| ObjectiveHepatic ischemia-reperfusion injury is a pivotal clinical problem occurring in many clinical conditions such as liver cancer,hepatic rupture and liver transplantation.It significantly increases the difficulty and risk of liver surgery.This experiment was established on the basis of the mode of hepatic ischemia-reperfusion in rats which were given the pretreatment of astragaloside ?.By detecting the serum transaminases,inflammatory related factors,and observating the microstructure of liver cells,judgment whether the pretreatment of astragaloside IV had a protective function in hepatic ischemia-reperfusion injury after postoperative at 4 h,8 h,16 h.By using Western Blot and immunohistochemical staining,analysised the expression of autophagy related protein Beclin-1 in liver tissue.The change of autophagosome quantity was observated under the electron microscope in liver cells.Discussion the change of autophagy level and the root of possible mechanisms,to provide the potential theoretical basis on reduceing patients hepatic ischemia-reperfusion injury.MethodsExperimental group:90 pure SD rats,were randomly divided into control group(Sham group),hepatic ischemia-reperfusion group(HIRI group)and the astragaloside IV group(intervention group),30 in each group.According to the postoperative three different time points(4 h,8 h,and 16 h),each time had ten rats.The rats in sham group were not established the model of hepatic ischemia-reperfusion.The rats in HIRI group and intervention group were established the model of hepatic ischemia-reperfusion.The rats in intervention group were given preoperative intraperitoneal injection with astragaloside IV,once a day,7 days in a row.Observation indexes:All serums and liver tissues were obtained at three different time points(4 h,8 h,and 16 h).Serum alanine aminotransferase(ALT)and aspertate aminotransferase(AST)were detected by automatic biochemical analyzer;Serum inflammatory related factors TNF-a and IL-6 were detected by ELISA.The change of liver cells microstructure was observed under the optical microscope.Western Blot and immunohistochemical staining analysised the expression of Beclin-1 in liver tissue.The number of autophagosomes was observed under electron microscope.Results1.The level of serum transaminases:Compared with the Sham group,the level of serum ALT and AST in HIRI group rats at postoperative 4 h,8 h,16 h were significantly higher(P<0.05),reached its zenith at 8 h after reperfusion.Compared with HIRI group,the level of serum ALT and AST in intervention group rats at postoperative 4 h,8 h,16 h were significantly lower(P<0.05).2,The level of serum inflammatory related factors:Compared with the Sham group,the level of serum TNF-a and IL-6 in HIRI group rats at postoperative 4 h,8 h,16 h were significantly higher(P<0.05),reached its zenith at 8 h after reperfusion.Compared with HIRI group,the level of serum TNF-a and IL-6 in intervention group rats at postoperative 4 h,8 h,16 h were significantly lower(P<0.05).3.The pathological features of the liver tissues after H&E staining:The structures of the liver tissues were completely remained ordered in Sham group,whereas a disordered lobular structure,marked hepatocyte necrosis,and polymorphonuclear cell infiltration were observed in HIRI group at 4 h,8 h and 16 h,especially at 8 h.However,compared with HIRI group,intervention group clearly reduced all the pathological features.4.The expression level of autophagy related protein Beclin-1:Compared with the Sham group,the grey value of Beclin-1 in HIRI group rats at postoperative 4 h,8 h,16 h were significantly higher(P<0.05),reached its zenith at 8 h after reperfusion.Compared with HIRI group,the grey value of Beclin-1 in intervention group rats at postoperative 4 h,8 h,16 h were significantly lower(P<0.05).5.Immunohistochemical staining analysis the expression of Beclin-1 in liver tissue:Compared with the Sham group,the average labeling index(LI)in HIRI group rats increased significantly(P<0.05).Compared with HIRI group,the average LI in intervention group rats significantly decreased(P<0.05).6.The change of autophagosome quantity:Compared with the Sham group,the number of autophagosome in HIRI group rats increased significantly(P<0.05).Compared with HIRI group,the number of autophagosome in intervention group rats significantly decreased(P<0.05)ConclusionThe experiment results indicated that pretreatment of astragaloside IV could reduce the levels of aminotransferases and inflammation,reduce the pathological features of liver tissues significantly at the same time.These infered that pretreatment of astragaloside IV could reduce the hepatic ischemia-reperfusion injury in rats.The study also found that pretreatment of astragaloside IV could reduce the expression of autophagy related protein Beclin-1 in levels of protein and organizational,and also reduce the number of autophagosome in liver cells.These showed that the level of autophagy was significantly increased after hepatic ischemia-reperfusion injury,and astragaloside ? may protect hepatic ischemia reperfusion injury in rats in the way of lowering the level of autophagy.The study provided a new theoretical basis on the function of astragaloside ? in reducing hepatic ischemia-reperfusion injury,and had a potential clinical application prospect. |
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