Guoji laonian yixue zazhi (Jan 2024)
Effect of YTHDF1 on Apoptosis and Oxidative Stress in Myocardial H9c2Cells with Anoxia /Reoxygenation Injury
Abstract
Objective To investigate the effect of YTH domain family protein 1(YTHDF1) on apoptosis and oxidative damage in H9c2 cells under hypoxic reoxygenation(H/R) conditions. Methods The H/R model was constructed with H9c2 cells.Cells were transfected with YTHDF1 small interfering RNA(siRNA) and overexpression plasmid,and cell viability was measured by MTT colorimetric assay.Apoptosis,superoxide dismutase(SOD) activity,malondialdehyde(MDA) content in cells and lactate dehydrogenase(LDH) activity in cell culture medium supernatant were measured by Hirst staining and flow cytometry.The protein levels of YTHDF1,cleaved caspase-3,Bax and FasL in the cells were determined by Western blot,and the apoptosis and oxidative damage of H9c2 cells in the control,H/R,YTHDF1 gene interference and overexpression groups were compared. Results LDH activity and MDA content were significantly increased,SOD activity and cell viability were significantly decreased after H/R injury(P<0.05),and the protein levels of YTHDF1 in H9c2 cells were significantly increased after H/R treatment(P<0.05),YTHDF1 interference alleviated the morphological changes caused by H/R injury,and YTHDF1 overexpression increased the morphological changes caused by H/R injury;cleaved caspase-3,Bax and FasL protein expression was significantly higher after H/R injury treatment than in the control group(P<0.05),cleaved caspase-3,Bax and FasL protein levels were significantly lower in the H/R+pcDNA3.1-YTHDF1 group than in the H/R group(P<0.05) and cleaved caspase-3,Bax and FasL protein levels were significantly higher in the H/R+pcDNA3.1-YTHDF1 group than in the H/R group(P<0.05). Conclusion Downregulation of YTHDF1 inhibited the up-regulation of cleaved caspase-3,Bax and FasL protein levels and decreased oxidative damage in H/R-treated H9c2 cells.
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