LncRNA CRNDE调节miR⁃338⁃3p/KLF6轴对LPS诱导的肺泡上皮细胞损伤的影响  

Effect of LncRNA CRNDE on LPS⁃induced alveolar epithelial cell injury by regulating the miR⁃338⁃3p/KLF6 axis

作　　者：岳利英 林雪容 张盟盟 YUE Li-ying;LIN Xue-rong;ZHANG Meng-meng(Department of Blood Transfusion,Zhangjiakou First Hospital,Zhangjiakou 075000,China;Emergency Department,The First Affiliated Hospital of Hebei North University,Zhangjiakou,Hebei Province 075000,China)

机构地区：[1]张家口市第一医院输血科,河北张家口075000 [2]河北北方学院附属第一医院急诊科,河北张家口075000

出　　处：《解剖学研究》2024年第3期228-235,共8页Anatomy Research

基　　金：河北省医学科学研究课题计划(20211689)。

摘　　要：目的探讨LncRNA结直肠肿瘤差异表达基因(CRNDE)调节miR⁃338⁃3p/KLF6轴对LPS诱导的肺泡上皮细胞损伤的影响。方法将人肺泡上皮细胞A549分为CK组、LPS组、LPS+si⁃NC组、LPS+si⁃CRNDE组、LPS+si⁃CRNDE+inhibitor NC组、LPS+si⁃CRNDE+miR⁃338⁃3p inhibitor组、LPS+si⁃CRNDE+oe⁃NC组、LPS+si⁃CRNDE+oe⁃KLF6组;qRT⁃PCR检测各组细胞CRNDE、miR⁃338⁃3p和KLF6表达水平;MTT法检测细胞增殖;流式细胞术检测细胞凋亡率;ELISA试剂盒检测TNF⁃α、IL⁃10和IL⁃1β水平;商品化试剂盒分析MDA、GSH⁃Px、SOD水平;Western blot检测细胞中KLF6、Cleaved⁃caspase⁃3、Bax、Bcl⁃2蛋白表达量。双荧光素酶报告基因实验验证miR⁃338⁃3p与RNDE和KLF6的关系。结果与CK组相比,LPS组和LPS+si⁃NC组A549细胞CRNDE和KLF6 mRNA表达、TNF⁃α、IL⁃1β水平、MDA含量、细胞凋亡率、caspase⁃3、Bax、KLF6蛋白表达显著升高,miR⁃338⁃3p表达、细胞活力、IL⁃10水平、SOD和GSH⁃Px活性、Bcl⁃2蛋白表达显著降低(P<0.05);与LPS+si⁃NC组相比,LPS+si⁃CRNDE组A549细胞CRNDE和KLF6 mRNA表达、TNF⁃α、IL⁃1β水平、MDA含量、细胞凋亡率、caspase⁃3、Bax、KLF6蛋白表达显著降低,miR⁃150⁃5p表达、细胞活力、IL⁃10水平、SOD和GSH⁃Px活性、Bcl⁃2蛋白表达显著升高(P<0.05);干扰miR⁃338⁃3p表达或过表达KLF6均可降低下调CRNDE表达改善LPS诱导A549细胞损伤的作用(P<0.05)。结论下调CRNDE可调控miR⁃338⁃3p/KLF6轴减轻LPS诱导的A549细胞损伤。Objective To investigate the effect of LncRNA CRNDE on LPS⁃induced alveolar epithelial cell injury by regulating the miR⁃338⁃3p/KLF6 axis.Methods Human alveolar epithelial cells A549 were separated into CK group,LPS group,LPS+si⁃NC group,LPS+si⁃CRNDE group,LPS+si⁃CRNDE+inhibitor NC group,LPS+si⁃CRNDE+miR⁃338⁃3p inhibitor group,LPS+si⁃CRNDE+oe⁃NC group,and LPS+si⁃CRNDE+oe⁃KLF6 group;qRT⁃PCR was applied to detect the expression levels of CRNDE,miR⁃338⁃3p,and KLF6 of cells in each group;MTT method was applied to detect cell proliferation;flow cytometry was applied to detect cell apoptosis rate;ELISA kit was applied to detect TNF⁃α,IL⁃10,and IL⁃1βlevels;commercialized reagent kit was applied to analyze MDA,GSH⁃Px,and SOD levels;Western blot was applied to detect the expression levels of KLF6,cleaved⁃caspase⁃3,Bax,and Bcl⁃2 proteins in cells.Dual luciferase reporter gene experiment was applied to verify the relationship between miR⁃338⁃3p and RNDE and KLF6.Results Compared with the CK group,the expression of CRNDE and KLF6 mRNA,the levels of TNF⁃α,IL⁃1β,content of MDA,apoptosis rate,the expression of caspase⁃3,Bax,and KLF6 proteins in A549 cells in the LPS group and LPS+si⁃NC group were obviously increased,the expression of miR⁃338⁃3p,cell viability,the level of IL⁃10,activities of SOD and GSH⁃Px,and expression of Bcl⁃2 protein were obviously reduced(P<0.05);compared with the LPS+si⁃NC group,the expression of CRNDE and KLF6 mRNA,the levels of TNF⁃α,IL⁃1β,content of MDA,apoptosis rate,the expression of caspase⁃3,Bax,and KLF6 proteins in A549 cells in the LPS+si⁃CRNDE group were obviously reduced,the expression of miR⁃338⁃3p,cell viability,the level of IL⁃10,activities of SOD and GSH⁃Px,and expression of Bcl⁃2 protein were obviously increased(P<0.05);interference with miR⁃338⁃3p expression or overexpression of KLF6 was able to reduce the down⁃regulation of CRNDE expression and improve LPS⁃induced A549 cell damage(P<0.05

关 键 词：急性肺损伤 长链非编码RNA 结直肠肿瘤差异表达基因 miR⁃338⁃3p/KLF6轴 脂多糖 肺泡上皮细胞 

分 类 号：R563.8[医药卫生—呼吸系统]