Objective To investigate the mechanism of dexamethasone in the treatment of acute necrotizing pancreatitis (ANP). Methods The ANP of 48 SD rats were induced by retrograde infusion of sodium taurocholate through biliopancreatic duct.After 30 minutes,the therapy group was administrated with dexamethasone at a dose of 0.2 mg/100 g alone. The control group was administrated with the same amount of 0.9% saline solution.At fourth hour and twelfth hour,8 rats of each group were sacrificed to examine the levels of serum tumor necrosis factor-alpha(TNFα) and serum amylase,to score the degree of pancreatic necrosis and to evaluate acinar cell apoptosis by in situ hybridization by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling(TUNEL). The survial period of 8 rats in each group were observed. Results In therapy group, the level of TNFα was (17.8±2.7) pg/ml and (8.5±1.6) pg/ml,the apoptosis index was （36.94±4.12）% and （ 32.79±3.31）%,the survival period was (33.4±21.5) h.While the control group with the indexes mentioned above were as follows: (53.6±18.7) pg/ml and (37.2±11.1) pg/ml ( P＜0.01),（4.37±1.24）% and （5.12±2.11）% （P＜0.01）,(14.6±5.7) h (P＜0.01) ,the histologic scoring for ANP between therapy group and control group was a significantly distinct (P＜0.01). Conclusion Dexamethasone can induce pancreatic acinar cell apoptosis in this model. Proper leves of TNFα may play an important role in regulating the apoptosis.Apoptosis can protect pancreas from necrosis in ANP.
【Abstract】Objective To study the effects of Chinese traditional medicine Sanqizonggan on bacterial translocation in rats with acute necrotizing pancreatitis (ANP).Methods The rat model of ANP was established by retrograde bilepancreatic duct injection of 5% sodium taurocholate. All rats were randomly divided into three groups: the shamoperation group(n=30), ANP group(n=30), and ANP+Chinese traditional medicine group (n=30). The serum amylase was detected at 0 h,12 h,24 h, and oneweek survival rate and pancreatic histological changes were observed in three groups, and the bacterial translocation from intestinal lumen was examined. Results The survival rate of the group treated with Chinese traditional medicine was significantly higher than that of the ANP group. The rate of bacterial translocation in the treated group significantly decreased. Conclusion The Chinese traditional medicine Sanqizonggan can promote gastrointestinal movement, protect intestinal mucosa and reduce bacterial translocation from intestinal lumen.
Nineteen cats were randomly divided into two groups, 7 cats (group A) recieved about 200 times spotty injections of total of 2 ml of 94% alcohol in pancreatic parenchyma and 12 cats (group B) underwent intraductal alcohoh, partial obstruction of the main pancreatic duct (MPD) and intraparenchymal alcohol. Acute necrotizing pancreatitis occurred in all of the experimental cats after operation. 2 cats in group A (28.6%) died within 48 hours postoperatively. 4 cats in group B (33.3%) died, among them, 3 within 48 hours and 1 died after 2 weeks. Morphological and functional recovery of the exocrine pancreas were found in all the 5 survivals in group A, while 8 cats in group B developed chronic pancreatitis 15 weeks after the operation. The above results show that simple pancreatic necrosis can be recovered after eliminating the etiological factors and if these factors, whatever is primary or secondary still exist and continue to damage the pancreas, chronic pancreatitis may develop.
Objective To explore the influences of hydrogen sulfide （H2S） on acute necrotizing pancreatitis （ANP）. Methods Forty-three SD male rats were grouped by random number table， and divided into five groups：the sham group （n=4）， ANP group 〔n=21， which was divided into 3 subgroups：3， 6， and 12 hours group （n=7）〕， NaCl＋ANP group （n=4）， NaHS＋ANP group （n=7）， and PAG＋ANP group （n=7）. Models of ANP were formed byretrograde cholangiopancreatography injection of 5% sodium taurocholate. The NaCl＋ANP group， NaHS＋ANP group， and PAG＋ANP group rats were given pretreatment of saline， NaHS， or PAG at 1 hour before modelingrespectively. The levels of serum amylase （AMY）， aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， blood urea nitrogen （BUN）， and creatinine （Cr） were detected， and the pathological histological changes of pancreatic tissues were observed. Results The levels of serum AMY， AST， ALT， BUN， and Cr were increased in ANP group. The levels of serum AMY， AST， ALT， BUN， and Cr in the NaHS＋ANP group were higher than those of NaCl＋ANP group （P＜0.05）， and the pathological damage of the pancreatic tissues was more serious in the NaHS＋ANP group. The levels of serum AMY， AST， ALT， BUN， and Cr in the PAG＋ANP group were lower than those of NaCl＋ANPgroup （P＜0.05）， and the pathological damage of pancreatic tissues in the PAG＋ANP group was not so serious as in the NaCl＋ANP group. Conclusions The impairment of liver， kidney， and pancreas function in ANP rats may be related to elevated H2S concentration. Prophylactic administration the PAG of H2S antagonist can improve the function of the liver， kidney， and pancreas， and have the effects of organ protection.
【Abstract】Objective To investigate the preventive role of selective decontamination of the digestive tract (SDD) in gut-originated endotoxemia in acute necrotizing pancreatitis (ANP). Methods A lethal model of ANP was reproduced in Wistar rats by retrograde infusion of artificial bile into the main pancreatic duct. Normal control group (n=6), sham operation group (n=6), ANP group (n=14) and ANP+SDD (polymycin E, tobramycin and nystatin mixture) group (n=8) were randomly devided. Visceral pathologic changes, serum levels of TNFα and IL-1β, intestinal bacterial flora, plasma D(-)lactate and endotoxin contents, as well as the mortality were examined at 72h after operation in each group. Results Necrosis and inflammation of pancreas, with a remarkable elevation of serum TNFα and IL-1β and intestinal flora disturbance (with E.Coli content risen significantly) were seen in ANP rats. Simultaneously, ANP rats displayed elevated plasma concentration of D(-)lactate and endotoxin. In SDD group, enterobacteraceae and yeast were markedly depressed, while anaerobes were well preserved, with the value of B/E 〔Bifidobacterium/E.Coli, log10(CFU/CFU）〕 elevated in the ileac mucous membrane (1.73±1.23 vs -0.37±0.72 in ANP group,P＜0.01) and in the caecum content (∞ vs 0.88±0.77). In addition, depressed levels of D(-)lactate 〔（3.95±1.83） mg/L vs （8.05±3.05） mg/L in ANP group,P＜0.01〕, endotoxin 〔（0.227±0.084） EU/ml vs （0.423±0.155） EU/ml in ANP group, P＜0.01〕 and TNFα 〔（15.41±10.32） ng/L vs （46.79±24.31） ng/L in ANP group P＜0.01〕 in systemic or portal vein were observed in the SDD group. Moreover, SDD group displayed a declined 72h mortality(14.3% vs 58.8% in ANP group, P=0.005). Conclusion ANP is associated with gut barrier disorder and gut flora imbalance, which may exacerbate the process of gut-originated endotoxin translocation. By protecting gut flora and gut barrier against disorder, SDD attenuates ANPrelated endotoxemia and improves the outcome. SDD is advisable for the prophylaxis of gut-originated endotoxemia complicated from ANP.
【Abstract】Objective To investigate therapeutic effect and mechanism of hyperbaric oxygen and ulinastatin respectively or combinatively used to treat acute necrotizing pancreatitis (ANP). Methods One hundred and twenty SD rats were divided into 6 groups randomly: group of normal control, group receiving sham operation, group of untreated acute necrotizing pancreatitis (ANP group), group of acute necrotizing pancreatitis treated with hyperbaric oxygen (HBO group), group of acute necrotizing pancreatitis treated with ulinastatin (ULT group), and group of acute necrotizing pancreatitis treated with combined hyperbaric oxygen and ulinastatin (HBO+ULT group). The rat model of acute necrotizing pancreatitis was established according to Aho HJ et al. Concentrations of amylase, TNFα, TXB2 and 6ketoPGF1α in blood were measured through ELISA or radioimmunoassay. Changes of pancreatic histopathology were investigated. SPSS 10.0 was used in statistical analysis. Results The concentrations of amylase, TNFα, TXB2 in the ANPtreated groups were significantly lower than those of ANP group (P＜0.01) except for 6ketoPGF1α and the levels of amylase and TNFα of HBO group were strikingly higher than those in HBO+ULT group. Only the level of AMS was significantly different between ULT group and HBO+ULT group (P＜0.01). Pancreas histopathological scores(HS) and CD8 counts of ANP group were significantly higher than those the other three group, but CD4 counts and CD4/CD8 ratio were on the contrary (P＜0.05). HS of HBO and ULT were strikingly higher than those of HBO+ULT (P＜0.05).Conclusion ①Hyperbaric oxygen or ulinastatin can effectively decrease the blood levels of enzymes and cytokines and improve the pancreatic immunity. ②Hyperbaric oxygen in combination with ulinastatin are more effective than either of them in the treatment of ANP.
ObjectiveTo investigate the effect of PI3K/Akt/mTOR signaling pathway on liver injury induced by severe acute pancreatitis (SAP). MethodsForty healthy adult male Sprague-Dawley (SD) rats were randomly divided into 4 groups: Sham operation group (SO group), SAP group, PI3K inhibitor LY294002 group (LY294002 group), and mTOR kinase inhibitor rapamycin group (rapamycin group). The rat model with SAP was made by injection with 5% sodium deoxycholate through retrogradely bilio pancreatic duct. Serum levels of amylase (AMY), alanine aminotransferase (ALT), and aspartate transaminase (AST) were detected through the inferior vena at 6 h after modeling. Pathologic change of the liver was observed under the light microscope. TUNEL analysis was used to detect apoptotic index (AI) of the heptocyte. Expressions of Akt, phosphated-Akt (p-Akt), mTOR, phosphated-mTOR (p-mTOR) protein were evaluated by Western blot. Results①Compared with the SO group, the serum levels of AMY, ALT, AST, and the hepatocyte AI were significantly increased among the other three groups (P < 0.05). Compared with the SAP group, the serum levels of AMY, ALT, AST, and the hepatocyte AI were significantly decreased in the LY294002 group and rapamycin group (P < 0.05).②Compared with the SO group, the damages of the liver tissues were aggravated among the other three groups. The pathologies of the liver tissues were ameliorated in the LY294002 group and rapamycin group as compared with the SAP group.③Compared with the SO group, the levels of p-Akt/Akt, p-mTOR/mTOR were significantly increased among the other three groups (P < 0.05). Compared with the SAP group, the levels of p-Akt/Akt, p-mTOR/mTOR were significantly decreased in the LY294002 group (P < 0.05), but in the rapamycin group, only the p-mTOR/mTOR level was significantly decreased (P < 0.05). ConclusionThe activation of PI3K/Akt/mTOR signaling pathway might be one of the reasons for the liver injury induced by SAP and blocking this signaling pathway might be a potential target of preventing progress of SAP and alleviating liver injury induced by SAP.
In the early stage of acute necrotizing pancreatitis (ANP) of experimental monkeys, the concentration of tumor necrosis factor (TNF) in blood was significantly increased. Stilamin could significantly reduce the level of TNF, decrease the mortality, and prolong the survival time of monkeys. The authors draw the conclusion that TNF is an important inflammatory media in the early stage of ANP. Stilamin can significantly inhibit the inflammatory process and has good effect in the treatment of ANP in experimental monkeys.
Objective To study the effect on expression of high mobility group box-1 （HMGB1） mRNA for the expression of zonula occludens-1 （ZO-1） in ileum tissues， and to explore the possible mechanism of intestinal mucosal barrier injury in rats with acute necrotizing pancreatitis （ANP）. Methods Ninety-six male Wistar rats were divided randomly （random number method） into ANP group， ethyl pyruvate （EP）group， and sham operation group. Eight rats of 3 groups were killed to get abdominal aortic blood and ileal tissues at 6， 12， 24， and 48h after operation， respectively.The levels of plasma amylase （AMY） ， D-lactate acid， and the activity of malonyl dialdehyde （MDA） in the ileum tissues were determined by using automatic biochemical analyzer， improved enzymatic spectrophotometry， and thiobarbituric acid （TAB） colorimetry respectively. The pathological changes of ileum tissues were observed under microscopy by HE staining， the expression of ZO-1 protein in ileum tissues was observed by immunohistochemistry （SP method）， and the expressions of HMGB1 mRNA and ZO-1 mRNA in ileum tissues were detected by reverse transcription-polymerase chain reaction （RT-PCR）. Results Compared with ANP group at the same time， levels of AMY， D-lactate acid， and MDA in ileum tissues of EP group were all significantly lower （P＜0.05）. The expression level of HMGB1 mRNA increased at 6 h while ZO-1 mRNA decreased in ANP group. Compared with ANP group at the same time， the expression level of HMGB1 mRNA of EP group was significantly lower while ZO-1 mRNA was higher （P＜0.05）， and the pathological damage in ileum tissues was lighter. Conclusions The decreased expression of ZO-1 in ileum tissues is one of the vitalcauses for intestinal mucosal barrier injury in ANP， and it probably occurs in case of the excessive expression of HMGB1.