下调lncRNA Neat1表达通过减少NF-κB P65的细胞核转位保护大鼠心肌缺血再灌注损伤  

Down-regulating lncRNA Neat1 expression protecting myocardial ischemiareperfusion injury in rats by reducing nuclear translocation of NF-κB P65

作　　者：崔勤涛[1] 王学惠[2] 刘永强[1] 刘晓晨[1] 段长恩 CUI Qintao;WANG Xuehui;LIU Yongqiang;LIU Xiaochen;DUAN Chang'en(Department of Cardiovascular Medicine,the First Affiliated Hospital of Xinxiang Medical University,Xinxiang 453100,China)

机构地区：[1]新乡医学院第一附属医院心血管外科,新乡453100 [2]新乡医学院第一附属医院心血管内科,新乡453100

出　　处：《中国免疫学杂志》2022年第20期2455-2460,共6页Chinese Journal of Immunology

基　　金：河南省二〇一八年科技发展计划项目(182102310182)。

摘　　要：目的:探究下调长链非编码RNA核内富集转录物1(lncRNA Neat1)表达对大鼠心肌缺血再灌注损伤的保护作用及其机制。方法:将48只Wistar大鼠随机分为假手术组、模型组、shRNA NC组和sh-Neat1组,采用左前降支结扎法构建心肌缺血再灌注损伤大鼠模型,采用ELISA检测血清心损伤标志物肌红蛋白(Mb)、心肌肌钙蛋白Ⅰ(cTnⅠ)、肌酸激酶同工酶(CK-MB)含量,检测大鼠心率(HR)和左心室射血分数(LVEF),生化试剂盒检测大鼠血清超氧化物歧化酶(SOD)和丙二醛(MDA)含量,HE染色和MASSON染色观察大鼠心肌病理变化,Western blot检测心肌组织中α平滑肌肌动蛋白(α-SMA)、转化生长因子β(TGF-β)、纤维连接蛋白(FN)、核因子NF-κB P65的表达,荧光定量PCR检测心肌组织中lncRNA Neat1、IL-6和TNF-αmRNA的表达,免疫荧光检测NF-κB P65细胞核转位情况。结果:与假手术组比较,模型组大鼠lncRNA Neat1、Mb、cTnⅠ、CK-MB、MDA、IL-6 mRNA、TNF-αmRNA、HR、α-SMA、TGF-β、FN、p-NF-κB P65/NF-κB P65水平升高,LVEF和SOD水平降低(P<0.05);与模型组比较,sh-Neat1组lncRNA Neat1、Mb、cTnI、CK-MB、MDA、IL-6 mRNA、TNF-αmRNA、HR、α-SMA、TGF-β、FN、p-NF-κB P65/NF-κB P65水平降低,LVEF和SOD水平升高(P<0.05)。结论:lncRNA Neat1在心肌缺血再灌注损伤中高表达,干扰lncRNA Neat1后可保护大鼠心肌缺血再灌注损伤,改善心肌纤维化,可能与减少NF-κB P65细胞核转位有关。Objective:To explore the protective effect of down-regulating long non-coding RNA nuclear enriched abundant transcript 1(lncRNA Neat1)against myocardial ischemia-reperfusion injury in rats and its mechanism.Methods:Forty-eight Wistar rats were randomly divided into sham operation group,model group,shRNA NC group and sh-Neat1 group.Myocardial ischemiareperfusion injury rat models were constructed by left anterior descending ligation.The contents of serum myocardial injury markers myoglobin(Mb),cardiac troponinⅠ(cTnⅠ),creatine kinase MB(CK-MB)were detected by ELISA.The heart rate(HR)and left ventricular ejection fraction(LVEF)were detected.The contents of serum superoxide dismutase(SOD)and malondialdehyde(MDA)were detected by biochemical kits.The pathological changes of myocardium were observed by HE staining and MASSON staining.The expressions ofα-smooth muscle actin(α-SMA),transforming growth factor-β(TGF-β),fibronectin(FN)and nuclear factor NF-κB P65 in myocardial tissues were detected by Western blot.The expressions of serum lncRNA Neat1,IL-6 and tumor necrosis factor(TNF-α)mRNA in myocardial tissues were detected by fluorescence quantitative PCR.The nuclear translocation of NF-κB P65 was detected by immunofluorescence.Results:Compared with sham operation group,levels of lncRNA Neat1,Mb,cTnⅠ,CK-MB,MDA,IL-6 mRNA,TNF-αmRNA,HR,α-SMA,TGF-β,FN and p-NF-κB P65/NF-κB P65 were increased,while levels of LVEF and SOD were decreased in model group(P<0.05).Compared with model group,levels of lncRNA Neat1,Mb,cTnⅠ,CK-MB,MDA,IL-6 mRNA,TNF-αmRNA,HR,α-SMA,TGF-β,FN and p-NF-κB P65/NF-κB P65 were decreased,while levels of LVEF and SOD were increased in sh-Neat1 group(P<0.05).Conclusion:lncRNA Neat1 is highly expressed in myocardial ischemia-reperfusion injury.Interfering lncRNA Neat1 can protect myocardial ischemia-reperfusion injury and improve myocardial fibrosis in rats,which may be related to reducing nuclear translocation of NF-κB P65.

关 键 词：lncRNA Neat1 心肌缺血再灌注 NF-κB P65 心室重塑 

分 类 号：R363[医药卫生—病理学]