加味桂枝茯苓汤通过线粒体自噬通路对高糖雪旺细胞损伤的影响OACHSSCD

Objective:To investigate the protective mechanisms of Jiawei Guizhi Fuling Decoction(JGFD)against high glucose-in-duced injury in rat Schwann cells(RSC96)through the PTEN-induced putative kinase 1/Parkin(PINK1/Parkin)signaling path-way.Methods:RSC96 cell injury was induced by high glucose,and 50 mmol/L glucose was selected as the optimal concentration.Cell Counting Kit-8(CCK-8)assay identified 15％JGFD-containing serum as the optimal concentration.Cells were divided into the normal group,model group(50 mmol/L glucose+15％blank serum),JGFD group(50 mmol/L glucose+15％JGFD-containing serum),and siRNA-PINK1 group(siRNA-PINK1 transfection).PINK1 transfection efficiency was verified by Western blot.Mito-chondrial morphology and autophagic structures were observed by transmission electron microscopy.The protein and mRNA expres-sion levels of PINK1,Parkin,P62,Beclin-1,and microtubule-associated protein 1 light chain 3(LC3)were determined by Western blot and real-time quantitative PCR(RT-qPCR),respectively.Reactive oxygen species(ROS)levels were measured by flow cytom-etry.Results:High glucose significantly injured RSC96 cells,and 50 mmol/L glucose resulted in relatively higher cell viability.The protective effect of 15％JGFD-containing serum was optimal.Compared with the normal group,ROS levels were significantly in-creased in the model group(P＜0.01),whereas the expression levels of PINK1,Parkin,Beclin-1,and LC3 were significantly de-creased and P62 expression was significantly increased(P＜0.01).Severe cellular damage and the absence of autophagic structures were also observed.Compared with the model group,the JGFD group exhibited significantly reduced ROS levels(P＜0.01),rever-sal of the above molecular expression changes(P＜0.05 or P＜0.01),alleviated cellular damage,and restoration of autophagic structures.In contrast,the siRNA-PINK1 group reversed the protective effects of JGFD(P＜0.01),accompanied by aggravated cel-lular damage and loss of autophagic structures.Conclusion:JGFD exerts a protective effect against RSC96 cell injury by alleviating cellular damage through PINK1/Parkin signaling pathway-mediated mitophagy.