探讨小续命汤抑制阿尔茨海默病体外细胞模型铁自噬的作用研究OA

Objective:To observe the effects of Xiao-Xu-Ming Decoction(XXMD)on the expression of nuclear receptor coactivator 4(NCOA4)and ferritin heavy chain(FTH1)in glutamate(Glu)-induced neurotoxic injury in mouse hippocampal neuronal cells(HT22),and to explore the mechanism of traditional Chinese medicine in treating Alzheimer's disease(AD).Methods:HT22 cells in the logarithmic growth phase were divided into a control group,a Glu model group,and low-and high-dose XXMD groups.An in vitro AD cell model was established by inducing HT22 cell damage with 20 mmol/L Glu.The cells in the XXMD groups were treated with different concentrations(500 and 1 000 μg/mL)of XXMD.The effects of XXMD on the proliferation and survival rate were detected using the methyl thiazolyl tetrazolium(MTT)assay.Autophagy flux was assessed using mRFP-GFP-LC3 adenovirus transfection.The associations between NCOA4/LC3 and NCOA4/FTH1 expression were examined by immunofluorescence double staining.The protein expression levels of LC3B Ⅱ/Ⅰ,NCOA4,and FTH1 were measured using Western blot analysis.Results:XXMD alleviated Glu-induced damage to HT22 cells in a dose-dependent(P<0.01).Compared with the control group,the Glu group showed increased autophagy flux,elevated protein expression of NCOA4 and LC3B Ⅱ/Ⅰ(P<0.01),and decreased protein expression of FTH1(P<0.01).Compared with the Glu group,the low-and high-dose XXMD groups exhibited reduced autophagy flux,decreased protein expression of NCOA4 and LC3B Ⅱ/Ⅰ(P<0.01),and upregulated FTH1 protein expression(P<0.01).Conclusions:XXMD significantly improves the survival rate of Glu-induced HT22 cells and inhibits abnormal autophagy flux.The mechanism may involve the downregulation of NCOA4 protein expression and upregulation of FTH1 protein expression.