三七皂苷R1通过Pink1/Parkin途径调控缺氧/复氧后人心肌细胞线粒体自噬的作用OA

Objective To investigate the mechanism by which Notoginsenoside R1(NGR1)ameliorates hypoxia/reoxygenation(H/R)-induced injury in AC16 human cardiomyocyte cell lines through the regulation of mitophagy.Methods Common genes linked to hypoxia/reoxygenation injury and mitophagy were identified by intersecting data from GeneCards and MitoCarta databases.AC16 cell viability was assessed via CCK-8 assay under varying NGR1 concentrations(0,6.25,12.5,25,50,100,200,300,400,500 μmol/L).AC16 cells were divided into the following groups:control group(Control),model group(H/R),and treatment groups(H/R+NGR1 at 100,200 and 300 μmol/L).Mitochondrial membrane potential(ΔΨm)was measured using 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide(JC-1)staining.Transcriptional levels of mitophagy-related genes(Parkin,Pink1,P62)were quantified by reverse transcription-quantitative PCR(RT-qPCR).Protein ex-pression of mitophagy-related markers(Parkin,Pink1,P62,and LC3BⅡ)was evaluated via Western blot analy-sis.Mitochondrial ultrastructure was visualized by transmission electron microscopy(TEM).Results Compared to the control group,cell viability in the H/R group significantly decreased(P<0.01).Treatment with NGR1 at concentrations above 100 μmol/L significantly enhanced the cell viability of AC16 cells compared to the H/R group(P<0.01).H/R induced a significant decrease in mitochondrial membrane potential(P<0.01),which was re-stored by NGR1 treatment(P<0.01).The mRNA levels of Parkin,Pink1,and P62 in the H/R group were upregu-lated compared to the control group(P<0.05),while NGR1 intervention downregulated their expression(P<0.05).Protein expression levels of Parkin,Pink1,and LC3BⅡ in the H/R group significantly increased,while P62 expression decreased compared to the control group(P<0.01).In contrast,different doses of NGR1 treatment significantly reduced the expression of Parkin,Pink1,and LC3BⅡ while increasing P62 expression(P<0.05).TEM revealed that the mitochondrial structure in the H/R group was severely disrupted,with fragmented and disor-ganized cristae,which was alleviated by NGR1.Conclusion NGR1 ameliorates H/R-induced AC16 cell injury,and its mechanism may be associated with modulating the Pink1/Parkin pathway to suppress excessive mitophagy.