FOXM1通过靶向上调PHB2的转录改善线粒体功能障碍保护脂多糖诱导的肾小管上皮细胞损伤  

FOXM1 Ameliorates Mitochondrial Dysfunction by Targeting Upregulation of PHB2 Transcription to Protect Against Lipopolysaccharide-Induced Renal Tubular Epithelial Cell Injury

作　　者：陈娟[1] 陈拉斯 陈丽 唐文庄 张琼果 王善志 CHEN Juan;CHEN Lasi;CHEN Li(The First Affiliated Hospital of Hainan Medical College, Hainan Haikou 570000, China)

机构地区：[1]海南医学院第一附属医院,海南海口570000

出　　处：《河北医学》2022年第3期368-374,共7页Hebei Medicine

基　　金：国家自然科学基金项目,(编号:81560124);海南省卫生健康行业科研项目,(编号:20A200165)。

摘　　要：目的:探究FOXM1在脂多糖(LPS)诱导的肾小管上皮细胞损伤中的作用及其可能的作用机制。方法:体外培养人肾小管上皮细胞HK-2,qRT-PCR和Western blot检测细胞FOXM1和PHB2表达;CCK-8检测细胞活力;流式细胞术检测细胞凋亡;JC-1染色检测线粒体膜电位;试剂盒检测细胞ATP含量;荧光素酶报告实验和ChIP-PCR实验检测FOXM1对PHB2启动子调控作用。结果:与对照组相比,LPS组细胞中FOXM1和PHB2表达、细胞活力、红/绿荧光强度比值和ATP含量明显降低(P<0.05),细胞凋亡率明显升高(P<0.05);与LPS组比较,LPS+oe-NC组细胞中FOXM1和PHB2表达、细胞活力、红/绿荧光强度比值、ATP含量和细胞凋亡率差异无统计学意义(P>0.05);与LPS+oe-NC组相比,LPS+oe-FOXM1组细胞中FOXM1和PHB2表达、细胞活力、红/绿荧光强度比值和ATP含量明显升高(P<0.05),细胞凋亡率明显降低(P<0.05)。FOXM1靶向结合PHB2启动子区域。与LPS+oe-NC+sh-NC组相比,LPS+oe-FOXM1+sh-NC组细胞中FOXM1和PHB2蛋白表达、细胞活力、红/绿荧光强度比值和ATP含量明显升高(P<0.05),细胞凋亡率明显降低(P<0.05);与LPS+oe-NC+sh-NC组相比,LPS+oe-NC+sh-PHB2组细胞中PHB2蛋白表达、细胞活力、红/绿荧光强度比值和ATP含量明显降低(P<0.05),细胞凋亡率明显升高(P<0.05),FOXM1蛋白表达差异无统计学意义(P>0.05);sh-PHB2可部分逆转oe-FOXM1对LPS处理的HK-2细胞的作用。结论:FOXM1通过靶向结合PHB2启动子区域促进PHB2表达,改善线粒体功能障碍,从而抑制LPS诱导的肾小管上皮细胞损伤。Objective:To explore the role of FOXM1 in lipopolysaccharide(LPS)-induced renal tubular epithelial cell injury and its possible mechanism.Methods:Human renal tubular epithelial cells HK-2 were cultured in vitro.QRT-PCR and Western blot were used to detect the expression of FOXM1 and PHB2;CCK-8 was used to detect cell viability;flow cytometry was used to detect cell apoptosis;JC-1 staining was used to detect the mitochondrial membrane potential;the reagent kit was used to detect cellular ATP content;the luciferase reporter experiment and ChIP-PCR experiment were used to detect the regulatory effect of FOXM1 on the PHB2 promoter.Results:Compared with the control group,the expression of FOXM1 and PHB2,cell viability,red/green fluorescence intensity ratio and ATP content in the cells of the LPS group were significantly reduced(P<0.05),and the apoptosis rate was significantly increased(P<0.05);compared with the LPS group,there was no significant difference in FOXM1 and PHB2 expression,cell viability,red/green fluorescence intensity ratio,ATP content and apoptosis rate in the cells of the LPS+oe-NC group(P>0.05);compared with the LPS+oe-NC group,the expression of FOXM1 and PHB2,cell viability,red/green fluorescence intensity ratio and ATP content in the cells of the LPS+oe-FOXM1 group were significantly increased(P<0.05),and the apoptosis rate was significantly reduced(P<0.05).FOXM1 targeted the PHB2 promoter region.Compared with the LPS+oe-NC+sh-NC group,the FOXM1 and PHB2 protein expression,cell viability,red/green fluorescence intensity ratio and ATP content in the cells of the LPS+oe-FOXM1+sh-NC group were significantly increased(P<0.05),the cell apoptosis rate was significantly reduced(P<0.05);compared with the LPS+oe-NC+sh-NC group,the expression of PHB2 protein,cell viability,red/green fluorescence intensity ratio and ATP content in the cells of the LPS+oe-NC+sh-PHB2 group were significantly reduced(P<0.05),the apoptosis rate was significantly increased(P<0.05),and the difference in FOXM1 protein expression was

关 键 词：肾小管上皮细胞 脂多糖 Forkhead box M1 prohibitin 2 线粒体 

分 类 号：Q78[生物学—分子生物学]