture knife treatments can reduce the releaseof cytokines In addition the effect of anti-inflammation islong term Compared with EA rats there were no significantdifferences in AK rats in both serum levels of TNF-120572 andIL-1120573 However the contents of TNF-120572 and IL-1120573 in AKrats were closer to those in normal rats which suggeststhat acupuncture knife treatment is a more effective anti-inflammatory than electroacupuncture

IL-10 is a powerful anti-inflammatory cytokine that canreduce the production of proinflammatory cytokines suchas IL-1120573 IL-6 TNF-120572 and nerve growth factor IL-10-producing monocytes and macrophages promote the reso-lution of transient inflammatory hyperalgesia Impaired IL-10 production may represent a risk for developing chronicpain after inflammation [14] Moreover IL-10 gene therapyhas been proven useful in neuropathic pain rats [15] Ourresults showed that the serum IL-10 levels in model EA andAK rats were significantly higher than those in normal ratsHowever compared with model rats the serum levels in EArats and AK rats were significantly lower This result mayprove the therapeutic effect of downregulation of proinflam-matory cytokines by electroacupuncture and acupunctureknife treatments

TGF-120573 has been recognized as a growth factor withmultiple biological functions TGF-120573 plays an important rolein the regulation of immune cell function by limiting the pro-duction of inflammatory mediators including interleukin-2 and tumor necrosis factor [16] In chronic inflammation

6 Evidence-Based Complementary and Alternative Medicine

TGF-120573 is expressed and promotes wound healing and fibrosis[17] In addition TGF-120573 promotes the expression of endoge-nous opioids inhibiting the neuroimmune responses of glialcells and neurons in the spinal cord following peripheralinjuries Other undetermined mechanisms also contribute toits antiallodynic effect [18] Our results showed that the serumlevels of TGF-120573 in model EA and AK rats were significantlyhigher than those in normal rats which implies that theperipheral injury tissue is under the fibroplasia stage Thiswas also shown by pathological observations In addition thecontent of TGF-120573 in EA and AK rats is significantly lowerthan that in model rats This demonstrates that the level ofTGF-120573 declined because of the lighter inflammatory responsebrought by EA and AK treatments

Substance P (SP) is a neuropeptide released from nerveendings in many tissues which is directly related with painand plays an important role in inflammation [19] Peripheralinflammation and noxious stimuli may trigger the releaseof SP [20] thus lowering pain thresholds and resultingin allodynia [21] In the periphery administration of 5-HT can increase the excitability of myelinated (A120575 fibers)and unmyelinated afferents (C fibers) [22] Previous studyconfirmed that the inhibition of 5-HT secretion from mastcells can diminish the sensitization of sensory nerve terminals[23] Our results showed that 56 days after operation theconcentration of SP and 5-HT in the blood of EA and AKrats was lower than that in model rats and higher than thatin normal rats Therefore both EA and AK treatments con-tribute to relieving painThis may be because of alleviation oflocal inflammatory responses However compared with AKrats serum 5-HT of EA rats is significantly lowerThis impliesthat EA treatment is more effective for alleviating pain

5 Conclusions

In summary we demonstrated that acupuncture knife ther-apy is able to mitigate pain by controlling the inflammatoryresponse and is capable of restoring injury to soft tissueHowever the analgesic effect of electroacupuncture therapyis better than that of acupuncture knife therapy This maybe because of fewer treatment sessions for acupuncture knifetherapy (two times) than electroacupuncture therapy (sixtimes) accompanied by retaining the needle for 20minHowever acupuncture knife therapy has a better trendfor reducing inflammation In addition in spite of widercutting edge of acupuncture knife than acupuncture needlethe puncture depth of acupuncture knife is shallower thanacupuncture needle that is cutting in taut nodes and cordsof muscle or fascia located under skin without arteries veinsor nerves which can lead to irreversible damages The safetyof acupuncture knife is credible

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors show their deepest gratitude to ProfessorHanzhang Zhu for inventing the acupuncture knife treat-ment They thank him for his contribution to Chinesemedicine

References

[1] C M Ni A Zhao and J J He ldquoThe relationship betweenthe third lumbar transverse process syndrome and anatomyrdquoJournal of AnhuiMedical University vol 27 no 1 pp 11ndash13 1992

[2] C Q Guo S L Li J L Qiao et al ldquoMulticenter and randomizedcontrolled clinical trial of acupuncture knife therapy in treat-ment of third lumbar transverse process syndromerdquo Journal ofChengduUniversity of ChineseMedicine vol 35 no 1 pp 20ndash232012

[3] C Q Guo F H Dong S L Li et al ldquoEffects of acupotomy lysison local soft tissue tension in patients with the third lumbarvertebrae transverse process syndromerdquoChinese acupuncture ampMoxibustion vol 32 no 7 pp 617ndash620 2012

[4] B Hu L Liu C-Q Guo et al ldquoEffect of acupotomy lysis on SPand CCK-8 contents in hypothalamus and spinal cord in ratswith transverse process syndrome of the third lumbar vertebrardquoAcupuncture Research vol 33 no 1 pp 22ndash25 2008

[5] H M Sun B Hu X H Li et al ldquoEffect of acupuncture knifedissolution on pain threshold and ENK and 120573-EP in spinalcord and hypothalamus in rats with transverse syndrome of thethird lumbar vertebraerdquo Journal of Beijing University of ChineseMedicine vol 31 no 3 pp 176ndash179 2008

[6] E D de Avila R S de Molon D A de Godoi Goncalves andC M Camparis ldquoRelationship between levels of neuropeptideSubstance P in periodontal disease and chronic pain a literaturereviewrdquo Journal of Investigative and Clinical Dentistry vol 5 no2 pp 91ndash97 2014

[7] L Bardin ldquoThe complex role of serotonin and 5-HT receptorsin chronic painrdquo Behavioural Pharmacology vol 22 no 5-6 pp390ndash404 2011

[8] M de Goeij L T Van Eijk P Vanelderen et al ldquoSystemicinflammation decreases pain threshold in humans in vivordquoPLoS ONE vol 8 no 12 Article ID e84159 2013

[9] J RWang J L Qiao P Lu et al ldquoConstruct and research of ani-mal model for the third lumbar transverse process syndromerdquoChinese Journal of Physical Medicine and Rehabilitation vol 25no 7 pp 394ndash398 2003

[10] J P Shah J V Danoff M J Desai et al ldquoMochemicalsassociated with pain and inflammation are elevated in sites nearto and remote from active myofascial trigger pointsrdquo Archivesof Physical Medicine and Rehabilitation vol 89 no 1 pp 16ndash232008

[11] Q Liu L Jin B H Mahon M D Chordia F H Shen and XLi ldquoA novel treatment of neuroinflammation against low backpain by soluble fullerol nanoparticlesrdquo Spine vol 38 no 17 pp1443ndash1451 2013

[12] C A Dinarello A Simon and J W M van der Meer ldquoTreatinginflammation by blocking interleukin-1 in a broad spectrum ofdiseasesrdquoNature Reviews Drug Discovery vol 11 no 8 pp 633ndash652 2012

[13] K Ren and R Dubner ldquoInteractions between the immune andnervous systems in painrdquo Nature Medicine vol 16 no 11 pp1267ndash1276 2010

Evidence-Based Complementary and Alternative Medicine 7

[14] H L D M Willemen N Eijkelkamp A Garza Carbajal etal ldquoMonocytesmacrophages control resolution of transientinflammatory painrdquo Journal of Pain vol 15 no 5 pp 496ndash5062014

[15] E D Milligan E M Sloane S J Langer et al ldquoRepeatedintrathecal injections of plasmid DNA encoding interleukin-10produce prolonged reversal of neuropathic painrdquo Pain vol 126no 1ndash3 pp 294ndash308 2006

[16] N F Chen S Y Huang W F Chen et al ldquoTGF-1205731 attenuatesspinal neuroinflammation and the excitatory amino acid systemin rats with neuropathic painrdquo The Journal of Pain vol 14 no12 pp 1671ndash1685 2013

[17] Y Zhu T Colak M Shenoy et al ldquoTransforming growthfactor beta induces sensory neuronal hyperexcitability andcontributes to pancreatic pain and hyperalgesia in rats withchronic pancreatitisrdquoMolecular Pain vol 8 article 65 2012

[18] A Lantero M Tramullas A Diaz and M A Hurle ldquoTrans-forming growth factor-120573 in normal nociceptive processing andpathological pain modelsrdquoMolecular Neurobiology vol 45 no1 pp 76ndash86 2012

[19] M Munoz and R Covenas ldquoInvolvement of substance P andthe NK-1 receptor in human pathologyrdquo Amino Acids vol 46no 7 pp 1725ndash1750 2014

[20] B Bie and Z-Q Zhao ldquoPeripheral inflammation alters desen-sitization of substance P-evoked current in rat dorsal rootganglion neuronsrdquo European Journal of Pharmacology vol 670no 2-3 pp 495ndash499 2011

[21] K L Witt and J A Vilensky ldquoThe anatomy of osteoarthriticjoint painrdquo Clinical Anatomy vol 27 no 3 pp 451ndash454 2014

[22] L Bardin ldquoThe complex role of serotonin and 5-HT receptors inchronic painrdquo Behavior Pharmacollgy vol 22 no 5-6 pp 390ndash404 2011

[23] I Ferjan and M Lipnik-Stangelj ldquoChronic pain treatment Theinfluence of tricyclic antidepressants on serotonin release anduptake in mast cellsrdquo Mediators of Inflammation vol 2013Article ID 340473 7 pages 2013

4 Evidence-Based Complementary and Alternative MedicinePa

in th

resh

old

of ta

il fli

ck te

st

Time points (day)

13

12

11

10

9

8

7

6

5

4

0 2 8 14 17 21 28 35 42 49 56

lowast lowastlowast

lowast

lowastlowastlowastlowastlowast

lowastlowastlowastlowast

lowastlowast

lowastlowast

ControlModel

EAAK

Figure 2 Effects of pain threshold on tail flick test in each group(119899 = 10 per group) control model EA and AK Compared withthe control group lowast119875 lt 001 lowastlowast119875 lt 005 Compared with the modelgroup 119875 lt 005

normal Collagenous fibers were observed but arrangementwas orderly (Figure 1(d))

32 Effects of Pain Threshold on Tail Flick Test The results ofthe pain threshold of tail flick test are shown in Figure 2 Thepain threshold of the control group at different time pointsdid not showobvious changesTheother three groups showedlower pain threshold from the second day after the operationAt 14 days the day before performing treatment comparedwith the control group the model EA and AK group had alower pain threshold (119875 lt 001 or119875 lt 005) and therewere nostatistical differences between themodel EA andAK groupsAfter treatments the pain threshold of the EA and AK grouprose Fromdays 17 to 28 comparedwith the control group thepain thresholds of the EA andAKgroupswere not statisticallydifferentHowever the pain threshold of themodel groupwassignificantly lower than the control group On days 21 and 28compared with the model group the pain threshold of theAK group rose significantly (119875 lt 005) From days 35 to 56there was no significant difference between all four groupsHowever Figure 1 shows a similar pain threshold between thecontrol EA and AK group which is higher than that of themodel group

33 Changes in IL-1120573 IL-10 TNF-120572 and TGF-120573 in thePeripheral Blood Fifty-six days after operation IL-1120573 andTNF-120572 levels in the model group were significantly higherthan that in the other three groups (119875 lt 005 or 119875 lt 001)Compared with the control group there was no significantdifference in IL-1120573 and TNF-120572 in the EA and AK groups(119875 gt 005) There was no significant difference in IL-1120573 andTNF-120572 between the EA and AK groups (119875 gt 005) (Figure 3)The IL-10 level in the model group was significantly higher

lowast

lowast

ControlModel

EAAK

8

6

4

2

0

Seru

m v

alue

of I

L-1120573

and

TNF-120572

(ng

mL)

IL-1120573 TNF-120572

Figure 3 Effects of AK on IL-1120573 and TNF-120572 in peripheral blood inthe following groups (119899 = 10 per group) control model EA andAK Compared with the control group lowast119875 lt 001 compared withthe model group 119875 lt 001 119875 lt 005

lowast

lowast

lowastlowastlowast

Seru

m v

alue

of I

L-10

and

TGF-120573

IL-10 (ngmL) TGF-120573 (pgmL)

350

300

250

200

150

100

50

0

ControlModel

EAAK

Figure 4 Effects of AK on IL-10 and TGF-120573 in peripheral bloodin the following groups (119899 = 10 per group) control modelEA and AK Compared with control group lowast119875 lt 001 lowastlowast119875 lt 005compared with model group 119875 lt 001

than that in the other three groups (119875 lt 001) Comparedwith the control group there was no significant differencein IL-10 in the EA and AK groups (119875 gt 005) There wereno significant differences in IL-10 among the EA and AKgroups (119875 gt 005) Compared with the control group TGF-120573 in the model EA and AK groups were significantly higher(119875 lt 001 or 119875 lt 005) Compared with the model groupTGF-120573 in the EA and AK groups were significantly lower(119875 lt 001) There was no difference between the EA and AKgroups (119875 gt 005 Figure 4)

Evidence-Based Complementary and Alternative Medicine 5

ControlModel

EAAK

lowast

lowast

lowastlowast

lowast300

270

240

210

180

150

120

90

60

30

0

Seru

m v

alue

of S

P an

d5

-HT

SP (pgmL) 5-HT (ngmL)

lowast

lowast

Figure 5 Effects of AK on SP and 5-HT in peripheral blood in thefollowing groups (119899 = 10 per group) control model EA and AKCompared with the control group lowast119875 lt 001 lowastlowast119875 lt 005 comparedwith themodel group 119875 lt 001 comparedwith the EA group 998771119875 lt005

34 Changes in SP and 5-Hydroxytryptamine in the PeripheralBlood Fifty-six days after operation compared with the con-trol group the model EA and AK groups had significantlyhigher SP levels in the peripheral blood (119875 lt 005 or 119875 lt001) Compared with the model group the EA and AKgroups had significantly lower SP levels in the peripheralblood (119875 lt 001) There was no significant difference inSP among the EA and AK groups (119875 gt 005) Comparedwith the control group the model EA and AK groups hadsignificantly higher 5-HT levels in the peripheral blood (119875 lt001) Compared with the model group the EA and AKgroups had significantly lower 5-HT levels in the peripheralblood (119875 lt 001) The 5-HT of the AK group was higher thanthat of the EA group (119875 lt 005 Figure 5)

4 Discussion

The term ldquotendon meridianrdquo in traditional Chinese medicineis part of the meridian system with functions of connectinglimbs and regulating the movement of joints Qi and bloodin meridians nourish some tissues involved in movementsuch as tendons ligaments fascia skeletal muscles andjoint capsules Some typical symptoms of tendon meridiandisease are pain numbness hypertonicity or stiffness ofmuscle or fascia even motor dysfunction which are possiblyequivalent to chronic injury of the motor system Thirdlumbar vertebrae transverse process syndrome is part of lowback pain according to TCM theory which is commonlybecause of qi and blood stagnation in the waist Disorderedblood circulation in the waist is caused by acute trauma achronic strain or invasion of cold pathogens The syndromeincludes spontaneous unilateral regional pain near the thirdlumbar vertebrae transverse process or referred pain tothe homolateral leg that is aggravated by bending down

with restricted range of waist motion and the presence ofhypersensitive nodules and local tenderness near the thirdlumbar vertebrae transverse process

After acute or chronic injury soft tissue adheres tothe tip of the third lumbar vertebrae transverse processwhich causes aseptic inflammation Inflammation causespathological changes such as edema exudation adhesion oreven continuous contracture in lesion tissues Nociceptiveterminals in muscle have a multitude of different receptors intheir membranes including matched receptors for moleculesthat are released from damaged tissue such as bradykininserotonin and prostaglandins [10] The continuous presenceof these inflammatorymediators and other chemicals may benecessary for persistent pain conditions [10]

Cytokine TNF-120572 is a mediator of chronic pain producedby the onset of inflammation [11] TNF-120572 can lead to NF-120581B-mediated transcription of IL-1 whichmediates inflammatoryhyperalgesia [12] IL-1120573 is one of the most important mem-bers of the interleukin-1 family When IL-1120573 binds to the IL-1 receptor the activation of IL-1 receptors induces changesin synaptic strength and results in behavioral hyperalgesia[13] Our results showed that serum TNF-120572 and IL-1120573 in themodel rats remained at a higher level than that in the normalrats This implies that the operation caused chronic inflam-mation The increased cytokines confirmed the activation ofimmune cells and the production of sterile inflammationThe rat model of embedding absorbent gelatin is suitablefor the observation of chronic inflammation and pain Afterelectroacupuncture and acupuncture knife treatments thelevels of serumTNF-120572 and IL-1120573were significantly lower thanthose in model rats Therefore both the electroacupunctureand acupuncture knife treatments can reduce the releaseof cytokines In addition the effect of anti-inflammation islong term Compared with EA rats there were no significantdifferences in AK rats in both serum levels of TNF-120572 andIL-1120573 However the contents of TNF-120572 and IL-1120573 in AKrats were closer to those in normal rats which suggeststhat acupuncture knife treatment is a more effective anti-inflammatory than electroacupuncture

IL-10 is a powerful anti-inflammatory cytokine that canreduce the production of proinflammatory cytokines suchas IL-1120573 IL-6 TNF-120572 and nerve growth factor IL-10-producing monocytes and macrophages promote the reso-lution of transient inflammatory hyperalgesia Impaired IL-10 production may represent a risk for developing chronicpain after inflammation [14] Moreover IL-10 gene therapyhas been proven useful in neuropathic pain rats [15] Ourresults showed that the serum IL-10 levels in model EA andAK rats were significantly higher than those in normal ratsHowever compared with model rats the serum levels in EArats and AK rats were significantly lower This result mayprove the therapeutic effect of downregulation of proinflam-matory cytokines by electroacupuncture and acupunctureknife treatments

TGF-120573 has been recognized as a growth factor withmultiple biological functions TGF-120573 plays an important rolein the regulation of immune cell function by limiting the pro-duction of inflammatory mediators including interleukin-2 and tumor necrosis factor [16] In chronic inflammation

6 Evidence-Based Complementary and Alternative Medicine

TGF-120573 is expressed and promotes wound healing and fibrosis[17] In addition TGF-120573 promotes the expression of endoge-nous opioids inhibiting the neuroimmune responses of glialcells and neurons in the spinal cord following peripheralinjuries Other undetermined mechanisms also contribute toits antiallodynic effect [18] Our results showed that the serumlevels of TGF-120573 in model EA and AK rats were significantlyhigher than those in normal rats which implies that theperipheral injury tissue is under the fibroplasia stage Thiswas also shown by pathological observations In addition thecontent of TGF-120573 in EA and AK rats is significantly lowerthan that in model rats This demonstrates that the level ofTGF-120573 declined because of the lighter inflammatory responsebrought by EA and AK treatments

Substance P (SP) is a neuropeptide released from nerveendings in many tissues which is directly related with painand plays an important role in inflammation [19] Peripheralinflammation and noxious stimuli may trigger the releaseof SP [20] thus lowering pain thresholds and resultingin allodynia [21] In the periphery administration of 5-HT can increase the excitability of myelinated (A120575 fibers)and unmyelinated afferents (C fibers) [22] Previous studyconfirmed that the inhibition of 5-HT secretion from mastcells can diminish the sensitization of sensory nerve terminals[23] Our results showed that 56 days after operation theconcentration of SP and 5-HT in the blood of EA and AKrats was lower than that in model rats and higher than thatin normal rats Therefore both EA and AK treatments con-tribute to relieving painThis may be because of alleviation oflocal inflammatory responses However compared with AKrats serum 5-HT of EA rats is significantly lowerThis impliesthat EA treatment is more effective for alleviating pain

5 Conclusions

In summary we demonstrated that acupuncture knife ther-apy is able to mitigate pain by controlling the inflammatoryresponse and is capable of restoring injury to soft tissueHowever the analgesic effect of electroacupuncture therapyis better than that of acupuncture knife therapy This maybe because of fewer treatment sessions for acupuncture knifetherapy (two times) than electroacupuncture therapy (sixtimes) accompanied by retaining the needle for 20minHowever acupuncture knife therapy has a better trendfor reducing inflammation In addition in spite of widercutting edge of acupuncture knife than acupuncture needlethe puncture depth of acupuncture knife is shallower thanacupuncture needle that is cutting in taut nodes and cordsof muscle or fascia located under skin without arteries veinsor nerves which can lead to irreversible damages The safetyof acupuncture knife is credible

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors show their deepest gratitude to ProfessorHanzhang Zhu for inventing the acupuncture knife treat-ment They thank him for his contribution to Chinesemedicine

References

[1] C M Ni A Zhao and J J He ldquoThe relationship betweenthe third lumbar transverse process syndrome and anatomyrdquoJournal of AnhuiMedical University vol 27 no 1 pp 11ndash13 1992

[2] C Q Guo S L Li J L Qiao et al ldquoMulticenter and randomizedcontrolled clinical trial of acupuncture knife therapy in treat-ment of third lumbar transverse process syndromerdquo Journal ofChengduUniversity of ChineseMedicine vol 35 no 1 pp 20ndash232012

[3] C Q Guo F H Dong S L Li et al ldquoEffects of acupotomy lysison local soft tissue tension in patients with the third lumbarvertebrae transverse process syndromerdquoChinese acupuncture ampMoxibustion vol 32 no 7 pp 617ndash620 2012

[4] B Hu L Liu C-Q Guo et al ldquoEffect of acupotomy lysis on SPand CCK-8 contents in hypothalamus and spinal cord in ratswith transverse process syndrome of the third lumbar vertebrardquoAcupuncture Research vol 33 no 1 pp 22ndash25 2008

[5] H M Sun B Hu X H Li et al ldquoEffect of acupuncture knifedissolution on pain threshold and ENK and 120573-EP in spinalcord and hypothalamus in rats with transverse syndrome of thethird lumbar vertebraerdquo Journal of Beijing University of ChineseMedicine vol 31 no 3 pp 176ndash179 2008

[6] E D de Avila R S de Molon D A de Godoi Goncalves andC M Camparis ldquoRelationship between levels of neuropeptideSubstance P in periodontal disease and chronic pain a literaturereviewrdquo Journal of Investigative and Clinical Dentistry vol 5 no2 pp 91ndash97 2014

[7] L Bardin ldquoThe complex role of serotonin and 5-HT receptorsin chronic painrdquo Behavioural Pharmacology vol 22 no 5-6 pp390ndash404 2011

[8] M de Goeij L T Van Eijk P Vanelderen et al ldquoSystemicinflammation decreases pain threshold in humans in vivordquoPLoS ONE vol 8 no 12 Article ID e84159 2013

[9] J RWang J L Qiao P Lu et al ldquoConstruct and research of ani-mal model for the third lumbar transverse process syndromerdquoChinese Journal of Physical Medicine and Rehabilitation vol 25no 7 pp 394ndash398 2003

[10] J P Shah J V Danoff M J Desai et al ldquoMochemicalsassociated with pain and inflammation are elevated in sites nearto and remote from active myofascial trigger pointsrdquo Archivesof Physical Medicine and Rehabilitation vol 89 no 1 pp 16ndash232008

[11] Q Liu L Jin B H Mahon M D Chordia F H Shen and XLi ldquoA novel treatment of neuroinflammation against low backpain by soluble fullerol nanoparticlesrdquo Spine vol 38 no 17 pp1443ndash1451 2013

[12] C A Dinarello A Simon and J W M van der Meer ldquoTreatinginflammation by blocking interleukin-1 in a broad spectrum ofdiseasesrdquoNature Reviews Drug Discovery vol 11 no 8 pp 633ndash652 2012

[13] K Ren and R Dubner ldquoInteractions between the immune andnervous systems in painrdquo Nature Medicine vol 16 no 11 pp1267ndash1276 2010

Evidence-Based Complementary and Alternative Medicine 7

[14] H L D M Willemen N Eijkelkamp A Garza Carbajal etal ldquoMonocytesmacrophages control resolution of transientinflammatory painrdquo Journal of Pain vol 15 no 5 pp 496ndash5062014

[15] E D Milligan E M Sloane S J Langer et al ldquoRepeatedintrathecal injections of plasmid DNA encoding interleukin-10produce prolonged reversal of neuropathic painrdquo Pain vol 126no 1ndash3 pp 294ndash308 2006

[16] N F Chen S Y Huang W F Chen et al ldquoTGF-1205731 attenuatesspinal neuroinflammation and the excitatory amino acid systemin rats with neuropathic painrdquo The Journal of Pain vol 14 no12 pp 1671ndash1685 2013

[17] Y Zhu T Colak M Shenoy et al ldquoTransforming growthfactor beta induces sensory neuronal hyperexcitability andcontributes to pancreatic pain and hyperalgesia in rats withchronic pancreatitisrdquoMolecular Pain vol 8 article 65 2012

[18] A Lantero M Tramullas A Diaz and M A Hurle ldquoTrans-forming growth factor-120573 in normal nociceptive processing andpathological pain modelsrdquoMolecular Neurobiology vol 45 no1 pp 76ndash86 2012

[19] M Munoz and R Covenas ldquoInvolvement of substance P andthe NK-1 receptor in human pathologyrdquo Amino Acids vol 46no 7 pp 1725ndash1750 2014

[20] B Bie and Z-Q Zhao ldquoPeripheral inflammation alters desen-sitization of substance P-evoked current in rat dorsal rootganglion neuronsrdquo European Journal of Pharmacology vol 670no 2-3 pp 495ndash499 2011

[21] K L Witt and J A Vilensky ldquoThe anatomy of osteoarthriticjoint painrdquo Clinical Anatomy vol 27 no 3 pp 451ndash454 2014

[22] L Bardin ldquoThe complex role of serotonin and 5-HT receptors inchronic painrdquo Behavior Pharmacollgy vol 22 no 5-6 pp 390ndash404 2011

[23] I Ferjan and M Lipnik-Stangelj ldquoChronic pain treatment Theinfluence of tricyclic antidepressants on serotonin release anduptake in mast cellsrdquo Mediators of Inflammation vol 2013Article ID 340473 7 pages 2013

Evidence-Based Complementary and Alternative Medicine 5

ControlModel

EAAK

lowast

lowast

lowastlowast

lowast300

270

240

210

180

150

120

90

60

30

0

Seru

m v

alue

of S

P an

d5

-HT

SP (pgmL) 5-HT (ngmL)

lowast

lowast

Figure 5 Effects of AK on SP and 5-HT in peripheral blood in thefollowing groups (119899 = 10 per group) control model EA and AKCompared with the control group lowast119875 lt 001 lowastlowast119875 lt 005 comparedwith themodel group 119875 lt 001 comparedwith the EA group 998771119875 lt005

34 Changes in SP and 5-Hydroxytryptamine in the PeripheralBlood Fifty-six days after operation compared with the con-trol group the model EA and AK groups had significantlyhigher SP levels in the peripheral blood (119875 lt 005 or 119875 lt001) Compared with the model group the EA and AKgroups had significantly lower SP levels in the peripheralblood (119875 lt 001) There was no significant difference inSP among the EA and AK groups (119875 gt 005) Comparedwith the control group the model EA and AK groups hadsignificantly higher 5-HT levels in the peripheral blood (119875 lt001) Compared with the model group the EA and AKgroups had significantly lower 5-HT levels in the peripheralblood (119875 lt 001) The 5-HT of the AK group was higher thanthat of the EA group (119875 lt 005 Figure 5)

4 Discussion

The term ldquotendon meridianrdquo in traditional Chinese medicineis part of the meridian system with functions of connectinglimbs and regulating the movement of joints Qi and bloodin meridians nourish some tissues involved in movementsuch as tendons ligaments fascia skeletal muscles andjoint capsules Some typical symptoms of tendon meridiandisease are pain numbness hypertonicity or stiffness ofmuscle or fascia even motor dysfunction which are possiblyequivalent to chronic injury of the motor system Thirdlumbar vertebrae transverse process syndrome is part of lowback pain according to TCM theory which is commonlybecause of qi and blood stagnation in the waist Disorderedblood circulation in the waist is caused by acute trauma achronic strain or invasion of cold pathogens The syndromeincludes spontaneous unilateral regional pain near the thirdlumbar vertebrae transverse process or referred pain tothe homolateral leg that is aggravated by bending down

with restricted range of waist motion and the presence ofhypersensitive nodules and local tenderness near the thirdlumbar vertebrae transverse process

After acute or chronic injury soft tissue adheres tothe tip of the third lumbar vertebrae transverse processwhich causes aseptic inflammation Inflammation causespathological changes such as edema exudation adhesion oreven continuous contracture in lesion tissues Nociceptiveterminals in muscle have a multitude of different receptors intheir membranes including matched receptors for moleculesthat are released from damaged tissue such as bradykininserotonin and prostaglandins [10] The continuous presenceof these inflammatorymediators and other chemicals may benecessary for persistent pain conditions [10]

Cytokine TNF-120572 is a mediator of chronic pain producedby the onset of inflammation [11] TNF-120572 can lead to NF-120581B-mediated transcription of IL-1 whichmediates inflammatoryhyperalgesia [12] IL-1120573 is one of the most important mem-bers of the interleukin-1 family When IL-1120573 binds to the IL-1 receptor the activation of IL-1 receptors induces changesin synaptic strength and results in behavioral hyperalgesia[13] Our results showed that serum TNF-120572 and IL-1120573 in themodel rats remained at a higher level than that in the normalrats This implies that the operation caused chronic inflam-mation The increased cytokines confirmed the activation ofimmune cells and the production of sterile inflammationThe rat model of embedding absorbent gelatin is suitablefor the observation of chronic inflammation and pain Afterelectroacupuncture and acupuncture knife treatments thelevels of serumTNF-120572 and IL-1120573were significantly lower thanthose in model rats Therefore both the electroacupunctureand acupuncture knife treatments can reduce the releaseof cytokines In addition the effect of anti-inflammation islong term Compared with EA rats there were no significantdifferences in AK rats in both serum levels of TNF-120572 andIL-1120573 However the contents of TNF-120572 and IL-1120573 in AKrats were closer to those in normal rats which suggeststhat acupuncture knife treatment is a more effective anti-inflammatory than electroacupuncture

IL-10 is a powerful anti-inflammatory cytokine that canreduce the production of proinflammatory cytokines suchas IL-1120573 IL-6 TNF-120572 and nerve growth factor IL-10-producing monocytes and macrophages promote the reso-lution of transient inflammatory hyperalgesia Impaired IL-10 production may represent a risk for developing chronicpain after inflammation [14] Moreover IL-10 gene therapyhas been proven useful in neuropathic pain rats [15] Ourresults showed that the serum IL-10 levels in model EA andAK rats were significantly higher than those in normal ratsHowever compared with model rats the serum levels in EArats and AK rats were significantly lower This result mayprove the therapeutic effect of downregulation of proinflam-matory cytokines by electroacupuncture and acupunctureknife treatments

TGF-120573 has been recognized as a growth factor withmultiple biological functions TGF-120573 plays an important rolein the regulation of immune cell function by limiting the pro-duction of inflammatory mediators including interleukin-2 and tumor necrosis factor [16] In chronic inflammation

6 Evidence-Based Complementary and Alternative Medicine

TGF-120573 is expressed and promotes wound healing and fibrosis[17] In addition TGF-120573 promotes the expression of endoge-nous opioids inhibiting the neuroimmune responses of glialcells and neurons in the spinal cord following peripheralinjuries Other undetermined mechanisms also contribute toits antiallodynic effect [18] Our results showed that the serumlevels of TGF-120573 in model EA and AK rats were significantlyhigher than those in normal rats which implies that theperipheral injury tissue is under the fibroplasia stage Thiswas also shown by pathological observations In addition thecontent of TGF-120573 in EA and AK rats is significantly lowerthan that in model rats This demonstrates that the level ofTGF-120573 declined because of the lighter inflammatory responsebrought by EA and AK treatments

Substance P (SP) is a neuropeptide released from nerveendings in many tissues which is directly related with painand plays an important role in inflammation [19] Peripheralinflammation and noxious stimuli may trigger the releaseof SP [20] thus lowering pain thresholds and resultingin allodynia [21] In the periphery administration of 5-HT can increase the excitability of myelinated (A120575 fibers)and unmyelinated afferents (C fibers) [22] Previous studyconfirmed that the inhibition of 5-HT secretion from mastcells can diminish the sensitization of sensory nerve terminals[23] Our results showed that 56 days after operation theconcentration of SP and 5-HT in the blood of EA and AKrats was lower than that in model rats and higher than thatin normal rats Therefore both EA and AK treatments con-tribute to relieving painThis may be because of alleviation oflocal inflammatory responses However compared with AKrats serum 5-HT of EA rats is significantly lowerThis impliesthat EA treatment is more effective for alleviating pain

5 Conclusions

In summary we demonstrated that acupuncture knife ther-apy is able to mitigate pain by controlling the inflammatoryresponse and is capable of restoring injury to soft tissueHowever the analgesic effect of electroacupuncture therapyis better than that of acupuncture knife therapy This maybe because of fewer treatment sessions for acupuncture knifetherapy (two times) than electroacupuncture therapy (sixtimes) accompanied by retaining the needle for 20minHowever acupuncture knife therapy has a better trendfor reducing inflammation In addition in spite of widercutting edge of acupuncture knife than acupuncture needlethe puncture depth of acupuncture knife is shallower thanacupuncture needle that is cutting in taut nodes and cordsof muscle or fascia located under skin without arteries veinsor nerves which can lead to irreversible damages The safetyof acupuncture knife is credible

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors show their deepest gratitude to ProfessorHanzhang Zhu for inventing the acupuncture knife treat-ment They thank him for his contribution to Chinesemedicine

References

[1] C M Ni A Zhao and J J He ldquoThe relationship betweenthe third lumbar transverse process syndrome and anatomyrdquoJournal of AnhuiMedical University vol 27 no 1 pp 11ndash13 1992

[2] C Q Guo S L Li J L Qiao et al ldquoMulticenter and randomizedcontrolled clinical trial of acupuncture knife therapy in treat-ment of third lumbar transverse process syndromerdquo Journal ofChengduUniversity of ChineseMedicine vol 35 no 1 pp 20ndash232012

[3] C Q Guo F H Dong S L Li et al ldquoEffects of acupotomy lysison local soft tissue tension in patients with the third lumbarvertebrae transverse process syndromerdquoChinese acupuncture ampMoxibustion vol 32 no 7 pp 617ndash620 2012

[4] B Hu L Liu C-Q Guo et al ldquoEffect of acupotomy lysis on SPand CCK-8 contents in hypothalamus and spinal cord in ratswith transverse process syndrome of the third lumbar vertebrardquoAcupuncture Research vol 33 no 1 pp 22ndash25 2008

[5] H M Sun B Hu X H Li et al ldquoEffect of acupuncture knifedissolution on pain threshold and ENK and 120573-EP in spinalcord and hypothalamus in rats with transverse syndrome of thethird lumbar vertebraerdquo Journal of Beijing University of ChineseMedicine vol 31 no 3 pp 176ndash179 2008

[6] E D de Avila R S de Molon D A de Godoi Goncalves andC M Camparis ldquoRelationship between levels of neuropeptideSubstance P in periodontal disease and chronic pain a literaturereviewrdquo Journal of Investigative and Clinical Dentistry vol 5 no2 pp 91ndash97 2014

[7] L Bardin ldquoThe complex role of serotonin and 5-HT receptorsin chronic painrdquo Behavioural Pharmacology vol 22 no 5-6 pp390ndash404 2011

[8] M de Goeij L T Van Eijk P Vanelderen et al ldquoSystemicinflammation decreases pain threshold in humans in vivordquoPLoS ONE vol 8 no 12 Article ID e84159 2013

[9] J RWang J L Qiao P Lu et al ldquoConstruct and research of ani-mal model for the third lumbar transverse process syndromerdquoChinese Journal of Physical Medicine and Rehabilitation vol 25no 7 pp 394ndash398 2003

[10] J P Shah J V Danoff M J Desai et al ldquoMochemicalsassociated with pain and inflammation are elevated in sites nearto and remote from active myofascial trigger pointsrdquo Archivesof Physical Medicine and Rehabilitation vol 89 no 1 pp 16ndash232008

[11] Q Liu L Jin B H Mahon M D Chordia F H Shen and XLi ldquoA novel treatment of neuroinflammation against low backpain by soluble fullerol nanoparticlesrdquo Spine vol 38 no 17 pp1443ndash1451 2013

[12] C A Dinarello A Simon and J W M van der Meer ldquoTreatinginflammation by blocking interleukin-1 in a broad spectrum ofdiseasesrdquoNature Reviews Drug Discovery vol 11 no 8 pp 633ndash652 2012

[13] K Ren and R Dubner ldquoInteractions between the immune andnervous systems in painrdquo Nature Medicine vol 16 no 11 pp1267ndash1276 2010

Evidence-Based Complementary and Alternative Medicine 7

[14] H L D M Willemen N Eijkelkamp A Garza Carbajal etal ldquoMonocytesmacrophages control resolution of transientinflammatory painrdquo Journal of Pain vol 15 no 5 pp 496ndash5062014

[15] E D Milligan E M Sloane S J Langer et al ldquoRepeatedintrathecal injections of plasmid DNA encoding interleukin-10produce prolonged reversal of neuropathic painrdquo Pain vol 126no 1ndash3 pp 294ndash308 2006

[16] N F Chen S Y Huang W F Chen et al ldquoTGF-1205731 attenuatesspinal neuroinflammation and the excitatory amino acid systemin rats with neuropathic painrdquo The Journal of Pain vol 14 no12 pp 1671ndash1685 2013

[17] Y Zhu T Colak M Shenoy et al ldquoTransforming growthfactor beta induces sensory neuronal hyperexcitability andcontributes to pancreatic pain and hyperalgesia in rats withchronic pancreatitisrdquoMolecular Pain vol 8 article 65 2012

[18] A Lantero M Tramullas A Diaz and M A Hurle ldquoTrans-forming growth factor-120573 in normal nociceptive processing andpathological pain modelsrdquoMolecular Neurobiology vol 45 no1 pp 76ndash86 2012

[19] M Munoz and R Covenas ldquoInvolvement of substance P andthe NK-1 receptor in human pathologyrdquo Amino Acids vol 46no 7 pp 1725ndash1750 2014

[20] B Bie and Z-Q Zhao ldquoPeripheral inflammation alters desen-sitization of substance P-evoked current in rat dorsal rootganglion neuronsrdquo European Journal of Pharmacology vol 670no 2-3 pp 495ndash499 2011

[21] K L Witt and J A Vilensky ldquoThe anatomy of osteoarthriticjoint painrdquo Clinical Anatomy vol 27 no 3 pp 451ndash454 2014

[22] L Bardin ldquoThe complex role of serotonin and 5-HT receptors inchronic painrdquo Behavior Pharmacollgy vol 22 no 5-6 pp 390ndash404 2011

[23] I Ferjan and M Lipnik-Stangelj ldquoChronic pain treatment Theinfluence of tricyclic antidepressants on serotonin release anduptake in mast cellsrdquo Mediators of Inflammation vol 2013Article ID 340473 7 pages 2013

6 Evidence-Based Complementary and Alternative Medicine

TGF-120573 is expressed and promotes wound healing and fibrosis[17] In addition TGF-120573 promotes the expression of endoge-nous opioids inhibiting the neuroimmune responses of glialcells and neurons in the spinal cord following peripheralinjuries Other undetermined mechanisms also contribute toits antiallodynic effect [18] Our results showed that the serumlevels of TGF-120573 in model EA and AK rats were significantlyhigher than those in normal rats which implies that theperipheral injury tissue is under the fibroplasia stage Thiswas also shown by pathological observations In addition thecontent of TGF-120573 in EA and AK rats is significantly lowerthan that in model rats This demonstrates that the level ofTGF-120573 declined because of the lighter inflammatory responsebrought by EA and AK treatments

Substance P (SP) is a neuropeptide released from nerveendings in many tissues which is directly related with painand plays an important role in inflammation [19] Peripheralinflammation and noxious stimuli may trigger the releaseof SP [20] thus lowering pain thresholds and resultingin allodynia [21] In the periphery administration of 5-HT can increase the excitability of myelinated (A120575 fibers)and unmyelinated afferents (C fibers) [22] Previous studyconfirmed that the inhibition of 5-HT secretion from mastcells can diminish the sensitization of sensory nerve terminals[23] Our results showed that 56 days after operation theconcentration of SP and 5-HT in the blood of EA and AKrats was lower than that in model rats and higher than thatin normal rats Therefore both EA and AK treatments con-tribute to relieving painThis may be because of alleviation oflocal inflammatory responses However compared with AKrats serum 5-HT of EA rats is significantly lowerThis impliesthat EA treatment is more effective for alleviating pain

5 Conclusions

In summary we demonstrated that acupuncture knife ther-apy is able to mitigate pain by controlling the inflammatoryresponse and is capable of restoring injury to soft tissueHowever the analgesic effect of electroacupuncture therapyis better than that of acupuncture knife therapy This maybe because of fewer treatment sessions for acupuncture knifetherapy (two times) than electroacupuncture therapy (sixtimes) accompanied by retaining the needle for 20minHowever acupuncture knife therapy has a better trendfor reducing inflammation In addition in spite of widercutting edge of acupuncture knife than acupuncture needlethe puncture depth of acupuncture knife is shallower thanacupuncture needle that is cutting in taut nodes and cordsof muscle or fascia located under skin without arteries veinsor nerves which can lead to irreversible damages The safetyof acupuncture knife is credible

Conflict of Interests

The authors declare that there is no conflict of interestsregarding the publication of this paper

Acknowledgment

The authors show their deepest gratitude to ProfessorHanzhang Zhu for inventing the acupuncture knife treat-ment They thank him for his contribution to Chinesemedicine

References

[1] C M Ni A Zhao and J J He ldquoThe relationship betweenthe third lumbar transverse process syndrome and anatomyrdquoJournal of AnhuiMedical University vol 27 no 1 pp 11ndash13 1992

[2] C Q Guo S L Li J L Qiao et al ldquoMulticenter and randomizedcontrolled clinical trial of acupuncture knife therapy in treat-ment of third lumbar transverse process syndromerdquo Journal ofChengduUniversity of ChineseMedicine vol 35 no 1 pp 20ndash232012

[3] C Q Guo F H Dong S L Li et al ldquoEffects of acupotomy lysison local soft tissue tension in patients with the third lumbarvertebrae transverse process syndromerdquoChinese acupuncture ampMoxibustion vol 32 no 7 pp 617ndash620 2012

[4] B Hu L Liu C-Q Guo et al ldquoEffect of acupotomy lysis on SPand CCK-8 contents in hypothalamus and spinal cord in ratswith transverse process syndrome of the third lumbar vertebrardquoAcupuncture Research vol 33 no 1 pp 22ndash25 2008

[5] H M Sun B Hu X H Li et al ldquoEffect of acupuncture knifedissolution on pain threshold and ENK and 120573-EP in spinalcord and hypothalamus in rats with transverse syndrome of thethird lumbar vertebraerdquo Journal of Beijing University of ChineseMedicine vol 31 no 3 pp 176ndash179 2008

[6] E D de Avila R S de Molon D A de Godoi Goncalves andC M Camparis ldquoRelationship between levels of neuropeptideSubstance P in periodontal disease and chronic pain a literaturereviewrdquo Journal of Investigative and Clinical Dentistry vol 5 no2 pp 91ndash97 2014

[7] L Bardin ldquoThe complex role of serotonin and 5-HT receptorsin chronic painrdquo Behavioural Pharmacology vol 22 no 5-6 pp390ndash404 2011

[8] M de Goeij L T Van Eijk P Vanelderen et al ldquoSystemicinflammation decreases pain threshold in humans in vivordquoPLoS ONE vol 8 no 12 Article ID e84159 2013

[9] J RWang J L Qiao P Lu et al ldquoConstruct and research of ani-mal model for the third lumbar transverse process syndromerdquoChinese Journal of Physical Medicine and Rehabilitation vol 25no 7 pp 394ndash398 2003

[10] J P Shah J V Danoff M J Desai et al ldquoMochemicalsassociated with pain and inflammation are elevated in sites nearto and remote from active myofascial trigger pointsrdquo Archivesof Physical Medicine and Rehabilitation vol 89 no 1 pp 16ndash232008

[11] Q Liu L Jin B H Mahon M D Chordia F H Shen and XLi ldquoA novel treatment of neuroinflammation against low backpain by soluble fullerol nanoparticlesrdquo Spine vol 38 no 17 pp1443ndash1451 2013

[12] C A Dinarello A Simon and J W M van der Meer ldquoTreatinginflammation by blocking interleukin-1 in a broad spectrum ofdiseasesrdquoNature Reviews Drug Discovery vol 11 no 8 pp 633ndash652 2012

[13] K Ren and R Dubner ldquoInteractions between the immune andnervous systems in painrdquo Nature Medicine vol 16 no 11 pp1267ndash1276 2010

Evidence-Based Complementary and Alternative Medicine 7

[14] H L D M Willemen N Eijkelkamp A Garza Carbajal etal ldquoMonocytesmacrophages control resolution of transientinflammatory painrdquo Journal of Pain vol 15 no 5 pp 496ndash5062014

[15] E D Milligan E M Sloane S J Langer et al ldquoRepeatedintrathecal injections of plasmid DNA encoding interleukin-10produce prolonged reversal of neuropathic painrdquo Pain vol 126no 1ndash3 pp 294ndash308 2006

[16] N F Chen S Y Huang W F Chen et al ldquoTGF-1205731 attenuatesspinal neuroinflammation and the excitatory amino acid systemin rats with neuropathic painrdquo The Journal of Pain vol 14 no12 pp 1671ndash1685 2013

[17] Y Zhu T Colak M Shenoy et al ldquoTransforming growthfactor beta induces sensory neuronal hyperexcitability andcontributes to pancreatic pain and hyperalgesia in rats withchronic pancreatitisrdquoMolecular Pain vol 8 article 65 2012

[18] A Lantero M Tramullas A Diaz and M A Hurle ldquoTrans-forming growth factor-120573 in normal nociceptive processing andpathological pain modelsrdquoMolecular Neurobiology vol 45 no1 pp 76ndash86 2012

[19] M Munoz and R Covenas ldquoInvolvement of substance P andthe NK-1 receptor in human pathologyrdquo Amino Acids vol 46no 7 pp 1725ndash1750 2014

[20] B Bie and Z-Q Zhao ldquoPeripheral inflammation alters desen-sitization of substance P-evoked current in rat dorsal rootganglion neuronsrdquo European Journal of Pharmacology vol 670no 2-3 pp 495ndash499 2011

[21] K L Witt and J A Vilensky ldquoThe anatomy of osteoarthriticjoint painrdquo Clinical Anatomy vol 27 no 3 pp 451ndash454 2014

[22] L Bardin ldquoThe complex role of serotonin and 5-HT receptors inchronic painrdquo Behavior Pharmacollgy vol 22 no 5-6 pp 390ndash404 2011

[23] I Ferjan and M Lipnik-Stangelj ldquoChronic pain treatment Theinfluence of tricyclic antidepressants on serotonin release anduptake in mast cellsrdquo Mediators of Inflammation vol 2013Article ID 340473 7 pages 2013

Evidence-Based Complementary and Alternative Medicine 7

[14] H L D M Willemen N Eijkelkamp A Garza Carbajal etal ldquoMonocytesmacrophages control resolution of transientinflammatory painrdquo Journal of Pain vol 15 no 5 pp 496ndash5062014

[15] E D Milligan E M Sloane S J Langer et al ldquoRepeatedintrathecal injections of plasmid DNA encoding interleukin-10produce prolonged reversal of neuropathic painrdquo Pain vol 126no 1ndash3 pp 294ndash308 2006

[16] N F Chen S Y Huang W F Chen et al ldquoTGF-1205731 attenuatesspinal neuroinflammation and the excitatory amino acid systemin rats with neuropathic painrdquo The Journal of Pain vol 14 no12 pp 1671ndash1685 2013

[17] Y Zhu T Colak M Shenoy et al ldquoTransforming growthfactor beta induces sensory neuronal hyperexcitability andcontributes to pancreatic pain and hyperalgesia in rats withchronic pancreatitisrdquoMolecular Pain vol 8 article 65 2012

[18] A Lantero M Tramullas A Diaz and M A Hurle ldquoTrans-forming growth factor-120573 in normal nociceptive processing andpathological pain modelsrdquoMolecular Neurobiology vol 45 no1 pp 76ndash86 2012

[19] M Munoz and R Covenas ldquoInvolvement of substance P andthe NK-1 receptor in human pathologyrdquo Amino Acids vol 46no 7 pp 1725ndash1750 2014

[20] B Bie and Z-Q Zhao ldquoPeripheral inflammation alters desen-sitization of substance P-evoked current in rat dorsal rootganglion neuronsrdquo European Journal of Pharmacology vol 670no 2-3 pp 495ndash499 2011

[21] K L Witt and J A Vilensky ldquoThe anatomy of osteoarthriticjoint painrdquo Clinical Anatomy vol 27 no 3 pp 451ndash454 2014

[22] L Bardin ldquoThe complex role of serotonin and 5-HT receptors inchronic painrdquo Behavior Pharmacollgy vol 22 no 5-6 pp 390ndash404 2011

[23] I Ferjan and M Lipnik-Stangelj ldquoChronic pain treatment Theinfluence of tricyclic antidepressants on serotonin release anduptake in mast cellsrdquo Mediators of Inflammation vol 2013Article ID 340473 7 pages 2013