OBJECTIVETo explore the role and mechanism of oxidative inflammatory cascade in pancreatic fibrosis progression of chronic pancreatitis (CP) in mice induced by dibutyltin dichloride (DBTC) plus ethanol.

METHODSThirty-six KM mice were randomly divided into 2 groups (n = 18): control group and model group (DBTC combined with ethanol). The mice in model group were intravenously injected with DBTC (8 mg/kg) in tail vein and drink 10% ethanol. After modeling 2 weeks, 4 weeks and 8 weeks, the mice were anesthetized and sacrificed, the pathological changes and the degree of fibrosis in the pancreas were observed by HE and Masson staining, the F4/80 expression level were detected by immunohistochemistry, the content of superoxide dismutase (SOD), malondialdehyde(MDA) and myeloperoxidase (MPO) were measured in the pancreatic homogenates.

RESULTSThe fibroblasts and macrophages (f4/80 positive staining) could be seen obviously in pancreas of model group at 2 weeks. At 4 weeks and 8 weeks, macrophages infiltration increased and pancreatic tissue was substituted by the proliferation of fibrosis significantly. At every time-point, in pancreatic homogenates SOD was decreased, MDA and MPO markedly increased. There was significant differences between two groups (P < 0.05).

CONCLUSIONDBTC injection joint ethanol drinking can successfully establish the model of chronic pancreatitis and pancreatic fibrosis in mice. Oxidative inflammatory cascade plays an important role in the progression of pancreatic fibrosis.

Animals ; Disease Progression ; Ethanol ; adverse effects ; Fibrosis ; Immunohistochemistry ; Malondialdehyde ; metabolism ; Mice ; Organotin Compounds ; adverse effects ; Oxidative Stress ; Pancreas ; pathology ; Pancreatitis, Chronic ; chemically induced ; physiopathology ; Peroxidase ; metabolism ; Superoxide Dismutase ; metabolism

BACKGROUND/AIMS: To investigate the expression of Toll-like receptor 4 (TLR4) in the pancreases of rats with acute necrotizing pancreatitis (ANP) and any changes upon treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor kappaB (NF-kappaB), as well as to determine the relationship between TLR4 and NF-kappaB in ANP pathogenesis. METHODS: A total of 72 SD rats were randomly divided into three groups, namely, the control (sham-operation), ANP, and ANP with PDTC pretreatment groups. The PDTC-pretreated group was intraperitoneally injected with PDTC at a dose of 100 mg/kg 1 hour before the induction of ANP. The expressions of TLR4 and NF-kappaB in pancreatic tissue were evaluated by immunohistochemistry and Western blot analysis. The mRNA levels of cytokines tumor necrosis factor alpha, interleukin (IL)-1beta, and IL-6 were measured by reverse transcription polymerase chain reaction. RESULTS: The expressions of TLR4, NF-kappaB, and cytokine (NF-kappaB target) genes in the pancreatic tissue increased more significantly in the ANP groups than in the sham-operation group at 3, 6, and 12 hours. Pretreatment with PDTC alleviated the inflammatory activation in the pancreas with ANP, causing a significant decrease in the expressions of TLR4, NF-kappaB, and cytokine genes in the pancreatic tissue. CONCLUSIONS: The expressions of TLR4 and NF-kappaB were increased in the pancreases of rats with ANP. PDTC not only inhibits NF-kappaB but also suppresses the expression of TLR4 and downregulates the expression of the related cytokine genes.
Animals ; Antioxidants/*pharmacology ; Interleukin-1beta/genetics/metabolism ; Interleukin-6/genetics/metabolism ; Male ; NF-kappa B/*drug effects/metabolism ; Pancreas/metabolism/pathology ; Pancreatitis, Acute Necrotizing/chemically induced/*drug therapy ; Pyrrolidines/*pharmacology ; RNA, Messenger/metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Thiocarbamates/*pharmacology ; Toll-Like Receptor 4/*drug effects/metabolism ; Tumor Necrosis Factor-alpha/genetics/metabolism

BACKGROUND/AIMS: To investigate the expression of Toll-like receptor 4 (TLR4) in the pancreases of rats with acute necrotizing pancreatitis (ANP) and any changes upon treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of nuclear factor kappaB (NF-kappaB), as well as to determine the relationship between TLR4 and NF-kappaB in ANP pathogenesis. METHODS: A total of 72 SD rats were randomly divided into three groups, namely, the control (sham-operation), ANP, and ANP with PDTC pretreatment groups. The PDTC-pretreated group was intraperitoneally injected with PDTC at a dose of 100 mg/kg 1 hour before the induction of ANP. The expressions of TLR4 and NF-kappaB in pancreatic tissue were evaluated by immunohistochemistry and Western blot analysis. The mRNA levels of cytokines tumor necrosis factor alpha, interleukin (IL)-1beta, and IL-6 were measured by reverse transcription polymerase chain reaction. RESULTS: The expressions of TLR4, NF-kappaB, and cytokine (NF-kappaB target) genes in the pancreatic tissue increased more significantly in the ANP groups than in the sham-operation group at 3, 6, and 12 hours. Pretreatment with PDTC alleviated the inflammatory activation in the pancreas with ANP, causing a significant decrease in the expressions of TLR4, NF-kappaB, and cytokine genes in the pancreatic tissue. CONCLUSIONS: The expressions of TLR4 and NF-kappaB were increased in the pancreases of rats with ANP. PDTC not only inhibits NF-kappaB but also suppresses the expression of TLR4 and downregulates the expression of the related cytokine genes.
Animals ; Antioxidants/*pharmacology ; Interleukin-1beta/genetics/metabolism ; Interleukin-6/genetics/metabolism ; Male ; NF-kappa B/*drug effects/metabolism ; Pancreas/metabolism/pathology ; Pancreatitis, Acute Necrotizing/chemically induced/*drug therapy ; Pyrrolidines/*pharmacology ; RNA, Messenger/metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Thiocarbamates/*pharmacology ; Toll-Like Receptor 4/*drug effects/metabolism ; Tumor Necrosis Factor-alpha/genetics/metabolism

OBJECTIVETo explore the protective effects and mechanisms of electroacupuncture (EA) at Zusanli (ST36) acupoint in rats with severe acute pancreatitis (SAP).

METHODSSixty-six male Sprague-Dawley rats were randomly assigned to three groups of 22 each: a SAP model group (SAP group), a shamoperated group (sham group) and a EA at ST36 acupoint group (EA group). A rat model of SAP was induced by pancreatic duct injection with 3.5% sodium taurocholate. EA was performed at ST36 acupoint for 30 min after induction of SAP and 30 min before sacrificed. The rats were killed at 3 h (n=7), 6 h (n=7) and 12 h (n=8) after operation, and blood samples were taken for the measurement of tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6) and acetylcholine (Ach) by enzyme-linked immunosorbnent assay. The pathological changes of pancreatic tissue, volume of ascites and pancreatic weight/body weight ratio were measured.

RESULTSThe serum concentrations of TNF-α and IL-6 in the EA group were significantly lower than in the SAP group at 3, 6 and 12 h after the operation (p<0.05). Serum Ach in the EA group was significantly higher than in the SAP group at various time points after operation (p<0.05). The other parameters were clearly improved after treatment with EA.

CONCLUSIONEA at ST36 acupoint might have a therapeutic effect in rats with SAP through activating the cholinergic anti-inflammatory pathway.

Acetylcholine ; metabolism ; Acute Disease ; Animals ; Electroacupuncture ; Enzyme-Linked Immunosorbent Assay ; Interleukin-6 ; metabolism ; Male ; Pancreatitis ; chemically induced ; therapy ; Rats ; Rats, Sprague-Dawley ; Severity of Illness Index ; Taurocholic Acid ; toxicity ; Tumor Necrosis Factor-alpha ; metabolism

OBJECTIVETo observe the effects of Qingyi Granule (QYG) on the changes of total protein expressions in the pancreatic tissue of rats with severe acute pancreatitis (SAP) induced by sodium taurocholate (STC).

METHODSSAP was induced by retrograded injecting 5% STC from the gut-pancreatic duct in 36 Sprague-Dawley (SD)rats. Then they were randomly divided into the SAP group and the QYG treatment group (abbreviated as the QYG group), 18 in each group. After successful modeling, rats in the QYG group were administered with QYG water solution (W: W = 1:1) once with an interval of 12 h (1 mL/100 g), while rats in the SAP group were administered with normal saline. The medication was performed four times. The total proteins were extracted from the pancreatic tissue of all rats to perform two-dimensional electrophoresis, fluorescent staining, and atlas analysis. The protein dots with differential expressions more than four times between each other in 48 h gel pictures were chosen and used for MALDI-TOF/TOF mass chromatographic analysis and biological information analysis.

RESULTSThe 5% STC induced SAP model rats had typical pathological changes in the pancreatic tissue. The proteomics changes of the pancreatic tissue were analyzed by gel image manipulation software. Twenty two disparate points were detected between two groups at 48 h, 5 points of the protein were up-regulated and 17 points were down-regulated of the total after QYG intervention. Nine protein spots expressed differently more than 4 times and stably at 48 h, 7 kinds of proteins have been identified by mass chromatographic analysis and Data Base Retrieval, and they were Serpinb1a 39 kDa protein, Serpinb1a 43 kDa protein, Prdx4 Prx IV, Clps, gamma-actin (Actg1), Eprs and Hadhsc. Those proteins were involved in signal transmit during the process of SAP pancreas--pathological injury analyzed from their functions.

CONCLUSIONSProteomics can well reflect the effects of QYG on differential expression proteins in the pancreatic tissue of rats with SAP. Studying differential expression proteins may provide a new theoretical basis and molecule target for QYG treating SAP.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Pancreas ; metabolism ; Pancreatitis ; chemically induced ; metabolism ; prevention & control ; Proteome ; Rats ; Rats, Sprague-Dawley

In Korea, the use of herbal remedies is a common cause of drug-induced liver injury. However, the occurrence of both acute pancreatitis and acute hepatitis after taking herbal remedies has rarely been reported. Herein, we report a case of concurrent acute pancreatitis and acute hepatitis associated with Ceramium kondoi ingestion. A 58-year-old woman was diagnosed with advanced gastric cancer 7 months ago. Total gastrectomy and adjuvant chemotherapy was performed without complications. The patient had been well until recently, when she presented with severe abdominal pain after ingestion of Ceramium kondoi for 4 weeks. The laboratory findings demonstrated elevated liver enzymes and lipase, and abdominal computed tomography revealed pancreas swelling with fat infiltration. The diagnosis was made based on the diagnostic criteria for drug induced pancreatitis and the Russel Uclaf Causality Assessment Method scale for drug-induced liver injury. After cessation of Ceramium kondoi, she showed clinical and biochemical improvement.
Abdominal Pain/etiology ; Acute Disease ; Drug-Induced Liver Injury/*diagnosis/enzymology ; Female ; Humans ; Lipase/metabolism ; Liver/*drug effects ; Middle Aged ; Pancreas/*drug effects ; Pancreatitis/*chemically induced/*diagnosis ; Plant Extracts/chemistry/*toxicity ; Rhodophyta/chemistry/metabolism ; Tomography, X-Ray Computed

OBJECTIVETo investigate the effect of resveratrol on the apoptosis of pancreatic acinar cells in rats with severe acute pancreatitis (SAP) and explore the mechanism of such effect.

METHODSD rats with 3.5% sodium taurocholate-induced SAP were treated with resveratrol, and the serum amylase was detected with automatic biochemistry analyzer. The apoptosis of the pancreatic acinar cells in the rats was detected by TUNEL assay, and the expression of Fas and FasL genes was determined by RT-PCR and Western blotting. The pathological changes of the pancreas were observed under optical microscope.

RESULTSCompared with SAP group, the resveratrol-treated rats showed obviously decreased serum amylase and scores for pancreatic histopathological lesions. Resveratrol treatment significantly increased the apoptotic indices of pancreatic acinar cells and the levels of FasL mRNA and protein in rats with SAP.

CONCLUSIONResveratrol produces important therapeutic effect on SAP in rats by inducing pancreatic acinar cell apoptosis possibly as a result of up-regulated FasL gene expression.

Animals ; Apoptosis ; drug effects ; Fas Ligand Protein ; drug effects ; genetics ; metabolism ; Male ; Pancreas, Exocrine ; pathology ; Pancreatitis, Acute Necrotizing ; chemically induced ; drug therapy ; pathology ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; therapeutic use ; Taurocholic Acid ; Up-Regulation

OBJECTIVETo investigate the protective effect of resvertrol on the intestinal mucosal cells in rats with severe acute pancreatitis and explore the possible mechanism.

METHODSTwenty-four SD rats were randomly divided into the sham-operation (SO) group, severe acute pancreatitis (SAP) group and resveratrol-treated (RES) group. In the SO group, the pancreases were slightly flipped only. In the SAP and RES groups, SAP model was established by retrograde injection of 40 g/L sodium chrolate (1 ml/kg) through the pancreatic duct, and in the latter group, resveratrol (10 mg/kg) was given intravenously. Specimens were obtained 6 h after SAP model establishment and the endotoxin levels in the portal vein was determined with turbidimetry to evaluate the effect of resversatrol on the intestinal endotoxin translocation in SAP rats. Apoptosis of the mucosal cells was detected by TUNEL methods, and the expression of bax and bcl-2 mRNA were determined by RT-PCR. The mitochondrial membrane potential of the intestinal mucosal cells was measured by confocal microscopy.

RESULTSThe endotoxin levels in the portal vein were significantly lower in RES group than in SAP group (P<0.01). TUNEL assay demonstrated significantly higher apoptotic index of the mucosal cells in SAP group than that in RES group (P<0.01). The expression of Bax mRNA in the intestinal mucosal cell was significantly higher in SAP group than in RES group (P<0.01), whereas the expression of bcl-2 mRNA was significantly lower in SAP group (P<0.01). The mitochondrial membrane potential of the intestinal mucosal cell was significantly lower in SAP group than in RES group (P<0.01).

CONCLUSIONResvertrol can inhibit the apoptosis of the intestinal mucosa cells and maintain the integrity of the intestinal barrier to prevent the bacterial and endotoxin translocation in SAP.

Animals ; Anti-Inflammatory Agents, Non-Steroidal ; pharmacology ; therapeutic use ; Apoptosis ; drug effects ; In Situ Nick-End Labeling ; Intestinal Mucosa ; drug effects ; metabolism ; pathology ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Microscopy, Confocal ; Pancreatitis, Acute Necrotizing ; chemically induced ; drug therapy ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Sodium Chloride ; Stilbenes ; pharmacology ; therapeutic use ; bcl-2-Associated X Protein ; genetics

A stable and reliable infected necrotizing pancreatitis (INP) model in rats was established in order to study the pathophysiological mechanism and pathological development rule of INP and explore the new therapeutic methods for the diseases. Forty-six SD rats were randomly divided into 5 groups. The animals in group A received the injection of 5% sodium taurocholate into the pancreatic duct and those in group B underwent that of E. coli into the pancreatic duct. The rats in groups C, D and E were subjected to the injection of 5% sodium taurocholate in combination with different concentrations of E. coli (10(3), 10(4), 10(5)/mL, respectively) into the pancreatic duct. The dose of injection was 0.1 mL/100 g and the velocity of injection was 0.2 mL/min in all the 5 groups. Eight h after the injection, the survival rate of animals was recorded and the surviving rats were killed to determine the serum content of amylase and perform pathological examination and germ cultivation of the pancreatic tissue. The results showed that acute necrotizing pancreatitis model was induced by injection of 5% sodium taurocholate into the pancreatic duct. The positive rate of germ cultivation in group A was 12.5%. The acute necrotizing pancreatitis model was not induced by injection of E. coli into the pancreatic duct and the positive rate of germ cultivation in group B was 0. The INP model was established in groups C to E. The positive rate of germ cultivation was 60%, 100% and 100% and 8-h survival rate 100%, 100% and 70% in groups C, D and E, respectively. It was concluded that a stable and reliable model of INP was established by injection of 5% sodium taurocholate in combination with 10(4)/mL E. coli into the pancreatic duct with a dose of 0.1 mL/100 g and a velocity of 0.2 mL/min. The pathogenesis of INP might be that the hemorrhage and necrosis of pancreatic tissue induced by sodium taurocholate results in weakness of pancreatic tissue in fighting against the germs. Meanwhile, the necrotic pancreatic tissue provides a good proliferative environment for the germs.
Cholagogues and Choleretics/*pharmacology ; Disease Models, Animal ; Escherichia coli/*metabolism ; Injections, Intraperitoneal ; Pancreas/enzymology ; Pancreas/microbiology ; Pancreatic Ducts/enzymology ; Pancreatic Ducts/microbiology ; Pancreatitis, Acute Necrotizing/*chemically induced ; Pancreatitis, Acute Necrotizing/*microbiology ; Rats, Sprague-Dawley ; Taurocholic Acid/*pharmacology ; Time Factors

OBJECTIVETo study the expressions of Fas and Fas-Ligand (FasL) genes in rats with acute pancreatitis (AP) and their relation to apoptosis of pancreatic acinar cells.

METHODSRat models of acute pancreatitis with varied severity were established by retrograde injection of sodium taurocholate at different concentrations via the apancreatobiliary duct. The expressions of Fas and FasL mRNAs and proteins were detected by reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. The apoptosis of acinar cells was quantified by in situ nick ending-labeling technique.

RESULTSof expressions fas, FasL mRNA and protein were colocalized in normal pancreatic acinar cells. In pancreatic tissue with less severe inflammation, the expressions of Fas and FasL mRNA increased significantly, and the apoptosis index of the acinar cells was also elevated; with the increase in the severity of the pancreatitis, the expression of Fas and FasL mRNAs were gradually lowered and apoptosis index of the acinar cells was increased.

CONCLUSIONFas/FasL system might be involved in the renewal and homeostasis of normal pancreatic tissue, and constitute an important pathway mediating the apoptosis of pancreatic acinar cells in AP.

Acute Disease ; Animals ; Apoptosis ; physiology ; Fas Ligand Protein ; biosynthesis ; genetics ; Male ; Pancreas ; pathology ; Pancreatitis ; chemically induced ; metabolism ; pathology ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley ; Taurocholic Acid ; fas Receptor ; biosynthesis ; genetics