长链非编码RNA HOTAIR靶向miR-138对脓毒症诱导的大鼠心肌炎症反应和氧化应激的影响及作用机制研究  

Effect of long non-coding RNA HOTAIR targeting miR-138 on sepsis-induced myocardial inflammation and oxidative stress in rats and its mechanism

作　　者：范腾阳 高冬梅 喻荣华[3] 柯迪 肖雪[1] FAN Teng-Yang;GAO Dong-Mei;YU Rong-Hua;KE Di;XIAO Xue(Department of General Medicine,the Affiliated Hospital of Zunyi Medical College,Zunyi 563000,China)

机构地区：[1]遵义医学院附属医院全科医学科,遵义563000 [2]遵义市红花岗区妇幼保健计划生育服务中心,遵义563000 [3]遵义医学院附属医院核医学科,遵义563000

出　　处：《中国免疫学杂志》2020年第2期146-153,共8页Chinese Journal of Immunology

基　　金：贵州省科学技术基金项目(黔科合基础[2018]1197)

摘　　要：目的:探究同源盒基因转录反义RNA(HOTAIR)与miR-138对脓毒症诱导的大鼠心肌炎症反应和氧化应激的影响及作用机制。方法:将大鼠分为Control组、CLP组、siRNA+CLP组、si-HOTAIR+CLP组,构建大鼠CLP模型,PanoView b1500检测大鼠左心室收缩压(LVSP)和左心室舒张末压(LVEDP),RT-PCR检测HOTAIR和miR-138表达水平,HE染色检测心肌病理损伤,试剂盒检测血清炎症因子TNF-α、IL-1β、IL-6、心肌钙蛋白Ⅰ(cTnⅠ)、肌酸激酶同工酶(CK-MB)、丙二醛(MDA)和超氧化物歧化酶(SOD)的水平,Western blot检测NF-κB信号通路蛋白表达水平;将心肌细胞分为Control组、LPS组、siRNA+LPS组、si-HOTAIR+LPS组、si-HOTAIR+LPS+inhibitor组,RT-PCR检测HOTAIR和miR-138表达水平,荧光素酶报告检测HOTAIR与miR-138靶向关系,MTT检测细胞活性,Hoechst检测细胞凋亡,试剂盒检测MDA和SOD的水平,Western blot检测NF-κB信号通路蛋白表达水平。结果:在动物实验中,与Control组比较,CLP组LVSP、细胞活性、miR-138、SOD水平显著降低,LVEDP、病理性改变、细胞凋亡率、HOTAIR、cTnⅠ、CK-MB、TNF-α、IL-1β、IL-6、MDA及NF-κB通路蛋白表达水平显著升高;与CLP组比较,si-HOTAIR+CLP组LVSP、细胞活性、miR-138、SOD水平显著升高,LVEDP、病理性改变、细胞凋亡率、HOTAIR、cTnⅠ、CK-MB、TNF-α、IL-1β、IL-6、MDA及NF-κB通路蛋白表达水平显著降低。在细胞实验中,与Control组比较,LPS组细胞活性、miR-138、SOD水平显著降低,细胞凋亡率、HOTAIR、MDA、NF-κB通路蛋白表达水平显著升高;与LPS组比较,si-HOTAIR+LPS组细胞活性、miR-138、SOD水平显著升高,细胞凋亡率、HOTAIR、MDA、NF-κB通路蛋白显著降低;与si-HOTAIR+LPS组比较,si-HOTAIR+LPS+inhibitor组细胞活性、SOD水平降低,细胞凋亡率、MDA、NF-κB通路蛋白表达水平升高。在荧光素酶报告实验中,与HOTAIR wt组比较,HOTAIR wt+miR-138 mimic组荧光素酶活性显著降低。结论:HOTAIR可以通过沉默mObjective:To investigate HOX transcript antisense RNA(HOTAIR)targeting MicroRNA-138(miR-138)on sepsis-induced myocardial inflammation and oxidative stress in rats and its mechanism.Methods:Rats were divided into control,CLP,siRNA+CLP,si-HOTAIR+CLP group,Rat model of sepsis was established by CLP,LVSP and LVEDP of rats were detected by PanoView b1500,RT-PCR was performed for measuring the levels of HOTAIR,miR-138,myocardial pathological injury were observed by HE staining,the levels of TNF-α,IL-1β,IL-6,cardiac troponinⅠ(cTnⅠ),creatine kinase-MB(CK-MB),Malondialdehyde(MDA)and Superoxide Dismutase(SOD)were measured by kits,Western blot was used to determine the protein levels of NF-κB signal pathway;cardiomyocytes were divided into Control,LPS,siRNA+LPS,si-HOTAIR+LPS,si-HOTAIR+LPS+inhibitor group,RT-PCR was performed for measuring the levels of HOTAIR,miR-138,the luciferase reporter assay was performed for measuring the relationship between miR-138 and HOTAIR,cell viability was measured by MTT,cell apoptosis was determined by Hoechst,MDA and SOD level were measured by kits,Western blot was used to determine the protein levels of NF-κB signal pathway.Results:In animal experiments,compared with Control group,the LVSP,cell viability,miR-138,SOD levels of CLP group decreased significantly,and the LVEDP,myocardial histopathological injury,cell apoptosis rate,HOTAIR,cTnⅠ,CK-MB,TNF-α,IL-1β,IL-6,MDA and protein of NF-κB signal pathway levels increased significantly.Compared with CLP group,the LVSP,cell viability,miR-138,SOD levels of si-HOTAIR+CLP group increased significantly,and the LVEDP,myocardial histopathological injury,cell apoptosis rate,HOTAIR,cTnⅠ,CK-MB,TNF-α,IL-1β,IL-6,MDA and protein of NF-κB signal pathway levels decreased significantly.In cytological experiments,compared with Control group,the cell viability,miR-138,SOD levels of LPS group decreased significantly,and the cell apoptosis rate,HOTAIR,MDA and protein of NF-κB signal pathway levels increased significantly.Compared with LPS group,

关 键 词：脓毒症 非编码RNA HOTAIR miR-138 

分 类 号：R542.2[医药卫生—心血管疾病]