微小RNA-30e在脓毒症急性肺损伤大鼠中的表达及其与炎性因子水平的相关性  

Expression of microRNA-30e in sepsis -induced acute lung injury rats and its correlation with the levels of inflammatory cytokines

作　　者：刘翠[1] 刘舒[2] 黄锦达 陈亦婷[1] 吕娟娟[1] 郑贵浪[1] 曾其毅[1] 

机构地区：[1]南方医科大学珠江医院儿科,广州510282 [2]广东省妇幼保健院产前诊断和遗传病诊疗中心,广州511442

出　　处：《中华实用儿科临床杂志》2016年第6期417-420,共4页Chinese Journal of Applied Clinical Pediatrics

基　　金：国家自然科学基金（81272070）;广东省科技计划项目（2014A020212725）

摘　　要：目的从体内和体外实验2个方面探讨微小RNA-30e（miR-30e）在脓毒症诱导急性肺损伤（ALI）发生时的差异表达及其与炎性因子IL-1β、TNF—α表达的相关性。方法30只雄性SD大鼠随机等量分为5组：正常对照组，脓毒症3、6、12、24h组，腹腔注射脂多糖（LPS）10mg／kg构建脓毒症大鼠ALI模型。体外培养大鼠肺泡巨噬细胞株NR8383，分为空白对照组和以终质量浓度1mg／LLPS刺激3、6、12及24h组，采用反转录聚合酶链反应检测IL-1β、TNF-α及miR-30e表达水平，并观察大鼠肺组织病理改变。结果脓毒症组大鼠肺组织中IL-1β、TNF-α水平均较正常对照组明显上调，差异均有统计学意义（P均〈0．01）；HE染色示脓毒症组大鼠肺组织呈ALl改变；脓毒症组大鼠3、6、12、24h肺组织miR-30e表达水平分别为0．26±0．02、0．41±0．08、0．29±0．05和0．18±0．05，均明显低于正常对照组（1．23±0．24），差异均有统计学意义（P均〈0．01）；NR8383细胞中，与空白对照相比，LPS刺激不同时间点炎性因子IL-1β、TNF—α水平均明显上调，差异均有统计学意义（P均〈0．01）；LPS刺激3、6、12、24h时间点细胞中miR-30e表达水平分别为0．27±0．04、0．55±0．05、0．65±0．02、0．41±0．10，均明显低于空白对照组（1．17±0．21），差异均有统计学意义（P均〈0．01）；各实验组大鼠肺组织中miR-30e的表达与IL-1β、TNF-α水平均呈负相关（IL-1B：r=-0．417，P=0．022；TNF．Or．：r=-0．437，P=0．016）；LPS刺激NR8383细胞后不同时间点细胞中miR-30e的表达与IL-1β、TNF-α水平亦均呈负相关（IL-1β：r=-0．713，P=0．003；TNF-α：r=-0．712，P=0．002）。结论miR-30e在脓毒症大鼠诱导ALI发生时的表达量显著下调，并与炎性因子IL-1β、TNF-α表达水平呈负相关，有望成为脓毒症ALl早期诊断、治疗、预后评估新的生物学标志。Objective To investigate the differential expression of microRNA -30e in sepsis -induced acute lung injury（ALI） and its correlation with intedeukin（IL） - 1β and tumor necrosis factor（TNF） - α from two aspects of in vivo and in vitro. Methods Thirty SD male rats were randomly divided into 5 groups ： normal control group,3 - hour sepsis group ,6- hour sepsis group, 12 -hour sepsis group and 24 -hour sepsis group in equal number. Sepsis -in- duced ALI model was induced by intraperitoneal injection of lipopolysaccharide （LPS, 10 mg/kg）. The rat alveolar mac- rophages NR8383 were divided into blank control group and LPS（ 1 mg/L） stimulated 3,6,12,24 hour groups. Inverse transcription- polymerase chain reaction was used to assay the production changes of IL - 1β, TNF - α and miRNA - 30e in lungs and cells. The injury of lung tissue was evaluated through histopathology. Results The levels of IL - 1βand TNF - α in lung tissues of rats in sepsis groups were obviously up - regulated when compared with those in normal control groups（ all P 〈 0.01 ）. The lung tissue hematoxylin - eosin staining indicated ALI in the sepsis group. The relative expression of miR - 30e in rat lung tissue in sepsis 3,6,12,24 hour groups were respectively 0.26 ± 0.02, 0. 41 ±0.08,0.29 ±0.05 and 0.18 ±0.05 ,which were significantly lower than those in normal control group（ 1.23 ±0. 24,all P 〈0.01 ）. The levels of IL- 113 and TNF -ct in LPS stimulated NR8383 cells at different time points were obviously up - regulated when compared with those in blank control groups （ all P 〈 0.01 ）. The relative expression of miR- 30e in LPS stimulated 3,6,12,24 hour groups were respectively 0.27 ± 0.04,0.55 ±0.05,0.65 ± 0.02 and 0.41 ±0. 10,which were significantly lower than those in blank control group（ 1.17 ±0.21 ,all P 〈0.01 ）. The expres- sion of miR - 30e in lung tissues of groups showed significantly negative correlations with those of IL - 1β and TNF - α （IL-1β：r = -0.417,P=0.022; TNF-α�

关 键 词：微小RNA-30e 脓毒症 肺损伤 急性 NR8383 炎性因子 

分 类 号：R459.7[医药卫生—急诊医学]