体内转染微小RNA-146a对脓毒症小鼠肺的保护作用  

The protective effects of transfected microRNA-146a on mice with sepsis-induced acute lung injury in vivo

作　　者：张建国[1] 丁成志[2] 邵强[2] 刘芬[2] 曾振国[2] 聂成[2] 钱克俭[2] 

机构地区：[1]临沂市人民医院重症医学科,山东临沂276003 [2]南昌大学第一附属医院重症医学科,江西南昌330006

出　　处：《中华危重病急救医学》2015年第7期591-594,共4页Chinese Critical Care Medicine

基　　金：国家自然科学基金（81060152,81101410,81160233）

摘　　要：目的：观察体内转染微小RNA-146a（miR-146a）对脓毒症小鼠肺脏的保护作用。方法选择健康雄性BALB/C小鼠24只，按随机数字表法分为假手术组、脓毒症组、转染组、转染对照组，每组6只。转染组小鼠经气管内预先滴入转染试剂in vivo-jetPEITM处理的miR-146a模拟物（agomir），转染对照组滴入in vivo-jetPEITM处理的阴性对照物。转染12 h后采用盲肠结扎穿孔术（CLP）制备脓毒症模型；假手术组仅开腹游离盲肠，不进行结扎穿刺。各组于术后24 h处死动物，采用实时荧光定量反转录-聚合酶链反应（RT-qPCR）检测肺组织 miR-146a表达，用酶联免疫吸附试验（ELISA）测定支气管肺泡灌洗液（BALF）中肿瘤坏死因子-α（TNF-α）水平，测定肺组织湿/干质量（W/D）比值，光镜下观察肺组织病理学改变并进行肺损伤病理评分。结果脓毒症组、转染组、转染对照组小鼠肺组织miR-146a表达量分别上调至假手术组的（3.56±0.43）、（27.64±3.46）、（3.72±0.54）倍（P＜0.05或P＜0.01），且转染组miR-146a表达上调量明显高于脓毒症组及转染对照组（均P＜0.01）；而脓毒症组与转染对照组差异无统计学意义（P＞0.05）。与假手术组比较，脓毒症组、转染组、转染对照组BALF中TNF-α水平均明显升高（ng/L：511.65±43.47、305.74±34.76、492.27±42.21比50.72±7.23，均P＜0.01），转染组TNF-α水平明显低于脓毒症组及转染对照组（均P＜0.01）。与假手术组比较，脓毒症组、转染组、转染对照组肺W/D比值明显升高（6.11±0.32、5.02±0.29、6.05±0.43比4.18±0.10，均P＜0.01），转染组明显低于脓毒症组及转染对照组（均P＜0.01），且转染组肺损伤病理评分较脓毒症组及转染对照组明显下降（分：6.12±0.75比10.53±1.52、9.73±1.08，均P＜0.01）。结论通过体内转染in vivo-jetPEI^TM荷载的miR-146a agomir可以上调脓毒症小鼠肺组织miR-146a表�ObjectiveTo investigate the protective effect of transfected microRNA-146a （miR-146a） on mice with sepsis-induced acute lung injury （ALI） in vivo.Methods Twenty-four healthy male BALB/C mice were randomly divided into sham group, sepsis group, transfection group and transfection control group, eachn = 6. Mice in transfection group were given miR-146a agomir loaded by in vivo-jetPEITM via airway before reproduction of model, and mice in transfection control group were given negative control loaded by in vivo-jetPEITM only via airway. The septic model was reproduced by cecal ligation and puncture （CLP） 12 hours after transfection , and the mice in the sham group underwent laparotomy and closure only without ligation and puncture of the cecum. The mice of each group were sacrificed at 24 hours post-operation. The expression of miR-146a in lung tissue was determined by real time fluorescence quantitative reverse transcription-polymerase chain reaction （RT-q PCR）, and the quantity of tumor necrosis factor-α （TNF-α） in the bronchial alveolar lavage fluid （BALF） was determined with enzyme-linked immunosorbent assay （ELISA）. The wet/dry ratio of lung （W/D） was determined. The pathohistological changes in the lung were observed and scored. Results The expression of miR-146a showed a significant increase in sepsis group, transfection group and transfection control group, which were （3.56±0.43）, （27.64±3.46） and （3.72±0.54） folds of that in sham group, respectively （〈 0.05 or〈 0.01）. The miR-146a expression in transfection group was significantly increased compared with sepsis group and transfection control group （bothP〈 0.01）, but no statistical difference in the expression was found between sepsis group and transfection control group （P〉 0.05）. Compared with the sham group, higher level of TNF-αin the BALF was found in the sepsis group, transfection group and transfection control group （ng/L： 511.65±43.47, 305.74±34.76, 492.27±42.21 vs. 50.72±7

关 键 词：体内转染 微小RNA-146a 脓毒症 急性肺损伤 

分 类 号：R459.7[医药卫生—急诊医学]