LncRNA SNHG1在同型半胱氨酸致足细胞焦亡中的作用  

Effect of LncRNA SNHG1 on homocysteine⁃induced podocytespyrophosis

作　　者：张正皓 马芳 张晴 夏童童 刘虹麟 白志刚 卢冠军[2,5] 张竞文 彭红建[2,6] 姜怡邓[2] 马胜超 ZHANG Zhenghao;MA Fang;ZHANG Qing;XIA Tongtong;LIU Honglin;BAI Zhigang;LU Guanjun;ZHANG Jingwen;PENG Hongjian;JIANG Yideng;MA Shengchao(School of Basic Medical Sciences,Ningxia Medical University,Yinchuan 750004,China;NHC Key Laboratory of Metabolic Cardiovascular Diseases Research,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学基础医学院,银川750004 [2]国家卫生健康委员会代谢性心血管疾病研究重点实验室,银川750004 [3]宁夏医科大学检验学院,银川750004 [4]宁夏回族自治区人民医院,银川750004 [5]宁夏医科大学总医院,银川750004 [6]中南大学化学化工学院,长沙410083

出　　处：《实用医学杂志》2024年第4期476-482,共7页The Journal of Practical Medicine

基　　金：国家自然科学基金面上项目(编号:82270492);国家自然科学基金地区项目(编号:82060139);国家自然科学基金青年项目(编号:81900273);宁夏医科大学校级重点项目(编号:XZ2022004);宁夏回族自治区重点研发计划项目(编号:2020BFH02001);宁夏回族自治区重点研发一般项目(编号:2019BEG03006);中国医学科学院中央级公益性科研院所基本科研业务费项目(编号:2019PT330002)。

摘　　要：目的探讨lncRNA SNHG1在同型半胱氨酸诱导足细胞焦亡中的作用。方法选取Cbs^(+/-)小鼠随机分成2组,设置正常饮食组(ND)和高蛋氨酸饮食组(HMD),Western blot检测Caspase-1、Cleaved Caspase-1和NLRP3的蛋白表达水平。体外培养小鼠肾小球足细胞,设置对照组(Control,0μmol/L Hcy)和同型半胱氨酸干预组(Hcy,80μmol/L Hcy);足细胞转染慢病毒后设置lncSNHG1过表达阴性对照组(OE-NC)和lncSNHG1过表达组(OE-SNHG1);lncSNHG1过表达阴性对照+同型半胱氨酸干预组(OE-NC+Hcy)和lncSNHG1过表达+同型半胱氨酸干预组(OE-SNHG1+Hcy),干预细胞48 h后,实时荧光定量PCR检测Hcy干预的足细胞中lncSNHG1表达水平;Western blot和免疫荧光技术检测足细胞中焦亡相关蛋白Caspase-1、Cleaved Caspase-1、NLRP3及GSDMD和GSDMD-N表达水平;ELISA检测足细胞中焦亡介导的炎症相关因子IL-1β和IL-18含量。结果(1)动物实验中:与正常饮食组(ND)相比,高蛋氨酸饮食组(HMD)中焦亡相关蛋白Caspase-1、Cleaved Caspase-1和NLRP3及GSDMD、GSDMD-N表达水平增高;(2)细胞实验中:与对照组(Control)相比,同型半胱氨酸干预组(Hcy)中焦亡相关蛋白Caspase-1、Cleaved Caspase-1和NLRP3及GSDMD、GSDMD-N表达水平增高,焦亡介导炎症因子IL-1β和IL-18含量增高;(3)细胞实验中:与对照组(Control)相比,同型半胱氨酸干预组(Hcy)中lncSNHG1表达水平增高;足细胞转染lncSNHG1慢病毒后,与Control组及OE-NC组相比,OE-SNHG1组lncSNHG1表达水平增高;(4)细胞实验中:与OE-NC+Hcy组相比,OE-SNHG1+Hcy组中焦亡相关蛋白Caspase-1、Cleaved Caspase-1和NLRP3及GSDMD、GSDMD-N表达水平增高,焦亡介导炎症因子IL-1β和IL-18含量增高。结论lncRNA SNHG1在Hcy诱导的足细胞焦亡过程中起到促进作用。Objective To investigate the role of lncRNA SNHG1 in homocysteine-induced pyroptosis of podocyte.Methods Cbs^(+/-)mice were randomly divided into two groups:a normal diet group(ND)and a high me⁃thionine diet group(HMD).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocytes were cultured in vitro,and then assigned into a control group(Control,0μmol/L Hcy)and a homocysteine intervention group(Hcy,80μmol/L Hcy).Western blotting was used to detect the protein expression levels of Caspase-1,Cleaved Caspase-1,and NLRP3.Mouse renal glomerular podocyion group(OE-NC+Hcy)and the lncSNHG1 overexpression+homocysteine intervention group(OE-SNHG1+Hcy)were also established.After 48 hours of intervention,Real-time fluorescence quantita⁃tive PCR was used to detect the expression of lncSNHG1 in podocytes after Hcy intervention.Western blot was used to detect the expressions of Caspase-1,Cleaved Caspase-3 and NLRP3.Immunofluorescence was used to de⁃tect the expression levels of GSDMD and GSDMD-N.ELISA was used to detect the contents of IL-1βand IL-18.Results(1)In the animal experiments,the expression levels of pyroptosis-related proteins Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the HMD group compared with the ND group.(2)In the cellular experiments,the expression levels of Caspase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were all increased in the Hcy group compared with the Control group,and the contents of pyroptosismediated inflammatory factors IL-1βand IL-18 were increased as well.(3)In the cellular experiments,the expres⁃sion of lncSNHG1 was increased in the Hcy group compared with the control group.After transduction with lnc⁃SNHG1 lentivirus,the expression of lncSNHG1 was increased in the OE-SNHG1 group,compared with the control group and the OE-NC group.(4)In the cellular experiments,the expressions of pyroptosis-related proteins Cas⁃pase-1,Cleaved Caspase-1,NLRP3,GSDMD,and GSDMD-N were inc

关 键 词：同型半胱氨酸 足细胞 核仁小RNA宿主基因1 焦亡 

分 类 号：R34[医药卫生—基础医学]