ORIGINAL ARTICLE

Sinisan, a traditional Chinese medicine, attenuates experimentalchronic pancreatitis induced by trinitrobenzene sulfonic acidin rats

Yu Tang • Yonghui Liao • Nobuko Kawaguchi-Sakita •

Vikram Raut • Elham Fakhrejahani •

Niansong Qian • Masakazu Toi

Published online: 14 January 2011

� Japanese Society of Hepato-Biliary-Pancreatic Surgery and Springer 2010

Abstract

Background/purpose Sinisan, a traditional Chinese med-

icine, is effective for the treatment of gastrointestinal

disorders. In this study, we investigated the potential

protective role of Sinisan against chronic pancreatitis (CP)

in rats.

Methods CP was induced in rats by intrapancreatic

injection of trinitrobenzene sulfonic acid (TNBS). Rats

were randomly divided into a sham group, a TNBS-induced

CP group and a Sinisan-treated group. Serum amylase and

histological score were used to evaluate the severity of

disease. The levels of tumor necrosis factor-a (TNF-a),

interleukin-1b (IL-1b), cyclooxygenase-2 (COX-2), inter-

leukin-10 (IL-10) and a-smooth muscle actin (a-SMA) were

also measured in the three groups. Mechanical allodynia

was measured with von Frey filaments. In addition, the

protein levels of nerve growth factor (NGF) were measured

in pancreatic tissues.

Results Administration of Sinisan significantly decreased

the severity of CP. In the Sinisan-treated group, serum

amylase, TNF-a, IL-1b, COX-2 and a-SMA levels were

lower and the level of IL-10 was upregulated compared

with the TNBS-induced CP group. Furthermore, treatment

with Sinisan significantly, though not completely, attenu-

ated the allodynia. Simultaneously NGF expression was

also significantly downregulated in the Sinisan-treated

group compared with the TNBS-induced CP group.

Conclusions Sinisan could be an effective treatment

modality for CP via its anti-inflammatory, anti-fibrotic and

analgesic properties. It may be a promising drug candidate

for the treatment of patients with CP.

Keywords Traditional Chinese medicine � Sinisan �Chronic pancreatitis � Trinitrobenzene sulfonic acid

Introduction

Chronic pancreatitis (CP) is a pathophysiological event

of inflammation and necrosis resulting in fibrosis, pan-

creatic stone formation, diabetes mellitus and pain. The

mechanism by which this process occurs is not fully

understood and therapeutic strategies to treat CP are very

limited [1].

Traditional Chinese medicine (TCM) is a holistic

approach to health that brings the body, mind and spirit

into harmony. Increasing evidence indicates that several

herbs and their combinations used in TCM play important

pharmacological roles. The number of databases and

compilations of herbs, herbal formulations, phytochemical

Y. Tang and Y. Liao contributed equally to this work.

Y. Tang

Department of Medical Image Center,

Chinese PLA 302 Hospital, Beijing, China

Y. Liao

Department of Hepatobiliary Surgery, XiJing Hospital,

Fourth Military Medical University, Xi’an, China

N. Qian (&)

Department of Hepatobiliary Surgery,

Chinese People’s Liberation Army General Hospital,

Beijing 100853, China

e-mail: qianas@kuhp.kyoto-u.ac.jp

N. Kawaguchi-Sakita � V. Raut � E. Fakhrejahani � N. Qian �M. Toi (&)

Department of Breast Surgery, Graduate School of Medicine,

Kyoto University, Kyoto, Japan

e-mail: toi@kuhp.kyoto-u.ac.jp

123

J Hepatobiliary Pancreat Sci (2011) 18:551–558

DOI 10.1007/s00534-010-0368-z

constituents and molecular targets is growing, primarily

due to the widespread use of TCM. Herbal therapies are

mainly used for promoting overall good health, reputedly

with fewer side effects than Western-style remedies [2].

Some of the prescriptions have proven their efficacy at

pharmacological and biological levels. One example is

Sinisan, a traditional Chinese medical formula recorded by

Zhongjing Zhang in 200–201 AD. Sinisan, the major com-

ponents of which are saikosaponins, naringin, paeoniflorin

and glycyrrhizin [3], has been popular for treating various

diseases. Sun et al. [4] indicated that Sinisan may con-

tribute to the treatment of immunologically related diseases

by downregulating the activation and function of T lym-

phocytes, especially adhesion to type I collagen and matrix

metalloproteinase-2 and -9. In the contact dermatitis mouse

model, Sinisan significantly reduces tumor necrosis factor-

a (TNF-a), cyclooxygenase-2 (COX-2) and extracellular

signal-regulated kinase 1/2 phosphorylation [5]. It also

alleviates liver injury through protecting the hepatocyte

membrane, promoting nitric oxide release, inhibiting

lymphocyte activation and causing apoptosis of liver-

infiltrating cells [3]. In addition, Sinisan prevents the

progression of acute gastric mucosal lesions in rats by

attenuating enhanced neutrophil infiltration and lipid per-

oxidation associated with decreased vitamin E and ascorbic

acid and Se-glutathione peroxidase activity [6, 7]. More-

over, Sun et al. [8] demonstrated that trinitrobenzene sul-

fonic acid (TNBS)-induced colitis in mice was improved

noticeably by Sinisan, leading to a dramatic decrease in

pro-inflammatory cytokines including interferon-c (INF-c),

interleukin (IL)-12, TNF-a and IL-17 and an increase in

IL-10. Furthermore, a connection between acute pancrea-

titis or CP and ulcerative colitis has been reported [9, 10].

Pancreatitis is seen in 14–53% patients with ulcerative

colitis on autopsy. Although the pathogenesis of this

association is unclear, an autoimmune mechanism is one of

the causes [11]. These findings have prompted us to

explore the effectiveness of Sinisan in CP from an exper-

imental aspect. In this study, we examined the therapeutic

potential of Sinisan in the TNBS-induced CP model in rats,

which has been well-described in our previous study [12],

and its possible therapeutic mechanisms.

Materials and methods

All experimental procedures were approved by the Ani-

mal Use and Care Committee for Research and Educa-

tion of the PLA 302 Hospital (Beijing, People’s Republic

of China). The ethical guidelines were followed during

the experiment in conscious animals. All efforts were

made to minimize the number of animals used and their

suffering.

Experimental design

The animals were randomly allocated to three groups: a

TNBS-induced CP group (n = 30), a Sinisan-treated group

(n = 30) and a sham group (n = 20). After TNBS injec-

tion for 3 days, Sinisan (1000 mg/kg) p.o. for 25 days was

administered to the Sinisan-treated group. Simultaneously,

placebo was administered to the TNBS-induced CP group.

Behavioral tests were performed before induction of CP

and once weekly for up to 4 weeks afterwards. In the

strength–response study, rats were tested 4 weeks after CP

induction. At 4 weeks, surviving rats were killed by an

overdose of anesthesia, and blood samples were collected

for biochemical assays. Pancreases were rapidly removed

for histopathologic examination, Western blotting and

reverse transcriptase–polymerase chain reaction (RT-PCR)

(Fig. 1).

Drugs

The ethanol extract of Sinisan was made by the usual

method [5]. Briefly, equal amounts of each material

(saikosaponins, naringin, paeoniflorin and glycyrrhizin)

were mixed and extracted twice with 70% ethanol under

reflux for 1 h. After filtration and reclaiming the ethanol,

the extract was lyophilized to obtain a powder. The powder

was dissolved in normal saline (100 mg/ml) for in vivo

assay by gavage oral administration to rats. The same

volume of normal saline was used as placebo.

Induction of CP

Our TNBS-induced rat CP model is described elsewhere

[12]. Male Sprague–Dawley rats (250–300 g) were used

in this study. Animals were given free access to drinking

water and standard food pellets. Food was withdrawn

12 h prior to induction of CP. Briefly, the common bile

duct was closed temporarily near the liver with a small

vascular clamp. A blunt 28 gauge needle with PE 10

tubing attached was inserted into the duodenum and was

guided through the papilla into the duct and was secured

with a suture. TNBS solution (0.5 ml, 2%) in 10% etha-

nol in phosphate-buffered saline (PBS, pH 7.4) was

infused into the pancreatic duct over a period of 2–5 min

at a pressure of 50 mmHg. After 30 min exposure to

Fig. 1 Schematic description of the methods used in the article

552 J Hepatobiliary Pancreat Sci (2011) 18:551–558

123

TNBS, needle and tubing were removed. The hole in the

duodenum was sutured and the vascular clamp was

removed, restoring the bile flow. All the procedures in the

sham group were same as those in the TNBS group,

except that the same volume of saline instead of TNBS

was infused into the duct.

Behavioral tests

The development of referred pain during the course of CP

was assessed by the abdomen response frequencies (RFs)

between 1 and 4 weeks after induction of CP. Mechanical

allodynia was measured with von Frey filaments (VFF)

(Stoelting, Kiel, USA). Testing was performed according

to our previously reported protocol [12]. Briefly, the belly

was shaved prior to the test and areas designated for

stimulation were marked. Rats were placed in a plastic

cage with a mesh floor and were given 30 min for adap-

tation before testing. The VFF were applied from below

through the mesh floor, in ascending order to the

abdominal area at different points on the surface. The

withdrawal response to mechanical stimuli applied to the

abdomen was assessed by 6 calibrated VFF (2.29, 2.75,

6.76, 16.6, 40.7 and 69.2 mN). VFF of various calibers

were applied in an ascending order to the abdomen 10

times each for 1–2 s with a 10 s interval between the

applications. Mechanical hypersensitivity in the abdominal

area was quantified by measuring the number of with-

drawal responses. The data are expressed as a percentage

of the number of positive responses to each filament for

each rat. All the tests were performed in a blinded fashion.

In the time-course study, behavioral tests were performed

before and once weekly for up to 4 weeks after induction

of CP with a single force of VFF (40.7 mN). In the

strength–response study, rats were tested 4 weeks after

pancreatitis induction with a series of VFF (2.29, 2.75,

6.76, 16.6, 40.7 and 69.2 mN).

Pancreatic histology

The histology of the rat pancreas was visualized with

hematoxylin and eosin stain (H&E). Rats were deeply

anesthetized with sodium pentobarbital (60 mg/kg i.p.),

and the pancreatic tissues were obtained and then fixed in

4% paraformaldehyde in phosphate buffer (PB, pH 7.4) at

4�C overnight. Pancreatic tissue was then transferred to

xylene washes, placed in cassettes and embedded in par-

affin. Paraffin blocks were cut into 5-lm sections and

stained with H&E. A pathologist analyzed the histological

sections in a double-blinded manner. The severity of CP

was morphologically assessed by semiquantitative graded

scores: oedema (0–3), necrosis (0–3), inflammatory cell

infiltration (0–3) and fibrosis (0–3), as reported previously

[13].

RT-PCR analysis

RT-PCR was performed to analyze the mRNA expression

levels of TNF-a, IL-1b, COX-2 and IL-10 in the pancreatic

tissue. RNA was extracted from pancreatic tissue using

Trizol reagent (Invitrogen, Carlsbad, USA). One micro-

gram of RNA was reverse transcribed to cDNA. The

primer sequences used in PCR were as follows: GAPDH,

50-AACGACCCCTTCATTGAC and 30-CACGACTCATA

CAGCACCT; TNF-a, 50-CATCTTCTCAAAATTCGAGT

GACAA and 30-CCCAACATGGAACAGATGAGGGT;

IL-1b, 50-GCCCATCCTCTGTGACTC and 30-CGTGGAG

TGTTCGTCTCG; COX-2, 50-ATCACTCAGTTTGTTGA

GTCATTC and 30-GTAATTGGGATGTCATGATTAG;

IL-10, 50-GGCTCAGCACTG CTATGTTGCC and 30-AG

CATGTGGGTCTGGCTGACTG. Amplification was per-

formed with the following cycles: 95�C for 30 s, followed

by 40 cycles of denaturing at 95�C for 5 s and annealing at

60�C for 20 s. All of the reactions were performed in

triplicate.

Western blotting

Protein extracts from pancreatic tissue were resolved by

sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE)

and transferred to a nitrocellulose membrane (Amersham,

Piscataway, USA). The membrane was blocked and probed

with monoclonal antibody to a-smooth muscle actin

(a-SMA; 1:300; Sigma, St. Louis, USA), then incubated

with horseradish peroxidase-labeled second antibody.

Protein band intensities were quantified using Image Pro

Plus 5.0. All of the reactions were performed in triplicate.

Biochemical assays

Serum TNF-a, IL-1b, IL-10 and nerve growth factor (NGF)

were determined by ELISA. The serum TNF-a, IL-1b and

IL-10 levels were evaluated using ELISA kits (Boster,

Wuhan, China). NGF protein content in pancreatic tissues

was measured using an NGF Emax immunoassay system

ELISA kit (Promega, Madison, USA). The serum amylase

level was determined by a Beckman CX7 Chemistry

Analyzer (Beckman, Fullerton, USA).

Statistical analysis

All results shown represent mean ± SD. Statistical analy-

ses were performed using an unpaired, two-tailed Student’s

t-test with a significance level of P \ 0.05.

J Hepatobiliary Pancreat Sci (2011) 18:551–558 553

123

Results

Effects of Sinisan on microscopic changes

and histological score in the TNBS-induced

CP group

The survival rate of the TNBS-induced CP group at

4 weeks was 93.3% (1 died in week 1 and 1 died in

week 3). In contrast, all the rats in the Sinisan-treated

group and the sham group survived for the duration of the

study period. In the sham group, the pancreas was his-

tologically normal. However, 4 weeks after TNBS injec-

tion, the pancreas showed significant oedema, necrosis,

fibrosis, stromal proliferation and inflammatory infiltra-

tion. Compared with these changes, Sinisan significantly

improved all the above parameters. Figure 2a–c shows

representative photos of H&E staining for pancreas

tissues in the sham group, the TNBS-induced CP group

and the Sinisan-treated group. The changes in score for

oedema, necrosis, inflammatory cell infiltration and

fibrosis are shown in Fig. 2d.

Effects of Sinisan on TNF-a, IL-1b, COX-2, IL-10

and a-SMA in pancreatic tissues after

TNBS-induced CP

Pancreatic tissue mRNA expression of TNF-a, IL-1b and

COX-2 were significantly increased at 4 weeks after the

onset of CP relative to the sham group. In addition, treat-

ment with Sinisan markedly decreased the TNBS-induced

increases in TNF-a, IL-1b and COX-2 mRNA expression

(Fig. 3). Also, treatment with Sinisan significantly down-

regulated the protein levels of a-SMA in pancreatic tissue

which was unregulated by TNBS injection (Fig. 4). Further

analysis of anti-inflammatory cytokines showed a signifi-

cant upregulation of IL-10 mRNA expression in the TNBS-

Fig. 2 Effects of Sinisan on the

microscopic changes and

histological scores 4 weeks

after TNBS injection. H&E

staining of the pancreas in

(a) the sham group (n = 20);

(b) the TNBS-induced CP group

(n = 28); and (c) the Sinisan-

treated group (n = 30). d The

severity of CP in the above

groups was morphological

assessed by semiquantitative

scoring. *P \ 0.05 compared

with the sham group; #P \ 0.05

compared with the TNBS-

induced CP group

554 J Hepatobiliary Pancreat Sci (2011) 18:551–558

123

induced CP group compared to the sham group; treatment

with Sinisan led to a further increase in mRNA expression

of IL-10 (Fig. 3).

Effects of Sinisan on serum amylase, TNF-a, IL-1band IL-10 after TNBS-induced CP

Intrapancreatic injection of TNBS resulted in significant

increases in serum amylase at 4 weeks (P \ 0.01). Com-

pared with the TNBS-induced CP group, the group treated

with Sinisan had significantly reduced serum amylase

levels, down to those of the sham group (P \ 0.01). Fur-

thermore, serum IL-1b and TNF-a levels in the Sinisan-

treated group were significantly decreased in comparison

with those of the TNBS-induced CP group (P \ 0.01 and

\0.05, respectively). In addition, the serum IL-10 level

was increased significantly in the Sinisan-treated group

(P \ 0.01, vs. TNBS-induced CP group; Table 1).

Effects of Sinisan on mechanical allodynia

in the TNBS-induced CP group

It was found in the present study that allodynia in the

TNBS-induced CP group was significantly increased com-

pared to that of the sham group at the same designated time

points, from 1 to 4 weeks. RFs were significantly higher in

the TNBS-induced CP group than in the sham group after

1 week (RFs = 38.2 ± 3.9%, P \ 0.05) and persisted for

4 weeks (RFs = 67.6 ± 4.6%, P \ 0.05) as compared with

sham group (Fig. 5a). In addition, the RFs to VFF stimu-

lation curve in the TNBS-induced CP group was statisti-

cally significantly left of the RFs to VFF stimulation curve

in the sham group (Fig. 5b). Thus, we confirmed a signifi-

cant increase in sensitivity to mechanical probes in the

abdomen. NGF, which is linked to the development of

allodynia in CP, was significantly upregulated compared

with the sham group at 4 weeks (P \ 0.01, Fig. 5c).

In order to ascertain the effects of Sinisan on mechanical

Fig. 3 Effects of Sinisan on the mRNA expression changes of

TNF-a, IL-1b, COX-2 and IL-10. a Changes of TNF-a, IL-1b, COX-2

and IL-10 mRNA levels were determined in the sham group (n = 20),

the TNBS-induced CP group (n = 28) and the Sinisan-treated group

(n = 30). b The relative mRNA levels are shown in the diagram.

*P \ 0.05 compared with the sham group; #P \ 0.05 compared with

the TNBS-induced CP group

Fig. 4 Effects of Sinisan on the protein expression changes of

a-SMA. a Changes in a-SMA protein levels were determined in the

sham group (n = 20), the TNBS-induced CP group (n = 28) and the

Sinisan-treated group (n = 30). b The relative mRNA levels are

shown in the diagram. *P \ 0.05 compared with the sham group;#P \ 0.05 compared with the TNBS-induced CP group

J Hepatobiliary Pancreat Sci (2011) 18:551–558 555

123

allodynia in rats with TNBS-induced CP, we also tested the

RFs to VFF on the Sinisan-treated group at 4 weeks. The

results showed that treatment with Sinisan significantly,

though not completely, attenuated the allodynia (P \ 0.05,

Fig. 5b). NGF protein levels were also significantly

decreased compared with the TNBS-induced CP group

(P \ 0.05, Fig. 5c).

Discussion

Sinisan is effective for improving disorders of the digestive

system including liver injury [3], contact dermatitis [5] and

colitis [8]. In China, Sinisan is already used to treat patients

with CP by some traditional Chinese doctors and has shown

some curative effects [14–16]. However, the mechanism is

still unclear. In this study, therefore, we investigated the

mechanism of Sinisan in the treatment of a TNBS-induced

CP model. Serum amylase and histological score reflect the

extent of pancreatic tissue injury. Our results showed that

the above-mentioned indexes were significantly improved

in the Sinisan-treated group. In addition, the pro-inflam-

matory and anti-inflammatory cytokine response in CP was

significantly altered [17]. As representatives of pro-

inflammatory cytokines, TNF-a, IL-1b and COX-2 have

been shown to play a major role in the pathophysiology of

the inflammatory diseases including CP. TNF-a is produced

by activated macrophages, endothelial cells and B lym-

phocytes with multiple functions and plays important roles

in CP [18]. Marrache et al. [19] demonstrated that the

overexpression of IL-1b in the murine pancreas could

induce CP, highlighting the potential importance of IL-1bin immune activation in the pancreas. Overexpression of

COX-2 in CP modulates the degree of pancreatic inflam-

mation and affects both the exocrine and the endocrine

pancreas [20]. Our present study revealed that TNBS-

induced CP is accompanied by a significant increase in the

pancreatic mRNA expression of TNF-a, IL-1b and COX-2

and an increased release of these cytokines in serum. In

addition, treatment with Sinisan significantly suppressed the

gene expression of TNF-a, IL-1b and COX-2 in pancreatic

tissue and, decreased serum levels of TNF-a, IL-1b and

COX-2 during the course of CP. These results indicate that

Sinisan effectively alleviates the injury caused by activation

of the immune system during CP. In contrast to these pro-

inflammatory cytokines, IL-10 is a potent anti-inflammatory

cytokine, which is known to limit the severity of inflam-

mation by downregulating pro-inflammatory mediator

release and has immunosuppressive effects [21]. IL-10 also

has direct anti-proliferative and anti-fibrotic properties [22].

In our study, we observed a moderate increase in the serum

IL-10 level and a significant increase in IL-10 mRNA

expression after the development of CP. The administration

of Sinisan not only significantly increased the serum IL-10

level, but also further increased the mRNA expression of

IL-10. In addition, the fibrosis score and tissue a-SMA level

at 4 weeks in the Sinisan-treated group were significantly

lower than in the TNBS-induced CP group, suggesting that

Sinisan also prevents pancreatic fibrosis in addition to

inflammation. Furthermore, treatment with Sinisan signi-

ficantly, though not completely, attenuated the allodynia.

NGF belongs to the neurotrophin family. In somatic pain

models, increase in NGF is linked to the development of

allodynia. NGF production in the acute pancreatitis is

responsible for plastic changes in the sensory neurons

that mediate peripheral sensitization and contribute to the

generation of pain [23]. Friess et al. [24] indicated that the

NGF/TrkA pathway might also be directly involved in pain

generation in CP. In our study, we observed a significant

increase in NGF protein expression after the development

of CP. The administration of Sinisan significantly decreased

the NGF level, indicating that a possible mechanism of the

Sinisan effect on CP-induced pain is that it inhibits NGF

expression.

As mentioned above, saikosaponins, naringin, paeoni-

florin and glycyrrhizin are the major components of Sinisan.

Saikosaponins are also the main component of Saiko-

keishi-to which has been widely used in Japan as an anti-

inflammatory agent for patients with CP [25]. Su et al.

[26, 27] reported that Saiko-keishi-to has anti-fibrotic and

anti-inflammatory effects on CP by downregulating trans-

forming growth factor b1 and pancreatitis-associated pro-

tein. In addition, severe acute pancreatitis in rats could

obviously impact the absorption and pharmacokinetic

parameters of naringin, indicating its potential therapeutic

role [28]. Moreover, paeoniflorin could inhibit the function

Table 1 Comparison of serum amylase, TNF-a, IL-1b and IL-10 in each group

Sham group (n = 20) TNBS-induced CP group (n = 28) Sinisan-treated group (n = 30)

Serum amylase (U/l) 993 ± 64.7 1763 ± 100.6a 1134 ± 424.1c

TNF-a (pg/ml) 52.5 ± 13.1 193.2 ± 58.8a 122.6 ± 55.9a,c

IL-1b (pg/ml) 89.5 ± 25.4 193.2 ± 39.9a 140.2 ± 58.1b,d

IL-10 (pg/ml) 69.7 ± 16.9 72.7 ± 3.8 268.9 ± 22.4a,c

a P \ 0.01, b P \ 0.05, versus control group; c P \ 0.01, d P \ 0.05, versus TNBS-induced CP group

556 J Hepatobiliary Pancreat Sci (2011) 18:551–558

123

of IL-1b by suppressing b-actin and Hsp27 phosphorylation

in rat brain microvessel endothelial cells [29]. Paeoniflorin

also exerted negative regulation on macrophage function

including downregulating COX-2 and TNF-a [5]. In our

opinion, the main element which has a specific effect is

glycyrrhizin. Some publications have shown its anti-

inflammatory effect via suppression of TNF-a, NK-jB,

signal transducer and activator of transcription-3, and inter-

cellular adhesion molecule-1, and inactivation of PI3K/Akt/

GSK3b signaling and glucocorticoid receptor, etc. [30–33].

As we know, TNF-a is a key factor leading to the triggering

of an inflammatory cascade involving the induction of

cytokines such as INF-c and IL-1b [34]. NGF is produced in

a number of different cell types in response to TNF-a and

IL-1b; blocking NGF suppresses the marked thermal and

mechanical hyperalgesia induced by TNF-a or IL-1b [35].

So we have speculated that glycyrrhizin could downregu-

late NGF expression in CP by blocking TNF-a and IL-1b. It

is not clear whether there is an independent mechanism and

this will require further investigations. In addition, two

reports in the literature have revealed that treatment with

glycyrrhizin increases the production of IL-10 and sup-

presses T-cell mediated inflammation [36, 37]. Moreover,

Wu et al. [38] showed that glycyrrhizin significantly

decreased the expression of a-SMA in the TNBS-induced

CP model. Interestingly, in the TNBS-induced colitis

model, the therapeutic effect of Sinisan was much greater

than that of its content of glycyrrhizin, indicating that the

effect of Sinisan are produced by a combination of the

various active components contained in the formula [8].

Detailed studies should be performed to determine the

interactions between the various active ingredients.

In summary, the present study provided evidence for the

alleviating effects of Sinisan against CP through multiple

mechanisms, including reducing inflammation, preventing

pancreatic fibrosis and blocking the development of allo-

dynia. Therefore, we conclude that treatment with Sinisan

has therapeutic potential in the TNBS-induced CP model

and that it may be a promising drug candidate for the

treatment of patients with CP.

Acknowledgments This work was supported by Chinese Govern-

ment Scholarship no. 2009659015.

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