微小RNA-34a对沉默信息调节因子1的调节作用及该因子对严重烧伤大鼠早期心肌损伤的影响  

Effects of microRNA-34a on regulating silent information regulator 1 and influence of the factor on myocardial damage of rats with severe burns at early stage

作　　者：白晓智 何亭 张聚磊 刘洋 曹梦远 张家宁 蔡维霞 贾艳慧 石继红 苏琳琳 胡大海 

机构地区：[1]空军军医大学西京医院全军烧伤中心,烧伤与皮肤外科,西安710032 [2]空军军医大学西京医院急诊科,西安710032 [3]空军军医大学学员一旅

出　　处：《中华烧伤杂志》2018年第1期21-28,共8页Chinese Journal of Burns

基　　金：国家自然科学基金重点项目（81530064）;国家自然科学基金青年基金项目（81501663、81501666）

摘　　要：目的探讨微小RNA-34a对沉默信息调节因子1（SIRT1）的调节作用及SIRT1对严重烧伤大鼠早期心肌损伤的影响。方法（1）将24只SD大鼠按照随机数字表法（分组方法下同）分为假伤组、单纯烧伤组和SIRT1激动剂组，每组8只。单纯烧伤组、SIRT1激动剂组大鼠背部造成30％体表总面积Ⅲ度烫伤（以下称烧伤），假伤组大鼠背部致假伤。伤后即刻，假伤组、单纯烧伤组大鼠腹腔注射生理盐水50mL／kg，SIRT1激动剂组大鼠腹腔注射生理盐水50mL／kg＋1mg／mL白藜芦醇（50mg／kg）。伤后6h，抽取腹主动脉血制备血清，取心肌组织。（2）取12只SD大鼠乳鼠心肌细胞，分为微小RNA-34a模拟物对照组、微小RNA-34a模拟物组、微小RNA-34a抑制物对照组及微小RNA-34a抑制物组，分别用微小RNA-34a的模拟物对照、模拟物、抑制物对照、抑制物序列进行转染。实时荧光定量反转录-聚合酶链反应（RT-PCR）法检测心肌细胞微小RNA-34a表达，蛋白质印迹法检测心肌细胞SIRT1的蛋白表达。另取心肌细胞，分为微小RNA-34a抑制物对照＋烧伤血清组、微小RNA-34a抑制物＋烧伤血清组及微小RNA-34a抑制物＋烧伤血清＋EX527组。微小RNA-34a抑制物对照＋烧伤血清组心肌细胞转染微小RNA-34a抑制物的对照序列，后2组心肌细胞转染微小RNA-34a抑制物序列。转染48h，微小RNA-34a抑制物＋烧伤血清-4-EX527组心肌细胞用含体积分数10％的单纯烧伤组大鼠血清＋终物质的量浓度为1moL／LEX527的DMEM培养液培养6h，其余2组心肌细胞用含体积分数10％的单纯烧伤组大鼠血清的DMEM培养液培养。蛋白质印迹法检测心肌细胞SIRT1及剪切型半胱氨酸天冬氨酸蛋白酶3、Bax的蛋白表达。（3）取来自（1）的心肌组织，实时荧光定量RT-PCR法检测假伤组和单纯烧伤组大鼠心肌组织中微小RNA-34a及SIRT1的mRNA表达；生物图像导航仪下观察假伤组、单�Objective To explore the effects of microRNA-34a on regulating silent information regulator 1 （SIRT1） and influence of SIRT1 on myocardial damage of rats with severe burns at early stage. Methods （1） Twenty-four Sprague-Dawley （SD） rats were divided into sham injury （SI） group, simple burns （SB） group and SIRT1 agonist （SA） group according to the random number table （the same grouping method below） , with 8 rats in each group. Rats in groups SB and SA were inflicted with 30% total body surface area full-thickness scald （ hereinafter referred to as burns） on the back, and rats in group SI were sham injuried on the back. Immediately after injury, rats in groups SI and SB were intraperitoneally injected with normal saline of 50 mL/kg, and rats in group SA were intraperitoneally injected with normal saline of 50 mL/kg and 1 mg/mL resveratrol of 50 mg/kg. At 6 h post injury, abdominal aortic blood was collected to make serum and myocardial tissue of rats was collected. （2） Myocardial cells of twelve neonatal SD rats were collected and divided into microRNA-34a mimic control （MMC） group, microRNA-34a mimic （MM） group, microRNA-34a inhibitor control （MIC） group, and microRNA-34a inhibitor （MI） group, which were respectively transfected with gene sequences of mimic control, mimic, inhibitor control, and inhibitor of microRNA-34a. The microRNA-34a expression level and protein expression level of SIRT1 in myocardial cells were respectively detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction （RT-PCR） and Western blotting. Another batch of myocardial cells were divided into microRNA-34a inhibitor control ＋ burn serum （MCB） group, microRNA-34a inhibitor ＋ burn serum （MB） group, and microRNA-34a inhibitor ＋ burn serum ＋ EX527 （MBE） group. Myocardial cells in group MCB were transfected with gene sequence of inhibitor control, and myocardial cells in the later groups were transfected with gene sequence of inhib

关 键 词：烧伤 心肌疾病 细胞凋亡 微小RNA-34a 沉默信息调节因子1 炎症反应 

分 类 号：R644[医药卫生—外科学]