miR-208b-3p通过抑制线粒体基因表达加重心力衰竭小鼠能量代谢障碍  

MiR-208b-3p aggravates energy metabolism disorders in mice with heart failure by inhibiting mitochondrial gene expression

作　　者：周双珊 刘源 银萍 石毓君[2] 苏立[1] ZHOU Shuangshan;LIU Yuan;YIN Ping;SHI Yujun;SU Li(Department of Cardiovascular Diseases,the Second Affiliated Hospital of Chongqing Medical University,Chongqing,400014;Department of Pathology,West China Hospital of Sichuan University,Chengdu,Sichuan Province,610041,China)

机构地区：[1]重庆医科大学附属第二医院心血管内科,重庆400014 [2]四川大学华西医院病理研究室,成都610041

出　　处：《陆军军医大学学报》2024年第16期1857-1866,共10页Journal of Army Medical University

基　　金：重庆市自然科学基金面上项目(cstc2021jcyj-msxmX0208)。

摘　　要：目的探究miR-208b-3p对主动脉弓缩窄(transverse aortic constriction,TAC)诱导的心力衰竭小鼠能量代谢的作用及机制。方法将24只小鼠按照随机数字表法分为假手术组(Sham组,n=6)和手术组(n=18)。手术组采用TAC建立心力衰竭模型;假手术组采用TAC相同手术方式,但不结扎主动脉弓。于TAC术后第2周再随机分为Antagomir组(n=6)、Antagomir-NC组(n=6)及TAC组(n=6),Antagomir组、Antagomir-NC组尾静脉分别注射miR-208b-3p antagomir(800μg)及miR-208b-3p antagomir阴性对照(800μg)试剂,每周2次,连续4周。术后第6周行超声心动图评估各组小鼠心功能。HE染色及天狼星红染色观察小鼠心肌组织病理学形态。ATP检测试剂盒检测各组小鼠心肌组织ATP水平。RT-qPCR检测小鼠心肌组织miR-208b-3p、线粒体基因(ND1、ND2、ND3、ND4、ND4L、ND5、ND6、CO1、CO2、CO3、CYTB、ATP6、ATP8)及POLRMT、12SrRNA的表达。双荧光素酶报告基因实验检测miR-208b-3p与潜在靶基因POLRMT的相互结合作用。Western blot检测心肌组织POLRMT及ND1、CO2、CYTB、ATP8蛋白表达水平。结果与Sham组相比,TAC组小鼠心肌miR-208b-3p表达显著增加(P<0.05)。超声检测结果显示,Antagomir组小鼠心脏的射血分数、收缩与舒张功能较TAC组明显改善(P<0.05)。组织切片染色提示,Antagomir组小鼠的心肌细胞肥大、排列紊乱、心肌间质炎细胞浸润等有明显改善(P<0.05)。与TAC组相比,Antagomir组ATP水平明显升高(P<0.05);POLRMT及线粒体基因转录产物(12SrRNA和13个线粒体基因编码多肽)表达量均显著升高(P<0.05),但SDHA、SDHB无变化。双荧光素酶报告基因实验显示miR-208b-3p与POLRMT的CDS区有结合作用。Antagomir组小鼠心肌组织POLRMT、ND1、CO2、CYTB、ATP8蛋白水平表达较TAC组显著增加(P<0.05)。结论miR-208b-3p可通过靶向POLRMT抑制线粒体基因的表达,加剧线粒体能量代谢障碍,并加重心力衰竭小鼠心功能不全及心室重塑。Objective To explore the effect and mechanism of miR-208b-3p on energy metabolism in mice with heart failure(HF)induced by transverse aortic constriction(TAC).Methods Twenty-four mice were randomly divided into sham operation group(Sham group,n=6)and surgery group(n=18).TAC was used to establish an HF model in the surgical group,the sham group received the same surgical procedures as TAC,but no ligation of the transverse aortic arc.At the second week after TAC,the surgery group was randomly divided into Antagomir group(n=6),Antagomir-NC group(n=6)and TAC group(n=6).The mice of the Antagomir group and the Antagomir-NC group were injected with miR-208b-3p antagomir reagent(800μg)and miR-208b-3p antagomir negative control reagent(800μg),respectively by tail vein,twice a week,for 4 consecutive weeks.Echocardiography was performed at the 6th week after surgery to evaluate the cardiac function.HE staining and Sirius red staining were used to observe myocardial histopathology in mice.ATP assay was employed to detect the ATP level in myocardial tissues.RT-qPCR was applied to detect the expression of miR-208b-3p,mitochondrial genes(ND1,ND2,ND3,ND4,ND4L,ND5,ND6,CO1,CO2,CO3,CYTB,ATP6 and ATP8),POLRMT and 12S rRNA in myocardial tissues.Double luciferase reporter assay was conducted to detect the interaction between miR-208b-3p and the potential target gene POLRMT.Western blotting was utilized to detect the changes in the protein levels of POLRMT and ND1,CO2,CYTB and ATP8 in myocardial tissues.Results The expression of miR-208b-3p was significantly higher in myocardial tissues of the TAC group than the Sham group(P<0.05).Echocardiography revealed that the ejection fraction,systolic and diastolic functions were significantly improved in the Antagomir group than the TAC group(P<0.05).Pathological observation showed significantly improved cardiomyocyte hypertrophy,arrangement disorder and myocardial interstititial cell infiltration in the Antagomir group(P<0.05).Compared with the TAC group,the ATP level was significantly incre

关 键 词：miR-208b 心力衰竭 能量代谢 线粒体基因 POLRMT 

分 类 号：R341[医药卫生—基础医学] R394.1R541.602