摘要
目的探究长链非编码RNA HOX转录本反义RNA(lnRNA HOTAIR)靶向结节性硬化症复合体1(TSC1)对糖尿病肾病(DN)足细胞损伤的调控作用。方法将小鼠足细胞MPC5分为6组:对照组、高糖组(HG组)、HG+si-NC组、HG+si-HOTAIR组、HG+si-HOTAIR+sh-NC组、HG+si-HOTAIR+sh-TSC1组。qPCR检测lnRNA HOTAIR及TSC1 mRNA表达水平,细胞计数试剂盒-8(CCK-8)检测细胞活力,流式细胞术检测细胞凋亡,ELISA检测炎症因子[IL-6、肿瘤坏死因子-α(TNF-α)]水平,利用各试剂盒检测氧化应激指标[活性氧(ROS)、丙二醛(MDA)、超氧化物歧化酶(SOD)]水平。结果与对照组比较,HG组lnRNA HOTAIR表达水平、细胞凋亡率,IL-6、TNF-α、ROS及MDA水平明显升高(P<0.05),TSC1 mRNA表达水平、细胞活力、SOD水平明显降低(P<0.05);与HG+si-NC组比较,HG+si-HOTAIR组lnRNA HOTAIR表达水平、MPC5细胞凋亡率,IL-6、TNF-α、ROS及MDA水平明显降低(P<0.05),TSC1 mRNA表达水平、细胞活力、SOD水平明显升高(P<0.05);与HG+si-HOTAIR+sh-NC组比较,HG+si-HOTAIR+sh-TSC1组细胞凋亡率,IL-6、TNF-α、ROS及MDA水平明显升高(P<0.05),TSC1 mRNA水平、细胞活力、SOD水平明显降低(P<0.05)。结论沉默lnRNA HOTAIR可靶向TSC1调控DN足细胞活力、细胞凋亡率、炎症水平及氧化应激水平,进而减轻DN足细胞损伤。
Objective To explore the regulatory effect of long non-coding RNA HOX transcript antisense RNA(lnRNA HOTAIR)targeting tuberous sclerosis complex 1(TSC1)on podocyte injury in diabetic nephropathy(DN).Methods Mice podocyte MPC5 were divided into 6 groups:the control group,the high glucose group(the HG group),the HG+si-NC group,the HG+si-HOTAIR group,the HG+si-HOTAIR+sh-NC group,and the HG+si-HOTAIR+sh-TSC1 group.qPCR was used to detect the levels of lnRNA HOTAIR and TSC1 mRNA.Cell viability was detected by cell counting kit-8(CCK-8),and apoptosis was detected by flow cytometry.ELISA was used to detect the levels of inflammatory factors[IL-6,tumor necrosis factor-α(TNF-α)]and the levels of oxidative stress indicators[reactive oxygen species(ROS),malondialdehyde(MDA),superoxide dismutase(SOD)]were detected by corresponding reagent kit.Results Compared with the control group,the levels of lnRNA HOTAIR,apoptosis rate,IL-6,TNF-α,ROS and MDA in the HG group were significantly increased(P<0.05),while the levels of TSC1 mRNA,cell viability and SOD were significantly decreased(P<0.05).Compared with the HG+si-NC group,the levels of lnRNA HOTAIR,apoptosis rate,IL-6,TNF-α,ROS and MDA in the HG+si-HOTAIR group significantly decreased(P<0.05),the levels of TSC1 mRNA,cell viability and SOD were significant increased(P<0.05).Compared with the HG+si-HOTAIR+sh-NC group,the levels of apoptosis rate,IL-6,TNF-α,ROS and MDA in the HG+si-HOTAIR+sh-TSC1 group were significantly increased(P<0.05),the levels of TSC1 mRNA,cell viability and SOD were significantly decreased(P<0.05).Conclusion Silencing lnRNA HOTAIR can target TSC1 to regulate the activity,apoptosis rate,inflammatory level and oxidative stress levels of podocyte injury in DN,thereby alleviating podocyte damage in DN.
作者
牛秋霞
刘伟丽
李学慧
张茜
王双利
NIU Qiuxia;LIU Weili;LI Xuehui;ZHANG Qian;WANG Shuangli(Department of Gastroenterology,Zhengzhou 460 Hospital,Zhengzhou,Henan 450000,China)
出处
《重庆医学》
2025年第8期1787-1792,共6页
Chongqing Medical Journal
