人参皂苷Rg_(1)调控SIRT3介导ATP5A1去乙酰化减轻HL-1心肌细胞缺氧/复氧损伤的研究  

Ginsenoside Rg_(1) modulates ATP5A1 deacetylation via SIRT3 to attenuate hypoxia/reoxygenation injury in HL-1 cardiomyocytes

作　　者：陈原原 辛高杰 刘子馨 张会雨 郭帆 胥淑娟 王苑 崔晓珊 郭浩[1] 付建华[1,2] CHEN Yuan-yuan;XIN Gao-jie;LIU Zi-xin;ZHANG Hui-yu;GUO Fan;XU Shu-juan;WANG Yuan;CUI Xiao-shan;GUO Hao;FU Jian-hua(Institute of Basic Medicine,Xiyuan Hospital,China Academy of Chinese Medical Sciences,Beijing 100091,China;National Clinical Research Center for Chinese Medicine Cardiology,Beijing 100091,China)

机构地区：[1]中国中医科学院西苑医院基础医学研究所,北京100091 [2]国家中医心血管病临床医学研究中心,北京100091

出　　处：《中国中药杂志》2024年第19期5273-5280,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金项目(82174219);中国中医科学院科技创新工程项目(CI2021A00912);国家中医心血管病临床医学研究中心专项科研基金项目(CMC2022005)。

摘　　要：探讨人参皂苷Rg_(1)通过沉默信息调节因子3(silent information regulator 3,SIRT3)抑制ATP合成酶α-亚基(ATP synthase subunit alpha,ATP5A1)乙酰化减轻HL-1细胞缺氧/复氧(hypoxia/reoxygenation,H/R)损伤的机制。该研究以HL-1心肌细胞为研究对象,通过缺氧6 h、复氧2 h构建H/R损伤模型。首先采用细胞活力检测试剂盒筛选人参皂苷Rg_(1)的最佳有效浓度,然后通过微量法检测乳酸脱氢酶(lactate dehydrogenase,LDH)漏出量,评估人参皂苷Rg_(1)对HL-1心肌细胞H/R损伤的保护作用。进一步采用蛋白免疫印迹法检测心肌细胞中SIRT3表达量和线粒体蛋白乙酰化水平;荧光素酶法测定三磷酸腺苷(adenosine triphosphate,ATP)含量,免疫共沉淀检测ATP5A1乙酰化水平;Seahorse XFp测定细胞耗氧率(oxygen consumption rate,OCR),流式细胞仪检测细胞凋亡情况。结果显示,与正常组比较,模型组LDH漏出量显著增加,SIRT3表达量降低,线粒体蛋白乙酰化水平和ATP5A1乙酰化水平显著升高,OCR降低,ATP含量减少,细胞凋亡率升高;与模型组比较,人参皂苷Rg_(1)组LDH漏出量显著减少,SIRT3表达量升高,线粒体蛋白乙酰化水平和ATP5A1乙酰化水平显著降低,OCR升高,ATP含量增加,细胞凋亡率减少;与人参皂苷Rg_(1)组比较,人参皂苷Rg_(1)+si SIRT3组LDH漏出量显著增加,SIRT3表达量降低,线粒体蛋白乙酰化水平和ATP5A1乙酰化水平显著升高,OCR降低,ATP含量减少,细胞凋亡率升高。综上所述,人参皂苷Rg_(1)可以通过SIRT3抑制ATP5A1乙酰化改善线粒体呼吸功能减轻HL-1细胞H/R损伤。This study investigated the mechanism by which ginsenoside Rg_(1) 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 Rg_(1) 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 Rg_(1) 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 Rg_(1) 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 Rg_(1) group,the ginsenoside Rg_(1)+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 Rg_(1) alleviates H/R injury in HL-1 cells by i

关 键 词：人参皂苷Rg_(1) ATP5A1乙酰化 缺氧/复氧损伤 线粒体呼吸功能 

分 类 号：R285[医药卫生—中药学]