有氧运动调节circRNAs m6A甲基化修饰改善高脂饮食小鼠心脏病理性重塑研究  

Aerobic Exercise Improves Cardiac Pathological Remodeling in High-fat Diet Mice by ReguAerobic Exercise Improves Cardiac Pathological Remodeling in High-fat Diet Mice by Regulating m6A Methylation Modified circRNAs lating m6A Methylation Modified

作　　者：徐祖杰 马哲莹 张冰 田振军[3] XU Zujie;MA Zheying;ZHANG Bing;TIAN Zhenjun(College of Physical Education,Taiyuan University of Technology,Taiyuan 030024,China;Division of Sports Science and Physical Education,Tsinghua University,Beijing 100084,China;Institute of Sports and Exercise Biology,School of Physical Education,Shanxi Normal University,Xi’an 710119,China)

机构地区：[1]太原理工大学体育学院,山西太原030024 [2]清华大学体育部,北京100084 [3]陕西师范大学体育学院暨运动生物学研究所,陕西西安710119

出　　处：《体育科学》2024年第2期63-76,共14页China Sport Science

基　　金：太原理工大学高层次引进人才科研启动经费(RY2400000595)。

摘　　要：目的:通过甲基化RNA免疫沉淀测序(MeRIP-seq)和转录组测序(RNA-seq)绘制N6-甲基腺苷(m6A)修饰图谱,探讨环状RNA(circRNAs)m6A甲基化修饰在有氧运动改善高脂饮食小鼠心脏病理性重塑中的作用。方法:C57BL/6J野生型小鼠随机分为正常饮食安静(normal diet-sedentary,ND-SED)组、正常饮食运动(normal diet-exercise,ND-EX)组、高脂饮食安静(high-fat diet-sedentary,HFD-SED)组和高脂饮食运动(high-fat diet-exercise,HFD-EX)组。ND-SED组和ND-EX组小鼠给予普通饲料喂养,HFD-SED组和HFD-EX组小鼠给予60%脂肪供能高脂饲料喂养12周后,ND-EX组和HFD-EX组小鼠进行8周有氧运动。运动干预结束后,检测小鼠体重、体脂含量和心功能。采血、取心脏,检测血清TC和TG含量,酶联免疫吸附测定检测心脏组织TG含量,RT-qPCR检测m6A调节因子,Western blot检测METTL3和FTO蛋白表达,比色法检测心脏组织m6A RNA甲基化含量,MeRIP-seq检测HFD-SED组和HFD-EX组小鼠心脏circRNAs m6A甲基化修饰,SRAMP预测发生m6A甲基化差异和表达差异的circRNA甲基化位点,MeRIP-qPCR验证其m6A水平。结果:8周有氧运动显著降低了高脂饮食小鼠体重和体脂含量,提高了左心室射血分数和短轴缩短率,抑制心脏纤维化和脂质沉积,改善心脏病理性重塑。有氧运动可以显著降低高脂饮食小鼠心脏m6A甲基化水平,甲基转移酶METTL3在其中发挥了重要作用。MeRIP-seq显示,HFD-SED组和HFD-EX组小鼠心脏受m6A修饰的circRNAs大部分都只具有1个m6A修饰位点,且m6A circRNAs主要来自重叠区和外显子。与HFD-SED组比较,HFD-EX组小鼠心脏有13个circRNAs发生了甲基化上调,19个circRNAs发生了甲基化下调,两组差异甲基化的circRNAs大部分来源于重叠区,长度主要富集在1~10000 bps,大部分位于1号、7号和13号染色体上。RNA-seq显示,与HFD-SED组比较,HFD-EX组小鼠心脏有40个circRNAs发生差异表达,其中22个显著上调,18个显著下调。MeRIP-seq和RNA-seObjective:To explore the role of N6-methyladenosine(m6A)methylation modified circular RNAs(circRNAs)in aerobic exercise-induced improvement of cardiac pathological remodeling in high-fat diet mice.Methods:C57BL/6J wild-type mice were randomly divided into four groups:Normal diet-sedentary(ND-SED)group,normal diet-exercise(ND-EX)group,high-fat diet-sedentary(HFD-SED)group and high-fat diet-exercise(HFD-EX)group.The ND-SED and ND-EX groups were fed with normal diet,while the HFD-SED and HFD-EX groups were fed with 60%fat-enriched high-fat diet for 12 weeks,the ND-EX and HFD-EX groups were subjected to 8-week aerobic exercise.After the exercise intervention,the body weight,body fat content,cardiac function,serum TC and TG levelsand heart tissue TG content were measured.m6A regulatory factors were detected by using RT-qPCR.Protein expressions of METTL3 and FTO were determined by using Western blot.m6A RNA methylation content in cardiac tissue was quantified using colorimetric methods.MeRIP-seq was used to detect circRNAs with m6A methylation modification in the heart.SRAMP was employed to predict circRNA methylation sites with m6A methylation difference and expression difference;and MeRIP-qPCR was performed to verify its m6A level.Results:8-week aerobic exercise significantly reduced body weight/fat content of high-fat diet-fed mice,and increased left ventricular ejection fraction(EF)and fractional shortening(FS),inhibited cardiac fibrosis,lipid deposition,and pathological remodeling.Aerobic exercise also decreased m6A methylation level in the heart of high-fat diet-fed mice,while the methyltransferase METTL3 with played an important role in this process.MeRIP-seq analysis revealed that the majority of m6A-modified circRNAs in the mice heart of HFD-SED and HFD-EX groups had only one m6A modification site,with a significant enrichment in overlapping regions and exons.Compared with the HFD-SED group,there were 13 upregulated and 19 downregulated m6A-modified circRNAs in that of HFD-EX group.The differential methylation

关 键 词：环状核糖核酸 有氧运动 N6-甲基腺苷甲基化 肥胖 心脏病理性重塑 

分 类 号：G804.7[文化科学—运动人体科学]