The Research On The Mechanism And Observation On Therapeutic Effect Of Selective Inducing The Apoptosis Of SIRS Relevant Cells To Attenuate Severe Acute Pancreatitis

Objective:1.To explore whether 5-FU in regional arterial infusion can selectively induce apoptosis of SIRS-dominated cells: pancreatic acinar cells,PMN in blood,alveolar monocyte macrophages,and inhibit the necrosis of the above cells,thus in SAP-SIRS the source and development process interrupt this inflammatory cascade to achieve the purpose of treating SAP.2.To explore the role of apoptosis regulatory genes(Bax,Bcl-2,P53)in apoptosis.Methods:Twenty-four healthy SD rats were retrogradely pumped into the pancreaticobiliary duct by 5% sodium taurocholate(0.1ml/100g)to establish a severe acute pancreatitis(SAP)model,followed by catheterization of the left gastric artery.After successful modeling,they were randomly divided into three groups of A,B,and C,with 8 in each group.In Group A,physiological saline(40 mg/kg)was pumped through an arterial catheter at a rate of 6ml/h for 6h,Group B was pumped 5-FU(40 mg/kg)via the tail vein at a rate of6ml/h for 6h,Group C was pumped 5-FU(40 mg/kg)via an arterial catheter at a rate of 6ml/h for 6h.The survival of the rats was observed 6 hours after the administration,and the levels of amylase(AMS),lipase(LPS)and inflammatory cytokines(IL-1,IL-6,TNF-α)in the serum were measured after the rats were sacrificed.The pathological changes in the pancreas tissues were observed.The apoptotic morphology of serum PMN and alveolar mononuclear macrophage was observed by confocal laser scanning micro scope.Tunel method to detect apoptosis index of pancreatic acinar cells in rats.Flow cytometry to detect blood PMN and Alveolar monocyte-macrophage apoptosis rate.Western blot was used to detect the expression of P53,Bax,and Bcl-2 proteins in the apoptotic pathway of pancreatic tissue.Results:There was no statistical significance in the survival of the three groups of rats(P > 0.05).Compared with Group A,the blood amylase and lipase concentrations in Group B and Group C were significantly reduced,and the expression levels of IL-1,IL-6 and TNF-α in serum were significantly decr eased(P<0.05);The degree of pancreatic histopathology was reduced;Blood PMN and alveolar monocytes-macrophages showed obvious signs of apoptosis,and the apoptosis rate was significantly increased by flow cytometry(P < 0.05);The expression level of pancreatic tissue-related apoptotic protein P53 increased,the expression of proapoptotic protein Bax increased,and the expression of anti-apoptotic protein Bcl-2 decreased(P<0.05);However,when the apoptotic index of pancreatic acinar cells was measured by the Tunel method,there was no statistically significant difference between Group B and Group A(P=0.08),Compared with Group A and Group B,the apoptosis index of Group C increased significantly(P <0.05).Conclusion:1.Regional arterial infusion of 5-FU may be through the selective induction of apoptosis of SIRS-leading cells: pancreatic acinar cells,blood PMN,alveolar monocytes and macrophages,and inhibit the necrosis of these cells.The origin and development process of SAP-SIRS interrupts this cascade of inflammation to achieve the purpose of treating SAP.2.The apoptosis regulatory gene P53 in this experiment may play an apoptosis regulating role mainly by regulating the ratio of Bax / Bcl-2 in the mitochondrial pathway.