Role of autophagy in improving fluoride-induced SH-SY5Y cytotoxicity by grape seed proanthocyanidins
Objective To explore the role of autophagy in improving fluoride-induced SH-SY5Y cyto-toxicity by grape seed proanthocyanidins(GSPE)in human neuroblastoma.Methods SH-SY5Y cells were treated with sodium fluoride(60 mg/L)for 24 h to establish a fluoride-infected cell model,and then were ran-domly divided into the control group,sodium fluoride-infected group,sodium fluoride+low dose GSPE group,sodium fluoride+middle dose GSPE group and sodium fluoride+high dose GSPE group.The cell viability val-ues of each group were detected by CCK-8 assay.Hoechst33342 staining was used to analyze the apoptosis lev-el of each group.Immunofluorescence method was used to analyze the expression of LC3 in each group.The expressions of cleaved-caspase 3,cleaved-PARP,LC3,and p62 were detected by Western blot.Subsequently,Rapamycin was included in the regrouping,and the expressions of cleaved-PARP,LC3,p62 and other proteins in each group were detected,and the changes of apoptosis level in each group were analyzed by Hoechst33342 staining.Results Compared with the control group,the apoptosis level of SH-SY5Y cells in the sodium fluor-ide group was significantly increased(P＜0.05),while the cell viability and autophagy level of SH-SY5Y cells were increased after GSPE intervention(P＜0.05).Compared with the sodium fluoride group,Rapamycin in-tervention could effectively inhibit apoptosis,and the autophagy level was further enhanced in the combination of rapamycin and GSPE together(P＜0.05).Conclusion GSPE can inhibit SH-SY5Y cell apoptosis induced by NaF,and this role may be related to the activation of autophagy.
万方数据
维普
