微小RNA-193a-5p对心衰模型大鼠心功能的影响  

Effects of microRNA-193a-5p on cardiac function in rats with heart failure

作　　者：徐泽民 国欣涛 沈娜 方涛 田凤石 Xu Zemin;Guo Xintao;Shen Na;Fang Tao;Tian Fengshi(Department of Cardiac Surgery,Fourth Center Clinical College,Tianjin Medical University,Tianjin 300140,China;Central Laboratory of the Fourth Center Clinical College,Tianjin Medical University,Tianjin 300140,China)

机构地区：[1]天津医科大学第四中心临床学院第四中心医院心外科,300140 [2]天津医科大学第四中心临床学院中心实验室,300140

出　　处：《中华实验外科杂志》2020年第11期2083-2086,共4页Chinese Journal of Experimental Surgery

基　　金：天津市卫生行业重点攻关项目(16kg146)。

摘　　要：目的观察微小RNA(miRNA,miR)-193a-5p靶向调控Ras相关结构域家族蛋白2(RASSF2)表达抑制炎性因子肿瘤坏死因子-α(TNF-α)与白细胞介素(IL)-1β水平及其对心力衰竭模型大鼠心脏功能的影响。方法取60只12~16周龄的清洁级Wistar大鼠(30只雄鼠,30只雌鼠)。2.5%戊巴比妥钠(35mg/kg)麻醉大鼠,暴露大鼠心脏后,随机选择15只大鼠进行缝合(对照组),15只注射磷酸盐缓冲液(PBS)为PBS组,15只注射空白AAV质粒载体为AAV组,15只注射AAV-miR-193a-5p为miR-193a-5p组。PBS组、AAV组和miR-193a-5p组的大鼠冠状动脉左前降支行冠状动脉结扎构建心力衰竭模型。术后4周,彩色多普勒超声心动图检测舒张期和收缩期左心室后壁厚度(LVPWT)、室间隔厚度(IVST)、左心室舒张末期内径(LVEDd)、左心室收缩末期内径(LVESd)和左心室射血分数(LVEF)。应用双荧光素酶报告基因实验探索miR-193a-5p是否与RASSF2的3’非翻译区相互作用。结果术后4周,与对照组比较(4.79±0.33),PBS组(1.57±0.16)和AAV组(1.66±0.18)miR-193a-5p表达降低,而miR-193a-5p组(9.02±0.76)miR-193a-5p表达增高(P<0.05)。与对照组[IVST:(1.88±0.15)mm;LVPWT:(1.93±0.22)mm;LVEDd:(5.44±0.79)mm;LVEF:(52.72±4.98)%]和miR-193a-5p组[IVST:(1.97±0.16)mm;LVPWT:(2.09±0.21)mm;LVEDd:(5.98±0.81)mm;LVEF:(48.83±4.17)%]比较,PBS组[IVST:(2.69±0.28)mm;LVPWT:(2.74±0.36)mm;LVEDd:(6.87±0.84)mm;LVEF:(37.45±3.55)%]和AAV组[IVST:(2.63±0.25)mm;LVPWT:(2.68±0.34)mm;LVEDd:(6.93±0.90)mm;LVEF:(36.10±3.42)%]左心室IVST、LVPWT和LVEDd显著增高,LVEF显著降低(P<0.05),对照组与miR-193a-5p组间差异无统计学意义(P>0.05)。与对照组[TNF-α:(2.01±0.18)μg/L;IL-1β:(8.76±1.48)ng/L;RASSF2:(1.23±0.21)]和miR-193a-5p组[TNF-α:(2.23±0.21)μg/L;IL-1β:(8.90±1.54)ng/L;RASSF2:(1.30±0.25)]比较,PBS组[TNF-α:(5.67±0.52)μg/L;IL-1β:(11.34±2.55)ng/L;RASSF2:(7.44±0.89)]和AAV组[TNF-α:(5.88±0.58)μg/L;IL-1β:(10.99±2.49)ng/L;RASSF2:(7.89±0.97)]血清TNF-α、IL-1β和RASSF2Objective To investigate the role of microRNA(miRNA,miR)-193a-5p in the regulation of Ras-related domain family protein 2(RASSF2)molecule inhibiting the levels of inflammatory factors tumor necrosis factor-α(TNF-α)and IL-1βand their effects on cardiac function in rats with heart failure.Methods Sixty 12-16 week old clean grade Wistar rats(30 males,30 females)were taken.Rats were anesthetized with 2.5%sodium pentobarbital(35 mg/kg).After exposure to rat hearts,15 rats were randomly selected for suture(control group),15 rats were injected with phosphate buffered saline(PBS)as PBS group,15 rats were injected with blank AAV plasmid vector as AAV group,and 15 rats were injected with AAV-miR-193a-5p as miR-193a-5p group.Heart failure model was established in the PBS group,AAV group and miR-193a-5p group.Four weeks after surgery,color Doppler echocardiography was used to detect diastolic and systolic left ventricular posterior wall thickness(LVPWT),interventricular septum thickness(IVST),left ventricular end diastolic diameter(LVEDd),left ventricular end-systolic diameter(LVESd),and left ventricular ejection fraction(LVEF).The dual luciferase reporter gene assay was used to explore whether miR-193a-5p interacts with the 3’untranslated region of RASSF2.Results At 4 weeks postoperatively,compared with the control group(4.79±0.33),the expression of miR-193a-5p was decreased in the PBS group(1.57±0.16)and the AAV group(1.66±0.18),while the expression of miR-193a-5p was increased in the miR-193a-5p group(9.02±0.76,P<0.05).Compared with the control group IVST:[(1.88±0.15)mm;LVPWT:(1.93±0.22)mm;LVEDd:(5.44±0.79)mm;LVEF:(52.72±4.98)%]and the miR-193a-5p group[IVST:(1.97±0.16)mm;LVPWT:(2.09±0.21)mm;LVEDd:(5.98±0.81)mm;LVEF:(48.83±4.17)%],the left ventricle IVST,LVPWT and LVEDd were significantly increased in the PBS group[IVST:(2.69±0.28)mm;LVPWT:(2.74±0.36)mm;LVEDd:(6.87±0.84)mm;LVEF:(37.45±3.55)%]and the AAV group[IVST:(2.63±0.25)mm;LVPWT:(2.68±0.34)mm;LVEDd:(6.93±0.90)mm;LVEF:(36.10±3.42)%],while the

关 键 词：微小RNA 心力衰竭 Ras相关结构域家族蛋白2 炎症 

分 类 号：R541.6[医药卫生—心血管疾病] R-332[医药卫生—内科学]