微小RNA-155在脓毒症小鼠肝脏内的表达变化及作用研究  

Expression and effects of microRNA-155 in the livers of septic mice

作　　者：王中华[1] 梁艳冰[1] 唐皓[1] 陈志斌[1] 李振宇[1] 吴敬国[1] 杨青[1] 曾丽金[1] 胡旭初[2] 马中富[1] 

机构地区：[1]中山大学附属第一医院普内科,广东广州510080 [2]中山大学中山医学院寄生虫教研室

出　　处：《中国危重病急救医学》2012年第3期154-157,共4页Chinese Critical Care Medicine

基　　金：基金项目：广东省科技计划项目（20098080701070,20098080701004）

摘　　要：目的探讨肝组织中微小RNA-155（miR-155）表达在脓毒症小鼠肝损伤中的作用。方法按随机数字表法将120只BALB／c小鼠分为脓毒症组和正常对照组，每组60只。腹腔注射脂多糖（LPS）20mg／kg复制脓毒症模型，术后0、2、6、12、24、48h每组各取10只小鼠的血及肝组织标本。用酶联免疫吸附试验（ELISA）检测血清和肝组织匀浆中肿瘤坏死因子-α（TNF—α）、白细胞介素（IL-6、IL-10）及血清丙氨酸转氨酶（ALT）含量，并观察肝组织病理改变；用实时荧光定量逆转录-聚合酶链反应（RT—PCR）检测肝组织中miR-155的表达。结果脓毒症组血清和肝组织匀浆中TNF-α、IL-6、IL-10水平均随制模时间延长呈升高趋势，TNF—α、IL-6达高峰后逐渐降低，但仍高于对照组水平；TNF—α（ng／L）于2h达高峰（血清：1538．46±102．12比64．52±18．44，肝：255．26±41．23比60．21±13．55），IL-6（μg／L）于6h达高峰（血清：875．33±102．37比153．72±20．67，肝：9．22±0．82比3．35±0．36），IL-1O（ng／L）于48h达高峰（血清：520．13±88．52比23．43±3．01，肝：260．12±50．38比16．37±3．71），与正常对照组比较差异均有统计学意义（均P〈0．05）。脓毒症组血清ALT活性（U／L）随制模时间延长逐渐增加，至48h达高峰（603．26±70．21比45．84±5．64），与正常对照组比较差异有统计学意义（P〈0．05）。注射LPS后12h，肝组织结构紊乱，大量炎性细胞浸润，肝细胞水肿、坏死；肝组织中miR-155相对表达量在注射LPS后2h开始升高，12h达高峰，为正常对照组的（72．96±9．34）倍（P〈0．05）。结论MiR～155表达增加在脓毒症肝损伤机制中起重要作用。Objective To evaluate the effects of microRNA-155 （miR-155） on liver injury in mice with sepsis. Methods One hundred and twenty BALB/c mice were randomly divided into two groups of equal number according to random number table. Sepsis was induced by intraperitoneal injection of lipopolysaccharide （LPS, 20 mg/kg）. The mice were sacrificed at the time-points of 0, 2, 6, 12, 24, 48 hours. Blood and liver tissue were collected, and the levels of tumor necrosis factor-α （TNF-α）, interleukin （IL-6, IL-10） in serum and liver homogenate and alanine transaminase （ALT） in serum were determined by enzyme linked immunosorbent assay （ELISA）. The injury of liver tissue was evaluated by histopathology. The expression of miR-155 in liver tissue was assessed by fluorescent quantitation reverse transcription-polymerase chain reaction （RT-PCR）. Results The levels of TNF- α, IL-6 and IL-IO in serum and liver homogenate of septic mice increased with passage of time, and then the levels of TNF-α and IL-6 lowered after reaching the peak value, but remained higher than that of control group. TNF-α （ng/L） reached the peak value at 2 hours post-LPS-injection （serum： 1538.46 ± 102.12 vs. 64.52 ± 18.44, liver homogenate： 255.26 ±41.23 vs. 60.21± 13.55, both P〈0.05）. The level of IL-6 （μg/L） reached the peak value at 6 hours post-LPS-injection （serum： 875.33± 102.37 vs. 153.72 ± 20.67, liver homogenate： 9.22 ± 0.82 vs. 3.35 ± 0.36, both P〈0.05）, and that of IL-10 （ng/L） reached the peak value at 48 hours post-LPS-injection （serum： 520.13 ±88.52 vs. 23.43± 3.01, liver homogenate： 260.12 ±50.38 vs. 16.37± 3.71, both P〈0.05）. There were significant differences in above indexes between septic and control group （all P〈0.05 ）. The serum level of ALT （U/L） rose with passage of time, reaching the peak value at 48 hours post-LPS-injection （603.26 ± 70.21 vs. 45.84 ± 5.64, P〈0.05 ）. The values showed significant differences between septic and cont

关 键 词：微小RNA-155 脓毒症 肝损伤 炎症 内毒素 

分 类 号：R459.7[医药卫生—急诊医学]