长链非编码RNA MEG3吸附miR-29a通过NF-κB/NLRP3通路参与糖尿病肾病大鼠足细胞焦亡  

Long Chain Noncoding RNA MEG 3 Adsorbs miR-29 a and Participates in Podocyte Pyroptosis in Diabetes Nephropathy Rats through NF-κB/NLRP3 Pathway

作　　者：袁祖君 王倩倩[1] 张全军 林昌伟 YUAN Zujun;WANG Qianqian;ZHANG Quanjun;LIN Changwei(Department of Nephrology,Beijing Anzhen Nanchong Hospital,Capital Medical University&Nanchong Central Hospital,Nanchong 637000,China)

机构地区：[1]首都医科大学附属北京安贞医院南充医院·南充市中心医院肾内科,南充637000

出　　处：《福建医科大学学报》2024年第6期357-366,共10页Journal of Fujian Medical University

基　　金：南充市科学技术局项目(22SXQT0135)。

摘　　要：目的探讨长链非编码RNA母系表达基因3(lncRNA MEG 3)吸附miR-29 a对核转录因子-κB/NOD样受体热蛋白结构域相关蛋白3(NF-κB/NLRP3)通路及糖尿病肾病(DN)大鼠足细胞焦亡的影响。方法96只大鼠随机分为空白对照(Blank)组、DN组、敲低对照(sh-NC)组、敲低lncRNA MEG 3表达(sh-MEG 3)组,过表达对照(miR-NC)组、过表达miR-29 a(miR-29 a)组、敲低lncRNA MEG 3表达+敲低对照(sh-MEG3+anti-miR-NC)组、敲低lncRNA MEG 3表达+敲低miR-29 a表达(sh-MEG 3+anti-miR-29 a)组,每组各12只。采用高糖、高脂饮食联合注射链脲佐菌素(STZ)制备DN大鼠模型。sh-NC组、sh-MEG 3组、miR-NC组、miR-29 a组、sh-MEG 3+anti-miR-NC组、sh-MEG 3+anti-miR-29 a组大鼠通过尾静脉注射100μL相应的慢病毒(均为1012 TU/mL)。注射4周后,生化分析仪检测空腹血糖(FBG)、血清总胆固醇(TC)、甘油三酯(TG)、血肌酐(Scr)、尿素氮(BUN)和尿微量白蛋白(UMA)水平;PAS染色观察肾小球损伤;透射电镜观察足细胞损伤;免疫荧光染色、RT-qPCR、Western-blot检测肾组织突触足蛋白(Synaptopodin)、肾病蛋白(Nephrin)、足突蛋白(Podocin)、lncRNA MEG 3、miR-29 a、NF-κB p 65、NLRP 3、含半胱氨酸的天冬氨酸蛋白水解酶-1(Caspase-1)、白细胞介素(IL)-18、IL-1β、Gasdermin家族蛋白D(GSDMD)mRNA和蛋白水平。结果与Blank组比较,DN组大鼠肾组织lncRNA MEG 3水平升高,miR-29 a水平降低,FBG、血清TC、TG、Scr、BUN和UMA水平升高,肾小球肥大,可见足细胞足突丢失和融合,基底膜明显增厚,肾组织中Synaptopodin、Nephrin、Podocin水平降低,NF-κB p 65、NLRP 3、Caspase-1、IL-18、IL-1β、GSDMD mRNA和蛋白水平升高(均P<0.05);敲低lncRNA MEG 3表达或过表达miR-29 a后,大鼠肾组织lncRNA MEG 3水平降低,miR-29 a水平升高,FBG、血清TC、TG、Scr、BUN和UMA水平降低,肾小球结构基本正常,足细胞仅有少量足突丢失和融合,基底膜轻微增厚,肾组织中Synaptopodin、Nephrin、Podocin水平升Objective To explore the effects of long chain noncoding RNA maternally expressed gene 3(lncRNA MEG 3)on nuclear transcription factor-κB/NOD like receptor thermal protein domain associated protein 3(NF-κB/NLRP3)pathway and podocyte pyroptosis in diabetes nephropathy(DN)rats.Methods Ninety-six rats were randomly divided into Blank group,DN group,knockdown control group(sh-NC),knockdown lncRNA MEG 3 expression group(sh-MEG 3),overexpression control group(miR-NC),overexpression miR-29 a group(miR-29 a),knockdown lncRNA MEG 3 expression+knockdown control group(sh-MEG 3+anti-miR-NC),knockdown lncRNA MEG 3 expression+knockdown miR-29 a expression group(sh-MEG 3+anti-miR-29 a),with 12 rats in each group.The combination of high sugar and high-fat diet with injection of streptozotocin(STZ)was used to prepare diabetic rat model.Rats in sh-NC group,sh-MEG 3 group,miR-NC group,miR-29 a group,sh-MEG 3+anti-miR-NC group,and sh-MEG 3+anti-miR-29 a group were injected with 100μL of corresponding lentivirus via tail vein(all 1012 TU/mL).After one month of injection,the biochemical analyzer was used to detect fasting blood glucose(FBG),serum total cholesterol(TC),triglycerides(TG),serum creatinine(Scr),urea nitrogen(BUN),and urinary microalbumin(UMA)levels;PAS staining was used to observe glomerular injury;transmission electron microscopy was used to observe podocyte injury;immunofluorescence staining,RT-qPCR,Western-blot analysis were used to detect Synaptopodin,Nephrin,Podocin proteins,lncRNA MEG 3,miR-29 a,NF-κB p 65,NLRP 3,Caspase-1,IL-18,IL-1β,GSDMD mRNA and protein levels in renal tissue.Results Compared with Blank group,lncRNA MEG 3 level in renal tissue of DN group rats was increased,miR-29 a level was decreased,FBG,serum TC,TG,Scr,BUN,and UMA levels were increased,Glomerular hypertrophy,visible loss and fusion of podocyte processes,and significant thickening of the basement membrane,Synaptopodin,Nephrin,and Podocin levels in renal tissue were decreased,while NF-κB p 65,NLRP 3,Caspase-1,IL-18,IL-1β,GSDMD mRNA and pr

关 键 词：lncRNA MEG 3 miR-29 a NF-κB/NLRP3通路 糖尿病肾病 细胞焦亡 

分 类 号：R73[医药卫生—肿瘤]