circRNA_0000994通过翻译蛋白抑制心肌细胞肥大相关基因表达  

Circular RNA_0000994 inhibits hypertrophy-related gene expression in cardiomyocytes by translating protein

作　　者：郭继深 丰嘉欣 温艺红 陈泽润 李艺 朱杰宁 李晖 徐金东[6] 方咸宏 单志新 GUO Ji-shen;FENG Jia-xin;WEN Yi-hong;CHEN Ze-run;LI Yi;ZHU Jie-ning;LI Hui;XU Jin-dong;FANG Xian-hong;SHAN Zhi-xin(The Second School of Clinical Medicine,Southern Medical University,Guangzhou 510280,China;School of Biology and Biological Engineering,Southern Medical University,Guangzhou 510006,China;School of Medicine,Southern Medical University,Guangzhou 510006,China;Guangdong Provincial Key Laboratory of Clinical Pharmacology,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China;Department of Clinical Laboratory,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China;Department of Anesthesiology,Guangdong Provincial People's Hospital,Guangdong Academy of Medical Sciences,Guangzhou 510080,China)

机构地区：[1]南方医科大学第二临床医学院,广东广州510280 [2]华南理工大学生物科学与工程学院,广东广州510006 [3]华南理工大学医学院,广东广州510006 [4]广东省人民医院(广东省医学科学院)广东省临床药理学重点实验室,广东广州510080 [5]广东省人民医院(广东省医学科学院)检验科,广东广州510080 [6]广东省人民医院(广东省医学科学院)麻醉科,广东广州510080

出　　处：《中国病理生理杂志》2022年第10期1756-1764,共9页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.82070254);广东省自然科学基金资助项目(No.2022A1515012522,No.2022A1515012175,No.2021A1515220122);广州市科技计划项目(No.202102080093);广东省人民医院心血管专项(No.2020XXG003);广东省人民医院国自然培育项目(No.KY0120220028)。

摘　　要：目的:研究环状RNA(circRNA)_0000994对心肌细胞肥大表型的调控作用及机制。方法:通过RT-qPCR检测健康器官捐献者(n=18)与心力衰竭(HF)病人(n=28)心肌组织中circRNA_0000994及其宿主基因溶质载体家族8成员A1(SLC8A1)的表达水平。通过分离和培养新生小鼠心室肌细胞(NMVCs),利用重组腺病毒载体介导circRNA_0000994过表达并检测其对NMVCs肥大表型的影响。采用鬼笔环肽染色检测circRNA_0000994对血管紧张素II(Ang II)诱导的NMVCs肥大反应的影响。人AC16心肌细胞过表达circRNA_0000994后,利用质谱shotgun技术鉴定circRNA_0000994预期翻译蛋白(简称SLC8A1-605aa)的特征性肽段序列。利用NMVCs先后过表达circRNA_0000994及转染siRNA-SLC8A1-605aa,检测SLC8A1-605aa表达对circRNA_0000994抑制NMVCs肥大表型的影响。结果:在HF病人心肌组织中,circRNA_0000994及SLC8A1 mRNA的表达显著增加。过表达circRNA_0000994可降低NMVCs中心脏肥大相关基因肌球蛋白重链7(MYH7)、骨骼肌肌动蛋白α1(ACTA1)和心房钠尿肽(ANP)的表达,抑制Ang II诱导的NMVCs肥大反应(P<0.01)。Western blot及质谱shot-gun分析结果显示,circRNA_0000994可翻译SLC8A1-605aa蛋白。过表达circRNA_0000994和SLC8A1-605aa可一致地抑制NMVCs中心脏肥大相关基因表达,而抑制SLC8A1-605aa表达则可逆转circRNA_0000994对NMVCs肥大表型的抑制作用。结论:circRNA_0000994通过翻译SLC8A1-605aa蛋白发挥抑制心肌细胞肥大的作用。AIM:To investigate the effect of circular RNA(circRNA)_0000994 on cardiomyocyte hypertrophy and the underlying mechanism.METHODS:The expression of circRNA_0000994 and its host gene solute carrier family 8 member 1(SLC8A1)in the myocardium of healthy organ donors(n=18)and patients with heart failure(HF;n=28)was detected by RT-qPCR.The neonatal mouse ventricular myocytes(NMVCs)were isolated and cultured,and the effect of adenovirus-mediated over-expression of circRNA_0000994 on NMVC hypertrophy was investigated.The effect of circRNA_0000994 on angiotensin II(Ang II)-induced NMVC hypertrophy was detected by phalloidin-iFluor 647 staining.The total proteins of human AC16 cardiomyocytes with over-expression of circRNA_0000994 was separated by SDSPAGE,and the protein bands of the predicted SLC8A1-605aa were obtained and used for mass spectrum(MS)shot-gun analysis to identify the characteristic peptides of SLC8A1-605aa.The NMVCs with over-expression of circRNA_0000994were transfected with SLC8A1-605aa siRNA to explore the effect of SLC8A1-605aa on suppression of NMVC hypertrophy by circRNA_0000994.RESULTS:The expression circRNA_0000994 and SLC8A1 mRNA was significantly increased in the myocardium of HF patients.Over-expression of circRNA_0000994 inhibited the expression of cardiac hypertrophy-related genes,including myosin heavy chain 7(MYH7),skeletal muscle actin alpha 1(ACTA1)and atrial natriuretic peptide(ANP),in NMVCs,and suppressed Ang II-induced NMVC hypertrophy(P<0.01).The results of Western blot and MS shot-gun analysis revealed that circ RNA_0000994 could translate SLC8A1-605aa protein.Over-expression of circ RNA_0000994 and SLC8A1-605aa inhibited cardiac hypertrophy-related gene expression in NMVCs,and knock-down of SLC8A1-605aa attenuated the inhibitory effect of circRNA_0000994 on NMVC hypertrophy.CONCLUSION:circRNA_0000994inhibits cardiomyocyte hypertrophy by encoding SLC8A1-605aa protein.

关 键 词：心脏肥大 环状RNA_0000994 溶质载体家族8成员A1 

分 类 号：R541.6[医药卫生—心血管疾病] R363.21[医药卫生—内科学]