Research on Effects of the Thermal Stimulation by Moxibustion at

Hindawi Publishing CorporationEvidence-Based Complementary and Alternative MedicineVolume 2013 Article ID 545707 7 pageshttpdxdoiorg1011552013545707

Research ArticleResearch on Effects of the Thermal Stimulation byMoxibustion at Different Temperatures on Cardiac Function inRats and on Mast Cells in the Local Site of Moxibustion

Yao-Shuai Wang Jian-Bin Zhang Jin-Feng Jiang and Ling-Ling Wang

The Second Clinical College Nanjing University of Chinese Medicine Nanjing 210023 China

Correspondence should be addressed to Ling-Ling Wang wanglinglgmailcom

Received 1 March 2013 Revised 20 June 2013 Accepted 21 June 2013

Academic Editor Yi-Hung Chen

Copyright copy 2013 Yao-Shuai Wang et alThis is an open access article distributed under theCreativeCommonsAttribution Licensewhich permits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Objective To observe effects of the thermal stimulation bymoxibustion at different temperatures on cardiac function in brachycardiarat model and on mast cells in the local site of moxibustion at the Ximen Acupoint and to compare the differences of the effectsof moxibustion at different temperaturesMethod Establish the brachycardia rat model with propranolol and observe effects of thethermal stimulation by moxibustion at different temperatures (38∘C and 46∘C) Results The thermal stimulation by moxibustion at2 temperatures may increase HR MAP LVSP and +dpdtmax and reduce t-dpdtmax in brachycardia rats the 46∘C moxibustiongroup shows greater regulating effects on cardiac function in rats than that in the 38∘Cmoxibustion group (119875 lt 005) The thermalstimulation bymoxibustion at 2 temperaturesmay promote degranulation ofmast cells in the local site ofmoxibustion at the XimenAcupoint the degranulation rate in the 46∘C moxibustion group is higher than that in the 38∘C moxibustion group (119875 lt 005)Conclusion There is a certain association between the effect on the target organ and the effect in the local site of moxibustion Themoxibustion effect possibly resulted from local mast cells degranulation and different thermoreceptors activated by the thermalstimulation at different temperatures

1 Introduction

As an importantmethod in the traditional Chinesemedicinemoxibustion is featured mainly with the ldquowarmrdquo advantageby which it carries out stimulation at the Acupoint or thelesions and works by ldquostimulation from the outside andregulation from the insiderdquo The skin is the direct actionsite of moxibustion stimulation selection of appropriatewarm thermal stimulation to local skin characteristics ofresponses to stimulation and activation of relevant regulatingmechanisms are based on the biological characteristics ofskin that is to say to explore the effect law and actionmechanism of moxibustion attention has to be paid to theaction characteristics of moxibustion and the characteristicsof the action object of moxibustion Modern neurobiologicalstudies have demonstrated that [1] the body and skin tissuesare distributed with protein receptors (transient receptorpotential (TRP)) sensing different temperature ranges whichhave their respective thresholds in sensing temperatures and

may be activated by different warm thermal stimulation Thecharacteristics in the change of local temperatures caused bymoxibustion therapy are greatly consistent with categoriesand distribution characteristics of relevant temperature-sensing subtypes of TRP such as the TRPV1 in the TRP familywhich may be activated by thermal stimulation (gt43∘C) [2]Recent studies have found that TRPV1 expression and activa-tion of skin cutin cells may induce inflammatory reaction [3]Activated TRPV1 can make mast cell degranulation releaseproinflammatory and itch causing media to participate inthe inflammatory reaction [4] The inflammatory reactionis associated with pathological physiological function in thecardiac arrhythmias [5] Moreover TRPV1 has an importantregulating effect on the cardiovascular system [6] Moxibus-tion can effectively regulate blood blood flow and microcir-culation condition and diastolic function [7 8] Studies haveshown effects of acu-moxibustion associated with mast cellsin acupoints [9] From the previously mentioned attentionneeds to be paid to what effects may be produced by the

2 Evidence-Based Complementary and Alternative Medicine

thermal stimulation at two temperatures below and above43∘C on cardiac function in rates as well as on skin in thelocal site of moxibustion whether there is any differenceor association of effects and whether they may provideany basis for optimizing the clinical operating program ofmoxibustion

2 Materials and Methods

21 Materials

211 Animals and Grouping Healthy male SD rats of cleangrade with body weight of 280ndash380 g Certificate no SCXK(Shanghai) 2007-0005 were purchased from Shanghai SlacLaboratory Animal Co Ltd and bred in batches Theexperiment was carried out in 1 week after the rats wereaccommodated The rats were divided into the followinggroups by the random number table Model Control GroupMoxibustionGroup 1 andMoxibustionGroup 2Thenumberof rats in each group would support to obtain 8 complete datasuccessfully During the experiment the animal treatmentmeasures were in strict accordance with the regulationsin the Instructive Opinions on Humanitarian Treatment ofLaboratory Animals issued in 2006 by theMinistry of Scienceand Technology of the Peoplersquos Republic of China and theprovisions for care and use of laboratory animals by NanjingUniversity of Traditional Chinese Medicine

212 Main Laboratory Equipment Moxa strips for animals(in the diameter of 7mm Nanyang Chinese Medicine MoxaCompany) thermostatic operating table for animals (ST-1model Chengdu Instrument Factory) multichannel physio-logical signal acquisition system (RM 6240 model ChengduInstrument Factory) temperature measuring device (HC-04 model Hangzhou Hongchang Technology Company)microsyringe pump (ALC-IP800 Shanghai Alcott BiotechCo Ltd) full automatic tissue processor (TP1020 GermanLEICA Company) paraffin slicing machine (RM2145 Ger-man LEICA Company) optical microscope (BX60 JapaneseOLYMPUS Company) and video camera (DP71 ImageAcquisition System Japanese OLYMPUS Company) wereused

22 Methods

221 Electrophysiological Experimental Method Fast a maleSD rat overnight for 12 hours with free access to water weighthe rat carry out anesthesia of the rat with 20 urethane(ethyl carbamate) in the dose of 1 gkg body weight fix the ratin the thermostatic operating table for animals and connectwith the electrocardiograph and the temperature transducer(via the anus of the rat) once the electrocardiogram andbody temperature of the rat are stable (37∘C) carry outintubation in the left ventricle as well as in the femoral arteryand femoral vein of the right lower limb to measure leftventricular pressure and arterial pressure and administer thedrug intravenously Model control group 10 minutes afterthe operation if the measurement indexes are stable start

recording data 5 minutes later rapid inject 04 inderal(4mgkg) via the femoral vein then slowly inject intra-venously to maintain inderal at the dose of 025mgkgminwith the microsyringe pump for 60min to establish thebradycardia model no any other interventions are carriedout Moxibustion group 1 establish the model as describedin the model control group At 20min during administrationof inderal at the maintenance dose ignite the customizedslender moxa strip (20 cm long and in the diameter of7mm) carry out moxibustion directly to stimulate at theXimen(PC4) Acupoint in the left upper limb of the rat for10min place the probe of the temperature controller atthe surface of the skin at the acupoint for moxibustion tomonitor real-time temperature and control the temperaturesin the local skin for moxibustion at 38∘C plusmn 1∘C (Figure 1(a))the flaming moxa strip and skin are 35 plusmn 5mm apartafter moxibustion continue observation for another 30minMoxibustion group 2 establish the model as described inthe model control group At 20min during administrationof inderal in the maintenance dose carry out moxibustionfor thermal stimulation of 10min at 46∘C carry out theintervention similar to that as described in moxibustiongroup 1 during thermal stimulation by moxibustion controlthe temperature in the local skin for moxibustion at 46∘C plusmn1∘C Figure 1(b) and the flaming moxa strip and skin are 12 plusmn2mm apart after moxibustion continue observation foranother 30min During moxibustion gently tap any moxaashes at the tip of the moxa strip to the ashtray based onthe combustion state of the moxa strip to always maintaincleanliness of the operating interface The Ximen Acupointin the left limb is selected the positioning method of theacupoint is the anthropomorphic comparative method thatis Ximen In the inner side at the 12 folding point of theforearm and in the middle of the gap between the radius andthe ulna The reason for the choice of the temperature is thatbased on our observations less than 38∘C thermal stimulusthe human body for the warm feeling is not obvious andabove 46∘C thermal stimulus the human body for the hotfeeling is very obvious difficult to continue to endure Duringthe whole course using RM6240B multichannel physiologyrecorder (parameters acquisition frequency 1 kz scan speed40msdiv and filter frequency 30Hz) to simultaneouslymonitor and record many indexes for cardiac function andhemodynamics including heart rate (min) mean femoralartery pressure (mmHg) left ventricular systolic pressure(mmHg) maximum rate of left ventricular pressure rise(mmHgs) and time (ms) from the beginning of left ventric-ular systole to the maximum rate of left ventricular pressurerate and so forth

222 Morphological Experimental Method

(1) Obtain Materials After completion of the monitoring andrecording of the electrophysiology experiment sacrifice therats and obtain materials Rapidly extract skin tissues in thelocal site at the XimenAcupoint in the rats of each groupwiththe ophthalmic forceps and the surgical scissors and placeinto 10 formaldehyde solution to store and fix for 1 week

Evidence-Based Complementary and Alternative Medicine 3

(a) (b)

Figure 1 Sample curve picture of the point skin temperature during hot stimulation process of moxibustion group Moxibustion Group 1 (a)Moxibustion Group 2 (b)

(2) Processing prior to Staining (1) Trimming take out theskin tissues that are sheared and then place them in 10formaldehyde solution and fix them for 1 week trim thetissues with the surgical knife to a size of approximately07 cm lowast 07 cm lowast 03 cm and then place them into 70alcohol to remove any residual formaldehyde solution in thetissues (2) Dehydration place the trimmed tissues into thefull automatic tissue processor for dehydration (3) Embed-ding carry out paraffin embedding of the processed tissues(4) Slicing carry out common slicing by the slicing machineto obtain slices in the thickness of 5-6120583m and place theminto hot water to be flattened Then slowly take out the slicesrapidly shake off water and then slowly place them on thewell-labeled glass slides to prevent any air bubbles formedbetween tissue and slide in case of any air bubbles prick offcarefully with the acupuncture needle Bake on a slice-bakingplate until the paraffin turns transparent and place into theslice-baking oven bake overnight at 60ndash64∘C and then allowto cool for use

(3) Toluidine Blue Staining (1) Commonly remove paraffinfrom slices and then wash with water (2) dip-dye with05 toluidine blue solution (to 05 g of toluidine blue dyeadd water to the volume of 100mL) (3) wash with waterto remove any excessive staining solution (4) carry outdifferentiation with the differentiation solution (mix 05mLof glacial acetic acid and 995mL of distilled water andshake well) (5) wash with water and air-dry (6) carry outdehydration gradiently with alcohol (7) treat with xyleneuntil transparent and (8) carry out fixation of slices withneutral balsam

223 Statistical Method The data are expressed as mean plusmnstandard deviation (119883plusmn119878) and are analyzed and processed bythe SPSS170 statistical softwareThe data obtained from eachgroup are subject first to the homogeneity test of variance In

case of homogeneity of variance the one-factor analysis ofvariance is used if there is difference of population meansthe pair-wise comparisons are carried out by the LSD test (incase of heterogeneity of variance the pair-wise comparisonsare carried out by Brown-Forsythe approximate analysis ofvariance and Games-Howell method) 119905 test is used forcomparison between two independent samples It is regardedas significant in case of 119875 lt 005

3 Results

During the experiment all deaths due to anesthesia operativeaccidents or drug administration and so forth as well as allabnormal responses to intervention of the established modelare not included for observation 8 valid experimental caseswere obtained in each group so a total of 24 cases wereobtained

31 Results from Electrophysiological Experiment Statisticalanalysis was carried out by the 5min means for each ofthe indexes collected such as HR MAP and cardiac func-tion (LVSP +dpdtmax and t-dpdtmax) To reduce anystatistical errors due to individual differences the changesbefore and after intervention in each experimental animalwere expressed in percent Change Rate () = (value afterintervention minus value before intervention)value before inter-vention times 100 The results were provided as follows

311 Characteristics in the Changes of HR and MAP in theBrachycardia Rat Models Induced by Inderal The admin-istration of inderal resulted in rapid decrease of HR andMAP in rats (on average in 60 minutes HR was decreasedby 3498 plusmn 586 and MAP was decreased by 3321 plusmn563) In 20 minutes after administration of inderal themodel was stable successfully during 40-min maintenance


Table 1Mean change percentage of themodel groupHRMAP ()

Model group 119873 HR MAP60 minutes after administrationof inderal 8 3498 plusmn 586 3321 plusmn 563

After 20-min administrationof inderal 8 minus016 plusmn 198 037 plusmn 209

after 20min administration of inderal the mean change rateof HR was minus016 plusmn 198 and that of MAP was 037 plusmn209 The results showed that the method was able toestablish brachycardia rat models with stable effects (Table 1)

312 Waveform Changes of Left Ventricular Pressure andArterial Pressure before and after Model Establishment andIntervention byMoxibustion After themodel was establishedsuccessfully with inderal the waveforms of left ventricularpressure and arterial pressure were obviously decreased thannormal after thermal stimulation bymoxibustion at 38∘Cand46∘C the waveforms of left ventricular pressure and arterialpressure were recovered to some extents than those in themodel (Figure 2)

313 Intergroup Comparisons of Mean Change Rates of Eachof the Indexes Such As HR MAP LVSP +dpdtmax and t-dpdtmax after Intervention byMoxibustion Comparedwiththose in the model control group the thermal stimulation intheMoxibustion group 1 was able to increaseHRMAP LVSPand +dpdtmax and reduce t-dpdtmax in rats however theeffects were not significant (119875 gt 005) Compared with thosein the model control group the thermal stimulation in theMoxibustion group 2 was able to increase HR MAP LVSPand +dpdtmax and reduce t-dpdtmax in rats and the effectswere significant (119875 lt 005) moreover the effects of thethermal stimulation in the moxibustion group 2 were higherthan those in the moxibustion group 1 (119875 lt 005) (Table 2)

32 Results from the Morphological Experiment The mor-phology of mast cells was observed under the 10 times 40 opticalmicroscope and mast cells with degranulation in skin werecounted

321 Results of Morphological Observations of Mast CellsFindings of mast cells in slices of skin tissues in the area of theXimen Acupoint were observed under optical microscope(1) Model control group the mast cells were of smoothcell walls and of complete morphology (Figure 3(a)) (2)moxibustion group 1 the mast cells in the local site wereof incomplete morphology and were with ruptured cellmembranes (Figure 3(b)) (3) moxibustion group 2 the mastcells in the local site were with ruptured cell membranes anda large amount of granules were scattered around the cellbody and there were significant degranulations (Figure 3(c))

322 Degranulation Rate of Mast Cells The criteria forcounting mast cells with degranulation were count as 1 whenthe mast cell was with ruptured cell wall and there were

scattered granules in the matrix Degranulation rate = countof mast cells with degranulationtotal count of mast cells times100 The mean of degranulation rates from 4 slices whichwere closest to the local site of moxibustion was calculatedin each tissue sample add the mean of degranulation ratesof this group all tissue samples together and divided by thenumber of samples in each group then the degranulation rateof mast cells could be calculated

If no interventionwas carried out to the skin at the XimenAcupoint in rates the degranulation rate in the local site wasas lowas 129 after thermal stimulation bymoxibustion thedegranulation rate of mast cells was significantly increased(119875 lt 005) (Table 3)

4 Discussion

From ancient times moxibustion is one of the importantmethods in treatment of cardiovascular diseases in clinicalpractices It is a crucial problem for moxibustion researchhow to objectively assess the effects of moxibustion on heartand blood vessels to explore the possible mechanism fordifferences of effects resulted from stimulation by moxibus-tion at different temperatures as an influencing factor and toprovide a basis for optimizing the clinical operating programofmoxibustion In this experiment propranolol a120573-receptorblocker was used to establish the acute brachycardia animalmodel [10 11] It is an easy and simple technique with goodrepeatability The narcotic agent urethane is of wide safetymargin with an easily controlled dose and has small toxicityto heartThe cardiovascular rat model is of good stability andmoxibustion at the Ximen Acupoint has good effects againstarrhythmia they have been verified well in the previousstudies of the research group [12 13] In this experimentthe indexes including HR MAP LVSP +dpdtmax and t-dpdtmax were analyzed comprehensively in this way itmay completely reflect the effects of the thermal stimulationby moxibustion on myocardial contractility and peripheralcirculation center function HR may reflect change in themyocardial contractility visually and clearly After opening ofthe aortic valve the left ventricle ejects blood to the arteryThe stronger the left ventricle contracts the more blood itejectss and then the higher the blood pressure is the afterloadresisting ejection of the left ventricle is mainly the peripheralblood pressure (the mean femoral artery pressure was mainlyobserved in this experiment) The left ventricular systolicfunction is associated with the afterload of the left ventricleMAPmay reflect the change of the peripheral circulation cen-ter function +dpdtmax is very sensitive to interventions byvarious variable stresses and to a certain extent is influencedby and positively correlated with heart rate and the preloadand afterload the increase of +dpdtmax with concurrentreduction of t-dpdtmax suggests increase of myocardialcontractility The results of this study found that the thermalstimulation by moxibustion at 2 temperatures (38∘C and46∘C)may promote increase of HR andMAP in propranolol-induced brachycardia model rats recovery of LVSP and+dpdtmax and reduction of t-dpdtmax suggesting that thethermal stimulation by moxibustion at 2 temperatures (38∘C


(a) (b)

(c) (d)

Figure 2 Sample waveshape picture of the LVP AP of each group normal condition (a) the model was established successfully with inderal(b) after thermal stimulation at 38∘C (c) after thermal stimulation at 46∘C (d)

Table 2 Mean change percentage of the HR MAP LVSP +dpdtmax and t-dpdtmax of different groups after moxibustion treatment

Group 119873 HR (min) MAP (mmHg) LVSP (mmHg) +dpdtmax (mmHgs) t-dpdtmax (ms)Model control group 8 minus0023 plusmn 0099 minus0033 plusmn 0086 0045 plusmn 0093 0057 plusmn 014 0018 plusmn 0113

Moxibustion group 1 8 0044 plusmn 0212 0063 plusmn 0078 0087 plusmn 003 014 plusmn 0052 minus0026 plusmn 004

Moxibustion group 2 8 0143 plusmn 0074lowast998771

0425 plusmn 0284lowast998771



minus019 plusmn 016lowast998771

Analysis of variance119865 10660 14963 13875 13568 7217119875 0001 0000 0000 0000 0004

Note the values in the table for each index value variation rateCompared with model control group lowast119875 lt 005Compared with moxibustion group 1 998771119875 lt 005

and 46∘C) has the effects of improving cardiac function inbrachycardia rats and the effects of the thermal stimulationby moxibustion at 46∘C are obviously stronger than those ofthe thermal stimulation by moxibustion at 38∘C Our resultsare in conformity with ancient doctorsrsquo point of view thatenough strength of thermal stimulation achieve good curativeeffect

The ancient doctors emphasis moxibustion heat stimu-lates the skin local inflammatory response is beneficial tothe treatment of diseases Our study demonstrated that thethermal stimulation by moxibustion may influence the mor-phology ofmast cells in the local site ofmoxibustion promotetheir degranulation and similarly the thermal stimulation bymoxibustion at 46∘C is obviously stronger than that by mox-ibustion at 38∘C The results suggested that higher or lowermast cell degranulation rate influenced the improvementof cardiac function so the characteristics of warm thermalstimulation by moxibustion are certainly associated with theeffects on the target organ as well as on the local site of

moxibustionThe local site ofmoxibustion is the direct objectofmoxibustion action and is one of the key links for the start-ing mechanism of the moxibustion effects The therapeuticeffect of moxibustion is to regulate the body by stimulatingthe acupoints while the regulating effects are dependent onreceptors in the acupoint area Recent studies have believedthat the skin is an important neuroendocrine organ whichmay synthesize and secrete many neurotransmitters andpeptide substances [14] while the TRP channel is probablythe significant potentialmolecular target of thewarm thermaleffects bymoxibustionTheTRP channel is widely distributedin mast cells in the body [15] mainly including TRPV1 andTRPV2 which opening effects may directly influence thefunction and efficacy ofmast cells Especially TRPV1 calciumion channels of mast cells may participate in regulatingmast cell biological functions and the pathological process[16] Some scholars proposed the ldquoaxonal reflex-linkagehypothesisrdquo [17] in which the synaptic connection betweenmast cells and nerve fiber acts as a relay baton and the


(a) (b)

(c)

Figure 3 Morpohologies of Mast cells in the Local Site of the Ximen Acupoint Model Control Group (a) Moxibustion Group 1 (b)Moxibustion Group 2 (c)

Table 3 Degranulation rate of mast cells

Group 119873 Degranulation rate of mast cells Model control group 8 129 plusmn 28

Moxibustion group 1 8 376 plusmn 59lowast


Note compared with model control group lowast119875 lt 005compared with moxibustion group 1 998771119875 lt 005

information is transmitted in the form of stimulating mastcells to degranulate and release active substances and then toactivate adjacent nerve terminals Galli proposed ldquoMast cellscytokine cascade reactionrdquo [18] based on the characteristicsof Mast cells secreting many transmitters and cytokinesTherefore we deduced that different moxibustion methodsmay produce different characteristics of temperature changesactivate different TRP subtypes induce different biologicaleffects and finally achieve different therapeutic effects

In this study the relevant new biological knowledge ofthe skin was used as the basis for this study to focus ondifferences of the effects due to warm thermal stimulationby moxibustion at different temperatures and to exploretheir association with physiological feedback responses fromdifferent thermoreceptors in the body This was because

that during the previous research on the action mecha-nism of moxibustion we paid more attention to the effectsof moxibustion therapy on the diseased target organ andoverlooked the unique path and mechanism of startingeffects by moxibustion on the target organ Every effectivetherapeutic measure may produce effects on the target organand the specificity of its action path is the important basisfor existence of the method Based on the stimulation-response process of thermal stimulation by moxibustionrarr local reception rarr temperature rarr chemical conjuga-tion linkage rarr biological information cascade reaction rarreffect realization [19] we believed that due to differenttemperatures of thermal stimulation there are differences ofexcitation receptors the thermal stimulation at 46∘C mayactivate TRPV1 in the local site of moxibustion and promotedegranulation of Mast cells and the consequent effectsmay participate in regulation of cardiac function thereforethe thermal stimulation at 46∘C has better improvementeffects on cardiac function in brachycardia rats The thermalstimulation and the resulting inflammatory reaction are themost basic features ofmoxibustion effect [20] so in our studyattention was paid to the characteristics of the effects in thelocal site of moxibustion it may help in further researchof the paths of neurology endocrine and immune networkinteraction of moxibustion and also provide a new path toclarify the action mechanism of the effects of moxibustion


Abbreviations

AP Arterial pressure+dpdtmax Maximum rate of left ventricular pressure riseHR Heart rateLVSP Left ventricular systolic pressureMAP Mean arterial pressureMCs Mast cellsTRP Transient receptor potentialTRPV1 Transient receptor potential vanilloid

subfamily member 1t-dpdtmax Time from the beginning of left ventricular

systole to the maximum rate of left ventricularpressure rate

Acknowledgment

This present study was supported by the National Key BasicResearch Program (NKBRP) 2009CB522905 funded byMOST of China

References

[1] M J Gunthorpe C D Benham A Randall and J B DavisldquoThe diversity in the vanilloid (TRPV) receptor family of ionchannelsrdquo Trends in Pharmacological Sciences vol 23 no 4 pp183ndash191 2002

[2] M J Caterina A Leffler A B Malmberg et al ldquoImpairednociception and painsensation in mice lacking the capsaicinreceptorrdquo Science vol 288 no 5464 pp 306ndash313 2000

[3] J Huang Effect of TRPV1 on induceing release of inflammatorymediators in human keratinocytes and intervention drugs [PhDthesis] The Second Military Medical University 2009

[4] B I Toth TGeczy ZGriger et al ldquoTransient receptor potentialvanilloid-1 signaling as a regulator of human sebocyte biologyrdquoJournal of Investigative Dermatology vol 129 no 2 pp 329ndash3392009

[5] G-H He The synaptic effects of sympathetic histamine and itspathophysiological roles in acute ischemia induced arrhythmia[PhD thesis] The Fourth Military Medical University 2010

[6] X-Y Yu and Q Liu ldquoRoles of transient receptor potentialvanillod receptor subtype 1 in cardiovascular systemrdquo Advancesin Cardiovascular Disease vol 33 no 2 pp 282ndash285 2012

[7] R-L Guo N-X Zhao and Q-Y Ren ldquoInfluence of moxibus-tion therapy on fingerrsquos microcirculation and hemorrheology inhealth people after moxibustionrdquo Journal of the Fourth MilitaryMedical University vol 23 no 12 pp 1112ndash1114 2002

[8] N-X Zhao R-L Guo Q-Y Ren R-F Guo and N PanldquoInfluence of moxibustion of baihui (GV 20) on hemodynamicsof common carotid artery in healthy subjectsrdquo Zhen Ci Yan Jiuvol 32 no 4 pp 252ndash254 2007

[9] J-NHe andM F Luo ldquoProgress in the study on the relationshipbetween effects of acu-moxibustion andmast cells in acupointsrdquoZhen Ci Yan Jiu vol 32 no 3 pp 214ndash216 2007

[10] Y-J Shen S-P Ma W-L Deng et al Pharmacology of ChineseMateria Medica Peoplersquos Medical Publishing House 2000

[11] S-Y Xu X Chen and R-L BianMethodology of Pharmacolog-ical Experiment Peoplersquos Medical Publishing House 2006

[12] C-Y Chen B Xu X-C Yu et al ldquoStudy of the differencein the regulating effect on rat brachycardia between elec-troacupuncture at meridianpoints and corresponding non-meridian pointsrdquo Shanghai Journal of Acupuncture and Moxi-bustion vol 29 no 12 pp 747ndash751 2010

[13] B Xu X-C Yu C-Y Chen et al ldquoRelationship between efficacyof electroacupuncture and electroacupuncture stimulation ofdifferent acupoints and different tissue layers of acupoint areain hypotension plus bradycardia ratsrdquo Zhen Ci Yan Jiu vol 35no 6 pp 422ndash428 2010

[14] D Roosterman T Goerge S W Schneider N W Bunnett andM Steinhoff ldquoNeuronal control of skin function the skin as aneuroimmunoendocrine organrdquo Physiological Reviews vol 86no 4 pp 1309ndash1379 2006

[15] A J Stokes L M Shimoda M Koblan-Huberson C NAdra and H Turner ldquoA TRPV 2-PKA signaling module fortransduction of physical stimuli in mast cellsrdquo The Journal ofExperimental Medicine vol 200 no 2 pp 137ndash147 2004

[16] R M van den Wijngaard T K Klooker O Welting etal ldquoEssential role for TRPV1 in stress-induced (mast cell-dependent) colonic hypersensitivity in maternally separatedratsrdquo Neurogastroenterology amp Motility vol 21 no 10 p 1107-e94 2009

[17] B-Z ZhangHistophysiology ofMeridians andCollaterals Chan-nels Line China Association of Acupuncture and Moxibustion1987

[18] S J Galli ldquoNew concepts about the mast cellrdquoTheNew EnglandJournal of Medicine vol 328 no 4 pp 257ndash265 1993

[19] S-F LuH-Y Yin Y Tang et al ldquoConsiderations about study onmechanisms of thermal efficacies ofmoxibustion from activitiesof transient receptor potential familyrdquo Zhen Ci Yan Jiu vol 37no 2 pp 151ndash154 2012

[20] L-L Wang ldquoCharacteristic of moxibustion and its warming-dredging effectrdquo Zhongguo Zhen Jiu vol 31 no 10 pp 865ndash8682011

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Volume 2014Hindawi Publishing Corporationhttpwwwhindawicom


thermal stimulation at two temperatures below and above43∘C on cardiac function in rates as well as on skin in thelocal site of moxibustion whether there is any differenceor association of effects and whether they may provideany basis for optimizing the clinical operating program ofmoxibustion

2 Materials and Methods

21 Materials

211 Animals and Grouping Healthy male SD rats of cleangrade with body weight of 280ndash380 g Certificate no SCXK(Shanghai) 2007-0005 were purchased from Shanghai SlacLaboratory Animal Co Ltd and bred in batches Theexperiment was carried out in 1 week after the rats wereaccommodated The rats were divided into the followinggroups by the random number table Model Control GroupMoxibustionGroup 1 andMoxibustionGroup 2Thenumberof rats in each group would support to obtain 8 complete datasuccessfully During the experiment the animal treatmentmeasures were in strict accordance with the regulationsin the Instructive Opinions on Humanitarian Treatment ofLaboratory Animals issued in 2006 by theMinistry of Scienceand Technology of the Peoplersquos Republic of China and theprovisions for care and use of laboratory animals by NanjingUniversity of Traditional Chinese Medicine

212 Main Laboratory Equipment Moxa strips for animals(in the diameter of 7mm Nanyang Chinese Medicine MoxaCompany) thermostatic operating table for animals (ST-1model Chengdu Instrument Factory) multichannel physio-logical signal acquisition system (RM 6240 model ChengduInstrument Factory) temperature measuring device (HC-04 model Hangzhou Hongchang Technology Company)microsyringe pump (ALC-IP800 Shanghai Alcott BiotechCo Ltd) full automatic tissue processor (TP1020 GermanLEICA Company) paraffin slicing machine (RM2145 Ger-man LEICA Company) optical microscope (BX60 JapaneseOLYMPUS Company) and video camera (DP71 ImageAcquisition System Japanese OLYMPUS Company) wereused

22 Methods

221 Electrophysiological Experimental Method Fast a maleSD rat overnight for 12 hours with free access to water weighthe rat carry out anesthesia of the rat with 20 urethane(ethyl carbamate) in the dose of 1 gkg body weight fix the ratin the thermostatic operating table for animals and connectwith the electrocardiograph and the temperature transducer(via the anus of the rat) once the electrocardiogram andbody temperature of the rat are stable (37∘C) carry outintubation in the left ventricle as well as in the femoral arteryand femoral vein of the right lower limb to measure leftventricular pressure and arterial pressure and administer thedrug intravenously Model control group 10 minutes afterthe operation if the measurement indexes are stable start

recording data 5 minutes later rapid inject 04 inderal(4mgkg) via the femoral vein then slowly inject intra-venously to maintain inderal at the dose of 025mgkgminwith the microsyringe pump for 60min to establish thebradycardia model no any other interventions are carriedout Moxibustion group 1 establish the model as describedin the model control group At 20min during administrationof inderal at the maintenance dose ignite the customizedslender moxa strip (20 cm long and in the diameter of7mm) carry out moxibustion directly to stimulate at theXimen(PC4) Acupoint in the left upper limb of the rat for10min place the probe of the temperature controller atthe surface of the skin at the acupoint for moxibustion tomonitor real-time temperature and control the temperaturesin the local skin for moxibustion at 38∘C plusmn 1∘C (Figure 1(a))the flaming moxa strip and skin are 35 plusmn 5mm apartafter moxibustion continue observation for another 30minMoxibustion group 2 establish the model as described inthe model control group At 20min during administrationof inderal in the maintenance dose carry out moxibustionfor thermal stimulation of 10min at 46∘C carry out theintervention similar to that as described in moxibustiongroup 1 during thermal stimulation by moxibustion controlthe temperature in the local skin for moxibustion at 46∘C plusmn1∘C Figure 1(b) and the flaming moxa strip and skin are 12 plusmn2mm apart after moxibustion continue observation foranother 30min During moxibustion gently tap any moxaashes at the tip of the moxa strip to the ashtray based onthe combustion state of the moxa strip to always maintaincleanliness of the operating interface The Ximen Acupointin the left limb is selected the positioning method of theacupoint is the anthropomorphic comparative method thatis Ximen In the inner side at the 12 folding point of theforearm and in the middle of the gap between the radius andthe ulna The reason for the choice of the temperature is thatbased on our observations less than 38∘C thermal stimulusthe human body for the warm feeling is not obvious andabove 46∘C thermal stimulus the human body for the hotfeeling is very obvious difficult to continue to endure Duringthe whole course using RM6240B multichannel physiologyrecorder (parameters acquisition frequency 1 kz scan speed40msdiv and filter frequency 30Hz) to simultaneouslymonitor and record many indexes for cardiac function andhemodynamics including heart rate (min) mean femoralartery pressure (mmHg) left ventricular systolic pressure(mmHg) maximum rate of left ventricular pressure rise(mmHgs) and time (ms) from the beginning of left ventric-ular systole to the maximum rate of left ventricular pressurerate and so forth

222 Morphological Experimental Method

(1) Obtain Materials After completion of the monitoring andrecording of the electrophysiology experiment sacrifice therats and obtain materials Rapidly extract skin tissues in thelocal site at the XimenAcupoint in the rats of each groupwiththe ophthalmic forceps and the surgical scissors and placeinto 10 formaldehyde solution to store and fix for 1 week


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3 Results
















4 Discussion



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OncologyJournal of





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Abbreviations




Acknowledgment


References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















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Abbreviations




Acknowledgment


References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















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Abbreviations




Acknowledgment


References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of

















Abbreviations




Acknowledgment


References


























of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of





















of






Disease Markers



OncologyJournal of





PPAR Research



Journal of

ObesityJournal of
















Documents

Research on Effects of the Thermal Stimulation by Moxibustion at