心肌肥厚大鼠心肌组织及肥大心肌细胞中环状RNA rno_circRNA_016002表达观察  

Expression of circular RNA rno_circRNA_016002 in myocardial tissues and hypertrophic cardiomyo⁃cytes of rats with cardiac hypertrophy

作　　者：邱曼曼 费乔曼 武莉莉 王书影 杨冰[1] 张玲[1] QIU Manman;FEI Qiaoman;WU Lili;WANG Shuying;YANG Bing;ZHANG Ling(College of Basic Medicine,Tianjin Medical University,Tianjin 300070,China;不详)

机构地区：[1]天津医科大学基础医学院,天津300070 [2]中国人民武装警察部队后勤学院

出　　处：《山东医药》2021年第3期23-27,共5页Shandong Medical Journal

基　　金：天津市自然科学基金(19JCQNJC08700);天津市自然科学基金(15JCYBJC25400)。

摘　　要：目的观察心肌肥厚大鼠心肌组织及肥大心肌细胞中环状RNA rno_circRNA_016002的表达变化,并探讨其在心肌肥厚过程中的作用。方法将12只SD大鼠随机分为Control组与异丙肾上腺素(ISO)组,每组6只。ISO组每天腹腔注射ISO 3 mg/kg,Control组注射同等体积的生理盐水,连续注射两周后采用心脏超声检测心肌以明确心肌肥厚大鼠模型建立,然后处死大鼠,取出心脏组织,利用实时定量PCR法检测心肌组织中环状RNA rno_circRNA_016002。将心肌细胞H9C2饥饿过夜后分为ISO 50μmol/L与ISO 100μmol/L组、细胞对照组,ISO 50μmol/L与ISO 100μmol/L组中分别添加50与100μmol/L的ISO进行心肌肥厚的诱导,细胞对照组不作处理,利用实时定量PCR法检测肥大心肌细胞H9C2中环状RNA rno_circRNA_016002;将心肌细胞H9C2分为siRNA+ISO 50μmol/L组、siRNA+ISO 100μmol/L组、siRNA组、阴性对照组,其中siRNA+ISO 50μmol/L与siRNA+ISO 100μmol/L组加入siRNA与转染试剂后,分别再加入50与100μmol/L的ISO处理饥饿过夜;阴性对照组中只加入转染试剂,siRNA组中加入siRNA与转染试剂。加入ISO 48 h后利用实时定量PCR法检测各组心肌细胞H9C2中ANP mRNA与BNP mRNA,采用Western blotting检测各组心肌细胞H9C2中ANP、BNP蛋白质。结果与Control组比较,ISO组环状RNA rno_circRNA_016002的表达水平上调(P<0.01);与细胞对照组比较,ISO 50μmol/L、ISO 100μmol/L组环状RNA rno_circRNA_016002的表达水平均上调(P均<0.01)。siRNA+ISO 50μmol/L与siRNA+ISO 100μmol/L组中ANP、BNP mRNA和蛋白质表达与阴性对照组相比,P均>0.05。结论rno_circRNA_016002在ISO诱导的大鼠肥厚的心肌组织和肥大心肌细胞中表达均上调,并且促进了心肌肥厚。Objective To observe the expression changes of circular RNA rno_circRNA_016002 in the myocardial tissues and hypertrophic cardiomyocytes of rats with cardiac hypertrophy,and to explore its role in the process of cardiac hypertrophy.Methods Twelve SD rats were randomly divided into the Control group and isoproterenol(ISO)group,with 6 rats in each group.The rats in the ISO group were intraperitoneally injected with 3 mg/kg ISO daily,and the Control group with the same volume of normal saline.After two weeks of continuous injection,the rats were subjected to ultrasound examination,and then were sacrificed.Real-time quantitative PCR was used to detect the expression of circular RNA rno_circRNA_016002 in the myocardial tissues.H9C2 cardiomyocytes were starved overnight and were divided into the cell control group,50μmol/L ISO and 100μmol/L ISO groups.Cells in the 50 and 100μmol/L ISO groups were treated with 50 and 100μmol/L ISO,respectively,for cardiac hypertrophy;the cells in the cell control group were not treated.The real-time quantitative PCR was used to detect ANP mRNA and BNP mRNA in H9C2 cardiomyocytes.H9C2 cardiomy⁃ocytes were divided into the negative control group,siRNA group,siRNA+50μmol/L ISO group,and siRNA+100μmol/L ISO group.Only the transfection reagent was added to the negative control group,and siRNA and transfection reagent were added to the siRNA group;the cells in the siRNA+50μmol/L ISO group and siRNA+100μmol/L ISO group were added with siRNA and transfection reagent,and then were treated with 50 and 100μmol/L ISO for overnight starvation.After adding ISO for 48 h,real-time quantitative PCR was used to detect the expression of circular RNA rno_circRNA_016002,ANP mRNA and BNP mRNA in H9C2 cardiomyocytes of each group,and Western blotting was used to detect the expres⁃sion of ANP and BNP proteins in H9C2 cardiomyocytes with siRNA intervention.Results Compared with the control group,the expression level of circular RNA rno_circRNA_016002 in the ISO group was up-regulated(P<0.01);com⁃

关 键 词：环状RNA rno_circRNA_016002 心肌肥厚 

分 类 号：Q331[生物学—遗传学]