Ginsenoside Rg1 modulates ATP5A1 deacetylation via SIRT3 to attenuate hypoxia/reoxygenation injury in HL-1 cardiomyocytes
This study investigated the mechanism by which ginsenoside Rg1 attenuates hypoxia/reoxygenation(H/R)injury in HL-1 cardiomyocytes by inhibiting the acetylation of ATP synthase subunit alpha(ATP5A1)through silent information regulator 3(SIRT3).In this study,an H/R injury model was constructed by hypoxia for 6 h and reoxygenation for 2 h in HL-1 cardiomyocytes.First,the optimal effective concentration of ginsenoside Rg1 was determined using a cell viability assay kit.Then,lactate dehydrogenase(LDH)leakage was measured using a microplate method to evaluate the protective effect of ginsenoside Rg1 against H/R injury in HL-1 cardiomyocytes.Western blot was further used to detect SIRT3 expression and the acetylation level of mitochondrial proteins in cardiomyocytes.ATP content was measured using a luciferase assay.Immunoprecipitation was used to detect the acetylation level of ATP5A1.The oxygen consumption rate(OCR)was measured using the Seahorse XFp analyzer.Flow cytometry was used to assess cell apoptosis to explore the specific mechanism.The results showed that compared with the control group,the model group had a significant increase in LDH leakage,a decrease in SIRT3 expression,a significant increase in the acetylation levels of mitochondrial proteins and ATP5A1,a decrease in OCR and ATP content,and an increase in cell apoptosis rate.Compared with the model group,the ginsenoside Rg1 group showed a significant decrease in LDH leakage,an increase in SIRT3 expression,a significant decrease in the acetylation levels of mitochondrial proteins and ATP5A1,an increase in OCR and ATP content,and a decrease in cell apoptosis rate.Compared with the ginsenoside Rg1 group,the ginsenoside Rg1+si SIRT3 group showed a significant increase in LDH leakage,a decrease in SIRT3 expression,a significant increase in the acetylation levels of mitochondrial proteins and ATP5A1,a decrease in OCR and ATP content,and an increase in cell apoptosis rate.In conclusion,ginsenoside Rg1 alleviates H/R injury in HL-1 cells by improving mitochondrial respiratory function through SIRT3-mediated inhibition of ATP5A1 acetylation.
ginsenoside Rg1ATP5A1 acetylationhypoxia/reoxygenation injurymitochondrial respiratory function
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