LncRNA SNHG1在同型半胱氨酸致足细胞焦亡中的作用OACSTPCD
Effect of LncRNA SNHG1 on homocysteine-induced podocytespyrophosis
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,0 µmol/L Hcy)和同型半胱氨酸干预组(Hcy,80 µmol/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,0 µmol/L Hcy)and a homocysteine intervention group(Hcy,80 µmol/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-1β and 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 pyroptosis-mediated inflammatory factors IL-1β and 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 increased compared with the OE-NC+Hcy group,and the contents of pyroptosis-mediated inflammatory factors IL-1β and IL-18 were increased in the OE-SNHG1+Hcy group.Conclusion These results indicate that lncSNHG1 may play a role in promoting Hcy induced podocytepyroptosis.
张正皓;姜怡邓;马胜超;马芳;张晴;夏童童;刘虹麟;白志刚;卢冠军;张竞文;彭红建
宁夏医科大学基础医学院, 银川 750004||宁夏医科大学检验学院 银川 750004宁夏医科大学检验学院 银川 750004宁夏医科大学检验学院 银川 750004||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宁夏医科大学检验学院 银川 750004||宁夏回族自治区人民医院 银川 750004宁夏医科大学检验学院 银川 750004||宁夏医科大学总医院 银川 750004宁夏医科大学基础医学院, 银川 750004宁夏医科大学检验学院 银川 750004||中南大学化学化工学院 长沙 410083
基础医学
同型半胱氨酸足细胞核仁小RNA宿主基因1焦亡
homocysteinepodocytesmall nucleolar RNA host gene 1pyroptosis
《实用医学杂志》 2024 (004)
476-482 / 7
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)
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