Effect Of Chaihuangqing Yi Huoxue Granule On JNK/P38MAPK Signaling Pathway In Rats With Severe Acute Pancreatitis

objective: To observe the effect of Chaihuangqing Yi Huoxue Granule on JNK/P38 MAPK signaling pathway in rats with severe acute pancreatitis,and to explore its mechanism.Methods:(1)Grouping: 48 adult healthy male SD rats were randomly divided into 3 groups: sham operation group,model group,Chaihuangqing Yi Huoxue Granule group(referred to as Chaihuang group).(2)Modeling: Rats were prepared with 3.5% sodium taurocholate at a dose of 1 ml/kg according to the body weight of the rats.The rat SAP model was prepared by retrograde injection of the pancreaticobiliary tube into the pancreaticobiliary tube.Turn the abdomen after turning the abdominal organs.(3)Dispensing: The Chaihuangqing Yi Huoxue Granule Extract is formulated into a drug solution having a concentration of 0.16 g/ml.(4)Administration: Chaihuangqing Yi Huoxue Granules group was prepared with the solution of Chaihuangqing Yi and Huoxue blood,10ml/kg/time,4 hours apart(0 to 6 o'clock to stop gavage),sham operation group and The model group was orally administered with the same dose of normal saline.(5)Materials: Eight rats were sacrificed at 12 h and 24 h after operation.After gross anatomy,celiac arterial blood and pancreatic tissue were collected and specimens were preserved.(6)Detection: serum amylase and lipase were detected by automatic biochemical analyzer;immunohistochemistry was used to detect the expression of interleukin-1?(IL-1?)and tumor necrosis factor alpha(TNF?)in pancreatic tissue;Western blotting The protein expression of MKK3,MKK4,MKK6,MKK7,JNK and P38 in pancreatic tissue was detected by western blot.The contents of IL-1? mRNA and TNF? mRNA in pancreatic tissue of each group were detected by real-time quantitative PCR.Results:(1)Compared with the sham operation group,the serum AMY and LIP activities of the SAP model group were increased,the difference was statistically significant(p<0.05).Compared with the model group,the activities of AMY and LIP in the serum of the Chaihuang group decreased.Compared with 12 h,the 24 h decline was more pronounced and there was a significant difference(p<0.05).(2)Compared with the sham operation group,the positive amount and score of IL-1? and TNF? in the pancreatic tissue of the model group and the Chaihuang group were increased,the difference was significant(p<0.05);compared with the model group,Chaihuang pancreas activating blood The expressions of IL-1? and TNF? in the immunohistochemistry of pancreatic tissue were significantly decreased in the 12 h and 24 h groups,and the expression was decreased at 24 h compared with 12h(p<0.05).(3)Compared with the sham operation group,the protein expressions of MKK4,MKK7 and JNK in the pancreatic tissue of the model group and the Chaihuang group increased significantly,and the difference was significant(p<0.05).Compared with the model group,the pancreas tissue in the Chaihuang group was compared.The protein expressions of MKK4,MKK7 and JNK were decreased,the difference was statistically significant(p<0.05).(4)Compared with the sham operation group,the protein expression levels of MKK3,MKK6 and P38 in the model group and Chaihuang group were all.The difference was statistically significant(p<0.05).Compared with the SAP model group,the protein expression of MKK3,MKK6 and P38 in the Chaihuang group decreased,and the difference was statistically significant(p<0.05).(5)Compared with the sham operation group,the expressions of IL-1? mRNA and TNF? mRNA in the pancreatic tissue of the model group and the Chaihuang group were significantly increased(p<0.05).Compared with the SAP model group,IL-1? mRNA and TNF? mRNA were compared with the SAP model group.The expression level decreased,and the difference was statistically significant(p<0.05).Conclusion:(1)The amylase and lipase activities in the model group were significantly increased,combined with the previous experimental research,suggesting successful modeling;(2)the expression of inflammatory factors in the model group was significantly increased,and the expression of JNK/P38 MAPK signaling pathway-related protein increased.,suggesting that JNK/P38 MAPK signaling pathway is involved in pancreatic injury of severe acute pancreatitis;(3)Chaihuangqing Yi Huoxue Granule can reduce the expression of inflammatory cytokines in serum and decrease the expression of JNK/P38 MAPK signaling pathway-related proteins,suggesting that Chaihuangqing Yi Huoxue Granules The mechanism of action may be related to the JNK/P38 MAPK signaling pathway.