基于SIRT1-FOXO1信号通路探讨安脑平冲方对脑出血大鼠线粒体功能障碍的影响OA

AIM To explore the improvement effects of Annao Pingchong Decoction on neurological function and mitochondrial dysfunction of brain cells in rats with cerebral hemorrhage.METHODS The rat model of cerebral hemorrhage was established by autologous blood injection.The rats were divided into sham operation group,model group,traditional Chinese medicine(TCM)group(7.5 g/kg Annao Pingchong Decoction),western medicine group(5.4 mg/kg edaravone and dexborneol),inhibitor group(10 μg SIRT1 inhibitor EX-527)and inhibitor+TCM group(10 μg EX-527+7.5 g/kg Annao Pingchong Decoction).After 3 days of administration,the neurological function of rats with cerebral hemorrhage was evaluated by mNSS method.Nissl staining was used to observe the pathology of brain tissue.The levels of MDA,8-OHdG and ATP in brain tissue and the activities of mitochondrial respiratory chain complexes(complexes 1,3 and 4)and antioxidant enzymes(SOD,CAT and GPX)were detected by kits.The protein expressions of SOD1,SOD2,CAT,GPX4,SIRT1,p-SIRT1,FOXO1 and Ac-FOXO1 in brain tissues were detected by Western blot.Immunofluorescence was used to detect the fluorescence intensity of SIRT1 and FOXO1 in brain tissue.RESULTS Compared with the sham operation group,the brain tissue around the hemorrhage in the model group was seriously damaged,the levels of oxidative stress(MDA,8-OHdG)increased(P＜0.01),the mitochondrial function(ATP,complex 1,3,4)and the activities of antioxidant enzymes(SOD,CAT,GPX)decreased(P＜0.01),the protein expressions of antioxidant enzymes(SOD1,SOD2,CAT)decreased(P＜0.05),and the protein expressions of SIRT1 and FOXO1 increased(P＜0.05,P＜0.01).Compared with the model group,the TCM group and the western medicine group showed reduced neurological deficit scores(P＜0.01),decreased neuronal damage,and lower levels of MDA and 8-OHdG(P＜0.01).The ATP level and the activities of complexes 1,3,4,SOD,CAT and GPX increased(P＜0.05,P＜0.01),the protein expressions of SOD1,SOD2 and CAT increased(P＜0.05,P＜0.01).The protein expressions of SIRT1,p-SIRT1,and FOXO1 increased(P＜0.05,P＜0.01),while Ac-FOXO1 protein expression was decreased(P＜0.05,P＜0.01).However,there was no significant change in the above indices in the inhibitor group and the inhibitor+TCM group.CONCLUSION Annao Pingchong Decoction can significantly improve neurological deficits in rats with cerebral hemorrhage and alleviate mitochondrial dysfunction and oxidative stress damage in the brain.The mechanism may be related to the regulation of the SIRT1-FOXO1 signaling pathway,thereby promoting the activities and expressions of antioxidant enzymes.