Study on the mechanism of FTO/DRP1 signaling pathway in arsenic-induced mitochondrial apoptosis in HT22 cells
Objective To explore the potential mechanism of apoptosis of mouse hippocampal neurons(HT22 cells)induced by sodium arsenite(NaAsO2)at the cellular level,and to provide reference for solving neurodegenerative diseases.Methods The effect of NaAsO2 on the viability of HT22 cells was detected by CCK-8 method.HT22 cells were treated with different doses of NaAsO2(0,2,4,6 μmol/L)for 24 hours,the apoptosis rate of HT22 cells was detected by Annexin V-FITC/PI flow cytometry,and the changes of mitochondrial membrane potential of HT22 cells were detected by JC-1 probe and fluorescence microscope.Western blot was used to detect the expression of apoptosis-related proteins Bcl-2,Bax and Cleaved-caspase-3.Real-time fluorescence quantitative PCR and Western blot were used to detect the expression of FTO/DRP1 mRNA and protein.Results Compared with the control group,the viability of HT22 cells treated with different doses of NaAsO2 decreased significantly(F=716.700,P＜0.01),the mitochondrial membrane potential decreased significantly(F=2 028.000,P＜0.01),and the apoptosis rate increased significantly(F=56.030,P＜0.01).The expression of Bcl-2 was down-regulated(F=63.250,P＜0.01),the expression of Bax and Cleaved-caspase-3 was up-regulated(F=39.580,57.480,P＜0.01),and the expression of Bax/Bcl-2 was also up-regulated(F=51.250,P＜0.01).In addition,the expression of FTO,DRP1,p-DRP1(Ser616)and P-DRP1(Ser616)/DRP1 increased significantly(F=62.680,155.000,78.300,18.050,P＜0.01).Conclusion Arsenic may induce apoptosis by activating FTO/DRP1 signal pathway and mitochondrial-mediated apoptosis pathway in HT22 cells.
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