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

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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-1MPP+), FMF low- and middle- and high-dose experimental group (10,50,100 μmol·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 electro-bit 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.
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