Protective effect of Fructus Mume total flavone against SH-SY5Y cells damage induced by MPP+and its mechanism

Objective:To investigate the neuroprotective effects of Fructus Mume total flavone(FMF)against cell apoptosis and mitochondrial injury induced by 1-methyl-4-phenylpyridinium(MPP^(+))in human neuroblastoma(SH-SY5Y)cells and explore its molecular mechanisms.Methods:MPP^(+) induced SH-SY5Y cells injury model were established in vitro cell culture,the cells were divided into 5 groups:normal control group,model group(250μmol·L^(-1) MPP^(+)),FMF low-and middle-and high-dose experimental group(10,50,100μmol·L^(-1) FMF).After 72 h administration,4’,6-diamidino-2-phenylindole(DAPI)staining was used to observe the effects of different concentrations of FMF on the morphologic changes of apoptotic cells,the ratio of cell apoptosis was measured by Annexin-FITC/PI double staining.The mitochondrial membrane electro-bit were detected by flow cytometry(FCM).The expression of Bcl-2,Bax and Caspase-3 were detected by Western blot.Results:The results of DAPI staining showed that the injury SH-SY5Y cells induced by MPP+were densely condensed,the nucleus showed nuclear shrinkage,showing an apoptotic characteristic morphology;after 72h of FMF action,the apoptotic morphology of the cells showed different degrees of improvement,and the apoptotic number of SH-SY5Y cells also decreased.Compared with that in the normal control group,the apoptotic rate and of mitochondrial membrane electrobit of SH-SY5Y cells in the model group increased significantly(P<0.01),the expression of Bax and Caspase-3 proteins increased significantly(P<0.01),Bcl-2 protein and the ratio of Bcl-2/Bax decreased significantly(P<0.01).Compared with that in the model group,the apoptotic rate and mitochondrial membrane electro-bit of SH-SY5Y cells in FMF groups(10,50,100μmol·L^(-1))were significantly lower,while Bax and Caspase-3 proteins were significantly lower(P<0.01),and Bcl-2 protein and the ratio of Bcl-2/Bax were significantly higher,with statistically significant difference in FMF middle-and high-dose experimental groups(P<0.01).The results indicated that FMF can decrease the experession level of Bax and Caspase-3 and increase the ratio of Bcl-2/Bax,inhibit MPP+induced apoptosis.Conclusion:FMF improves the damage of SH-SY5Y cells induced by MPP+,and plays a neuroprotective effect by regulating the expressions of related proteins in mitochondrial apoptosis pathway.