丙酮酸激酶同工酶M2介导长非编码RNA RP11-879F14.2发挥抑制心肌细胞肥大作用  

Long Non-coding RNA RP11-879F14.2 Inhibits Cardiomyocyte Hypertrophy via Upregulating PKM2 Expression

作　　者：李艺 温艺红 伍华燕 姜佳雪 欧涛 陈凯茵 刘宇鹏[3] 单志新[1] LI Yi;WEN Yi-Hong;WU Hua-Yan;JIANG Jia-Xue;OU Tao;CHEN Kai-Yin;LIU Yu-Peng;SHAN Zhi-Xin(Guangdong Provincial Key Laboratory of Clinical Pharmacology,Medical Research Institute,Guangdong Provincial People’s Hospital,Guangdong Academy of Medical Sciences,Southern Medical University,Guangzhou 510280,China;School of Medicine,South China University of Technology,Guangzhou 510006,China;Department of Cardiology,Guangdong Provincial People’s Hospital,Guangdong Academy of Medical Sciences,Southern Medical University,Guangzhou 510080,China)

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

出　　处：《中国生物化学与分子生物学报》2024年第4期544-553,共10页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金资助项目(No.82070254,82200325,82300277);广东省自然科学基金项目(No.2022A1515012522,No.2022A1515012175,No.2021A1515220122,No.2023A1515010201,No.2021A1515111173);广州市基础与应用基础研究项目(No.202201011627);广东省人民医院国自然培育项目(No.KY0120220028)资助

摘　　要：越来越多的证据表明,长非编码RNA(lncRNA)在生物学功能调节中发挥着重要的作用。实时定量PCR(RT-qPCR)检测显示,与健康器官捐献者(n=23)相比,长非编码RNARP11-879F14.2在心力衰竭(heart failure,HF)患者(n=21)心肌中表达水平显著升高,但其在心肌肥厚调节中可能的作用和机制尚不清楚。利用腺病毒介导在乳小鼠心肌细胞(neonatal mouse ventricular cardiomyocytes,NMVCs)和乳大鼠心肌细胞(neonatal rat ventricular cardiomyocytes,NRVCs)中过表达RP11-879F14.2,检测对NMVCs中心肌肥厚相关基因,包括肌球蛋白重链(MYH7)、骨骼肌肌动蛋白(Acta1)以及心钠素(NPPA)表达的影响,结果显示,过表达RP11-879F14.2可显著抑制NMVCs和NRVCs中心肌肥厚相关基因表达。RT-qPCR和Western印迹结果证实,当过表达RP11-879F14.2时可显著促进NMVCs中丙酮酸激酶同工酶M2(PKM 2)表达。并且,过表达PKM 2和RP11-879F14.2可一致地抑制NMVCs和NRVCs中心肌肥厚相关基因表达和去氧肾上腺素(phenylephrine,PE)诱导NRVCs的表面积增加,而敲降PKM 2可逆转RP11-879F14.2对NMVCs中心肌肥厚相关基因表达的抑制作用。利用Seahorse 96XF细胞能量分析仪检测显示,过表达RP11-879F14.2和PKM 2可一致增加NMVCs中葡萄糖代谢水平,而沉默PKM 2对RP11-879F14.2上调NMVCs中糖酵解、三羧酸(TCA)循环和线粒体电子传递链(ETC)相关基因的表达有显著的抑制作用。因此,PKM 2介导RP11-879F14.2发挥抑制心肌细胞肥大的作用。The increasing evidence suggests that long noncoding RNAs play an important role in regulating biological functions.Comparing with the healthy organ donors(n=23),the results of real-time quantitative polymerase chain reaction(RT-qPCR)assay showed that long noncoding RNA RP11-879F14.2 was significantly increased in the myocardium of patients with heart failure(HF)(n=21),however,the role and mechanism of RP11-879F14.2 in cardiac hypertrophy remains unclear.The effect of adenovirus-mediated overexpression of RP11-879F14.2 on the expression of hypertrophy-related genes,including myosin heavy chain 7(MYH 7),skeletal muscle actin alpha 1(ACTA 1)and natriuretic peptide type A(NPPA),was evaluated,and the RT-qPCR results revealed that overexpression of RP11-879F14.2 could markedly inhibit the expression of cardiac hypertrophy-related genes in neonatal mouse ventricular cardiomyocytes(NMVCs)and neonatal rat ventricular cardiomyocytes(NRVCs).The results of RT-qPCR and Western blotting showed that RP11-879F14.2 could efficiently enhance the expression of pyruvate kinase M2(PKM2)in NMVCs.Overexpression of PKM 2 and RP11-879F14.2 could consistently attenuate the hypertrophy-related genes expression in NMVCs and NRVCs,and inhibited the increase of cell size of phenylephrine(PE)-induced NRVCs.Moreover,knock-down of PKM 2 could reverse the inhibitory effect of RP11-879F14.2 on the cardiac hypertrophy-related genes expression in NMVCs.The glucose metabolic alterations were accessed by using Seahorse XF96 extracellular flux analyzer.Overexpression of RP11-879F14.2 and PKM 2 could consistently enhance glucose metabolism in NMVCs,but knock-down of PKM 2 could inhibit some RP11-879F14.2-promoted glycolysis-related genes,TCA cycle-related genes and mitochondrial ETC-related genes expression in NMVCs.Therefore,RP11-879F14.2 inhibits cardiomyocyte hypertrophy via upregulating PKM 2 expression.

关 键 词：心肌肥厚 心肌细胞 长非编码RNA 丙酮酸激酶同工酶M2 

分 类 号：Q52[生物学—生物化学]