甘草附子汤对顺铂诱导大鼠雪旺细胞线粒体功能和凋亡的影响OA

AIM To explore the effects of Gancao Fuzi Tang on mitochondrial function and apoptosis of RSC96(rat Schwann cells)induced by cisplatin.METHODS RSC96 cells were randomly divided into normal group,model group,aspirin group,Gancao Fuzi Tang group,acrid medicine group and sweet medicine group.The normal group and model group were given 10％blank serum,while each treatment group was given 10％medicine containing serum.After 4 hours of pre-administration,except the normal group,all other groups were interfered with 0.5 μg/mL cisplatin to create a model.After 24 hours,the cell survival rate,apoptosis rate,SOD activity,H2O2 and ATP levels in each group were detected.ELISA was used to detect GSH-Px activity and MDA,Cyt-C,PGC-1α levels.Western blot was used to detect the protein expressions of p-DRP1,DRP1,Bcl-2 and Bax.RT-qPCR was used to detect the mRNA expressions of DRP1,Bcl-2 and Bax.Immunofluorescence was used to detect the MFN1 and MFN2 expressions.RESULTS Compared with the normal group,the survival rate of cells in the model group decreased(P＜0.01),the apoptosis rate increased(P＜0.01),the activities of SOD and GSH-Px,the levels of ATP and PGC-1α decreased(P＜0.01),the levels of MDA and H2O2 increased(P＜0.01),the expressions of p-DRP1/DRP1 and Bax proteins,and DRP1 and Bax mRNA all increased(P＜0.01),and the expressions of Bcl-2 mRNA,and MFN1 and MFN2 proteins fluorescence decreased(P＜0.05,P＜0.01).Compared with the model group,the 24-hour cell survival rate in Gancao Fuzi Tang group increased(P＜0.05),the apoptosis rate decreased(P＜0.01),the activities of SOD and GSH-Px,the levels of ATP and PGC-1α increased(P＜0.05,P＜0.01),and the levels of MDA and H2O2 decreased(P＜0.05,P＜0.01).The expressions of p-DRP1/DRP1,Bax proteins,and DRP1 and Bax mRNA decreased(P＜0.01),while the expressions of Bcl-2 mRNA,and MFN1 and MFN2 proteins fluorescence increased(P＜0.05,P＜0.01).In the sweet medicine group,the apoptosis rate decreased(P＜0.01),GSH-Px activity and ATP level increased(P＜0.01),MDA level decreased(P＜0.01),the expressions of p-DRP1/DRP1,Bax proteins and DRP1 mRNA decreased(P＜0.01),and the expressions of Bcl-2 mRNA and MFN1 protein fluorescence increased(P＜0.01).In the acrid medicine group,GSH-Px activity,ATP level increased(P＜0.01),MDA level decreased(P＜0.01),H2O2 level increased(P＜0.05),PGC-1α level decreased(P＜0.01),and p-DRP1/DRP1 and Bax protein expressions decreased(P＜0.01).CONCLUSION Gancao Fuzi Tang can improve the mitochondrial function and apoptosis of RSC96 cells induced by cisplatin through the compatibility of"Xin"and"Gan",and its mechanism may be related to inhibiting DRP1 phosphorylation to reduce mitochondrial division,and increase the expressions of MFN1 and MFN2 to promote mitochondrial fusion.