α-酮戊二酸介导异柠檬酸脱氢酶3A发挥抑制心肌细胞肥大的作用  

IDH 3 A Inhibits Cardiomyocyte Hypertrophy via Elevatingα-Ketoglutarate Level

作　　者：伍华燕 温艺红 赵亨利 高原[2] 周川孟 王娅 朱杰宁 单志新 WU Huayan;WEN Yihong;ZHAO Hengli;GAO Yuan;ZHOU Chuanmeng;WANG Ya;ZHU Jiening;SHAN Zhixin(School of Medicine,South China University of Technology,Guangzhou 510006,China;Guangdong Provincial People’s Hospital,Guangdong Academy of Medical Sciences,Southern Medical University,Guangzhou 510080,China;Guangdong Provincial Key Laboratory of Clinical Pharmacology,Guangdong Provincial People’s Hospital,Guangdong Academy of Medical Sciences,Southern Medical University,Guangzhou 510080,China)

机构地区：[1]华南理工大学医学院,广东广州510006 [2]南方医科大学附属广东省人民医院//广东省医学科学院,广东广州510080 [3]广东省临床药理学重点实验室//南方医科大学附属广东省人民医院//广东省医学科学院,广东广州510080

出　　处：《中山大学学报(医学科学版)》2025年第2期275-283,共9页Journal of Sun Yat-Sen University:Medical Sciences

基　　金：国家自然科学基金(82470253);广东省自然科学基金(2022A1515012522,2022A1515012175)。

摘　　要：【目的】探究异柠檬酸脱氢酶3A(IDH3A)对心肌细胞肥大的调控作用和潜在机制。【方法】通过实时荧光定量PCR(RT-qPCR)和Western blot实验检测健康志愿者(n=10)与心力衰竭患者(n=10)心肌组织标本、假手术组与主动脉弓缩窄术(TAC)小鼠心肌标本以及去氧肾上腺素(PE)诱导的乳大鼠心肌细胞(NRVCs)中IDH3A的表达水平。利用腺病毒介导在PE诱导的NRVCs中过表达IDH3A,检测其对NRVCs肥厚相关基因表达的影响,并通过鬼笔环肽染色检测IDH3A对NRVCs面积的影响。利用序列突变方式构建消除IDH3A酶活性的突变体(IDH3A_D208A),通过检测其对NRVCs肥大表型、ATP和ROS水平影响来验证IDH3A是否依赖酶活性发挥调节心肌细胞肥大作用。Western blot和鬼笔环肽染色检测外源给予α-酮戊二酸(AKG)对心肌细胞肥大的影响。【结果】IDH3A在心衰患者心肌、TAC手术小鼠心肌、PE诱导的NRVCs中表达均显著下调(分别为P=0.0052、P=0.0266、P=0.0413和P=0.0066)。过表达IDH3A可显著抑制PE诱导的NRVCs中肥厚相关基因表达和细胞表面积增大(分别为P<0.0001、P=0.0001和P=0.0002)。ATP和ROS水平检测结果表明过表达IDH3A能够抑制PE诱导的NRVCs中ATP和ROS水平的增加(分别为P=0.0012和P<0.0001),而无酶活性的IDH3A突变体则不具备该作用。外源给予AKG可以,而无酶活性的IDH3A突变体无法抑制PE诱导的NRVCs肥大。【结论】IDH3A通过生成AKG发挥抑制心肌细胞肥大的作用,为以IDH3A为靶点的心肌肥厚治疗研究提供科学资料。【Objective】To investigate the regulatory effect and potential mechanisms of isocitrate dehydrogenase 3A(IDH3A)on cardiomyocyte hypertrophy.【Methods】The expression of IDH3A in the myocardium of healthy volunteers(n=10)and patients with heart failure(HF)(n=10),and in the myocardium of mice subjected to transverse aortic constriction(TAC)surgery and sham operation,as well as in phenylephrine(PE)-induced neonatal rat ventricular cardiomyocytes(NRVCs),was assessed by real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot assay.The effect of adenovirus-mediated overexpression of IDH 3A on the expression of hypertrophy-related genes in PE-induced NRVCs was also evaluated.The effect of IDH 3A on NRVCs area was examined by phalloidin staining assay.A mutant of IDH 3A with abolished enzymatic activity,IDH 3A_D 208 A,was generated through site-directed mutagenesis.The impact of this IDH 3A mutant on the hypertrophic phenotype,ATP and ROS levels in NRVCs was evaluated to investigate whether the regulatory role of IDH 3A in cardiomyocyte hypertrophy was dependent on its enzymatic activity.The effect of exogenousα-ketoglutaric acid(AKG)on cardiomyocyte hypertrophy was also detected by Western blot and phalloidin staining assay,respectively.【Results】IDH 3A was significantly decreased in the myocardium of HF patients,in the myocardium of TAC-operated mice,and in PE-induced NRVCs(P=0.0052,P=0.0266,P=0.0413 and P=0.0066,respectively).Overexpression of IDH 3A markedly suppressed the expression of hypertrophy-related genes and the increase of cell size of PE-induced NRVCs(P<0.0001,P=0.0001 and P=0.0002,respectively).The ATP and ROS analysis indicated that IDH 3A inhibited the increases of ATP and ROS levels in PE-induced NRVCs(P=0.0012 and P<0.0001,respectively),whereas the enzymatically inactive IDH 3A mutant lacked this effect.Exogenous AKG provision could,but overexpression of IDH 3A mutant failed to suppress PE-induced NRVCs hypertrophy.【Conclusion】IDH 3A inhibits cardiomyocyte hypertrophy via

关 键 词：心肌肥厚 异柠檬酸脱氢酶3A Α-酮戊二酸 心肌细胞 线粒体 

分 类 号：R363.2[医药卫生—病理学]