Study On The Mechanism Of Action Of Dahuang Fuzi Jia Wei Decoction On Relieving Acute Pancreatitis

Background:Acute pancreatitis(AP)is an inflammatory response with trypsin abnormal activation in the pancreas,causing pancreatic tissue self-digestion,edema,bleeding and even necrosis.Acute upper abdominal pain,nausea,vomiting,fever and increased blood trypsin are common reactions.It has been divided into mild acute pancreatitis and severe acute pancreatitis(SAP)in clinical.AP is a common clinical acute abdomen and 20%to 30%will develop into SAP.SAP is extremely dangerous,and the pathogenesis is very complex,often accompanied by systemic inflammatory response syndrome(SIRS)and systemic multiple organ dysfunction(MODS),there is no special method for the treatment of this disease at the present stage.Therefore,studying the pathogenesis of AP furtherly and the related regulatory mechanism,seeking new effective diagnostic and therapeutic targets for the disease have become the focus in the present stage.Dahuang Fuzi Decoction is from the golden chamber,it can warm stomach and guide stool.Modern research has shown that Dahuang Fuzi Decoction can inhibit the release of pancreatic enzyme and various cytokines,protect the acinar cells and Chinese medicine commonly used it to treat SAP and other critical disease.This study aims to explore the possible pathogenesis of acute pancreatitis,and providing a new method for treatment of(severe)acute pancreatitis and the new intervention targets.Objective:To investigate the influence of CCN1 on acute pancreatitis mice and its regulation target and explore the protective effect of Dahuang Fuzi Jia Wei Decoction on severe acute pancreatitis mice and its related mechanism.Methods:The mouse model of AP was constructed by intraperitoneal injection of caerulein;The mouse model of SAP was constructed by intraperitoneal injection of caerulein+Lipopolysaccharide(LPS);The mouse model of overexpression of CCN1 Gene was constructed by orbital intravenous injection of adenovirus;We drenched Dahuang Fuzi Jia Wei Decoction to mice;Measured serum amylase value of each group by biochemical approach;Judged the pathological conditions of pancreatic tissue;Tested the expression quantity of CCN1 gene and related inflammatory gene to pancreatic tissue and liver tissue in each mice group;Detected the expression of CCN1 protein in pancreatic tissue and liver tissue of mice by Western Blot;Observed the exudation of neutrophil and macrophage in pancreatic tissue of mice by Immunohistochemistry;Surveyed the expression location of CCN1 protein in pancreatic tissue.Results:1.Serum amylase value in AP group was higher than the control group obviously,the difference between this two groups had statistical significance(P<0.05).Pathological changes of pancreatic tissue(edema,acini necrosis,inflammatory infiltration and vacuolization)in AP group compared with the control group was significantly aggravated,the difference between’ this two groups had statistical significance(P<0.05);Serum amylase value in overexpressed group was lower than the empty gsensor group,the difference between this two groups had statistical significance(P<0.05).Pathological changes of pancreatic tissue(edema,acini necrosis,inflammatory infiltration and vacuolization)in overexpressed group compared with the empty gsensor group was alleviate,the difference between this two groups had statistical significance(P<0.05);The expression quantity of CCN1 gene and CCN1 protein in pancreatic tissue both decreased obviously in AP group compared with the control group and the difference between this two groups had statistical significance(P<0.05);The expression quantity of CCN1 gene of pancreatic tissue and liver tissue in overexpressed group was significantly greater than the empty gsensor group,with the expression quantity of CCN1 protein of pancreatic tissue,the difference was statistically significant(P<0.05);CCN1 protein expressed mainly in intracytoplasmic of pancreatic acinar cells in mice;Compared with the empty gsensor group,reduction inflammation of the pancreas tissue in overexpressed group reflected in the decreased effusion of neutrophils,instead of macrophages.2.Serum amylase value in DHFZD group was similar to SAP group,the result is not statistically significant(P = 0.8486);Pathological changes of pancreatic tissue(edema,acini necrosis,inflammatory infiltration and vacuolization)in DHFZD group compared with the SAP group was alleviate,the difference was statistically significant(P<0.05),but the bleeding was no significant improvement;The expression quantity of CCN1 gene and CCN1 protein in pancreatic tissue both increased obviously in DHFZD group compared with the SAP group and the difference was statistically significant(P<0.05);In addition,the expression of IL-1βmRNA、TNF-a mRNA was descended in DHFZD group with IL-10mRNA increased,compared with the other group(P<0.05);Compared with the SAP group,reduction inflammation of the pancreas tissue in DHFZD group reflected in the decreased effusion of neutrophils,Instead of macrophages.Conclusions:CCN1 could play a protective effect on pancreatic tissue probably via decreasing the inflammatory reaction by reducing neutrophil infiltration in pancreatic tissue of mice.This was expected to provide new view and thinking for clinical treatment of acute pancreatitis.Dahuang Fuzi Jia Wei Decoction could also protect mice through reducing the inflammatory response of pancreatic tissue and the protective effect was probably related with that the Dahuang Fuzi Jia Wei Decoction could increase the expreesion of CCN1 in pancreatic tissue of mice.