miRNA-22在心肌肥厚大鼠中的表达及对PTEN、SERCA的调控作用  

The expression of miRNA-22 in rats with cardiac hypertrophy and its regulatory effects on SERCA,CaN and CaMKⅡ

作　　者：邓丹 DENG Dan(Department of Cardiovascular Diseases,The First Hospital Afflicated to AMU,ChongQing 400000,China)

机构地区：[1]陆军军医大学第一附属医院心血管内科,重庆市400000

出　　处：《河北医药》2021年第6期851-854,859,共5页Hebei Medical Journal

摘　　要：目的探讨miRNA-22在心肌肥厚大鼠中的表达及对SERCA、CaN、CaMKⅡ的影响。方法观察组50只采用腹主动脉狭窄术制备压力超负荷心肌肥厚动物模型的SD大鼠,随机分为A组(20只)、B组(20只)和C组(10只),A组进行microRNA-22腺病毒/空载体腺病毒转染上调miRNA-22表达,B组进行miRNA-22基因敲除下调miRNA-22表达,另选择10只SD大鼠假手术作为对照组。Taqman探针法测定miRNA-22水平,qt-PCR法测定PTEN mRNA、SERCA mRNA水平,Western blot法测定CaN、ANP、SERCA2a、CaMKⅡ蛋白表达水平。结果C组心肌细胞直径增大,全心质量、左心室质量、左心室舒张末期内径(LVDd)、隔舒张末期厚度(IVSTd)、左心室收缩末期左心室内径(LVESD)、左心室后壁舒张末期厚度(LVPWTd)高于对照组(P<0.05),心肌肥厚大鼠造模成功。C组miRNA-22表达高于对照组,PTEN、SERCA表达低于对照组(P<0.05);A组miRNA-22显著上调(P<0.05),PTEN mRNA、SERCA mRNA水平低于C组(P<0.05);B组miRNA-22显著下调(P<0.05),PTEN mRNA、SERCA mRNA水平高于C组(P<0.05);A组、B组、C组3组大鼠心肌细胞CaN、SERCA2a、CaMKⅡ蛋白表达差异有统计学意义(P<0.05),组间两两比较显示,CaN、CaMKⅡ比较A组>C组>B组;SERCA2a比较A组<C组<B组。结论miRNA-22在心肌肥厚中高表达,上调miRNA-22能够增加心肌细胞CaN、CaMKⅡ蛋白表达、降低SERCA2a表达,从而促进心肌肥厚,可能与抑制PTEN、SERCA基因通路有关。Objective To investigate the expression of miRNA-22 in rats with cardiac hypertrophy and its regulatory effects on SERCA,CaN and CaMKⅡ.Methods A total of SD 50 rats with pressure overload myocardial hypertrophy were randomly divided into group A(n=20),group B(n=20)and group C(n=10).The microRNA-22 adenovirus/empty vector adenovirus transfection was performed to up-regulate the expression levels of miRNA-22 in group A,however,the miRNA-22 gene knockdown was performed to down-regulate the expression levels of miRNA-22 in group B.And the other 10 SD rats were enrolled as sham operation group.The expression levels of miRNA-22 were detected by Taqman probe method.The the expression levels of PTEN mRNA and SERCA mRNA were determined by qt-PCR.The expression levels of CaN,SERCA2a and CaMKⅡwere measured by Western Blot.Results The increased diameter of myocardial cells,the total heart mass,left ventricular mass,LVDd,IVSTd,LVESD,LVPWTd in group C were significantly higher than those in control group(P<0.05),which showed that the rats models with cardiac hypertrophy were successfully established.The expression levels of miRNA-22 in group C were significantly higher than those in control group,but the expression levels of PTEN and SERCA were significantly lower than those in control group(P<0.05).The expression levels of miRNA-22 in group A were significantly up regulated(P<0.05),and the expression levels of PTEN mRNA and SERCA in group A were significantly lower than those in group C(P<0.05).Moreover the expression levels of miRNA-22 in group B were significantly down-regulated(P<0.05),and the expression levels of PTEN mRNA and SERCA mRNA in group B were significantly higher than those in group C(P<0.05).In addition there were significant differences in the expression levels of CaN,SERCA2a and CaMKⅡprotein in cardiomyocytes among group A,group B and group C(P<0.05).The comparison between groups each other showed that the levels of CaN and CaMKⅡwere in order group A>group C>group B,and those of SERCA2a were group A<

关 键 词：心肌肥厚 微小RNA-22 肌浆网钙泵 钙调神经磷酸酶 钙调蛋白依赖激酶Ⅱ 

分 类 号：R542.2[医药卫生—心血管疾病]