实用医学杂志 ›› 2023, Vol. 39 ›› Issue (1): 12-20.doi: 10.3969/j.issn.1006⁃5725.2023.01.003

• 基础研究 • 上一篇    下一篇

LncRNA H19调控肾小球足细胞焦亡中作用

汪乐新1,3,4 刘超1,3,4 马天龙1,2 杨安宁1,2,3 熊建团1,2,3 吴凯1,2,3 焦运1 白志刚1,2,3 姜怡邓1,2,3 马胜超1,2,3 张旭4 卢冠军1,4   

  1. 宁夏医科大学1 临床医学院,2 基础医学院(银川 750004);3 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);4 宁夏医科大学总医院泌尿外科(银川 750004); 5 中国人民解放军总医院泌尿外科(北京 100000)

  • 出版日期:2023-01-10 发布日期:2023-01-10
  • 通讯作者: 卢冠军 E⁃mail:sssluguanjun566@163.com
  • 基金资助:
    国家自然科学项目(编号:82060139,81900273);宁夏回族自治区重点研发计划项目(编号:2020BFH02003,2021BEG02033,2021BEG02028,2020BEG03005,2020BFH02001);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)

Role of LncRNA H19 in pyrophosis of glomerular podocyte induced by high glucose

WANG Lexin* ,LIU Chao, MA Tianlong,YANG Anning,XIONG Jiantuan,WU Kai,JIAO Yun,BAI Zhigang,JIANG Yideng,MA Shengchao, ZHANG Xu,LU Guanjun   

  1. Clinical Medical College of Ningxia Medical University,Yinchuan 750004,China;*Key Laboratory of Metabolic Cardiovascular Disease Research of National Health Commission,Yinchuan 750004,China *Department of Urology,General Hospital of Ningxia Medical University,Yinchuan 750004,China 

  • Online:2023-01-10 Published:2023-01-10
  • Contact: LU Guanjun E⁃mail:luguanjun566@163.com

摘要:

目的 探讨 lncRNA H19 在高糖调控肾小球足细胞焦亡中的作用。方法 qRT⁃PCR 方法检 测高糖中 lncRNA H19 本底表达;Western blot 检测高糖中肾足细胞结构性相关蛋白 Nephrin、Podocin 表达; Western blot 及免疫荧光方法检测细胞焦亡相关蛋白 NLRP3、Caspase⁃1、IL⁃1β 表达水平;qRT⁃PCR 检测转 Ad lncRNA H19 si lncRNA H19 lncRNA H19 mRNA 表达,细胞转染分别设置:(1)对照组(不做处 理)、lncRNA H19 过表达阴性对照组、lncRNA H19 过表达组;(2)对照组(不做处理)、lncRNA H19 干扰阴性 对照组和 lncRNA H19 干扰组(si⁃lncRNA H19⁃1、si⁃lncRNA H19⁃2、si⁃lncRNA H19⁃3);Western blot 检测转染 Ad lncRNA H19及si lncRNA H19后NLRP3、Caspase⁃1、IL⁃1β 蛋白表达水平,细胞转染分别设置:(1)对照组 (不做处理)、高糖组、lncRNA H19 过表达对照+高糖组、lncRNA H19 过表达+高糖组;(2)对照组(不做处 理)、高糖组、lncRNA H19 干扰对照+高糖组、lncRNA H19 干扰+高糖组。结果 (1)与对照组(不做处理) 相比,lncRNA H19 在高糖诱导肾足细胞损伤过程中呈低表达(P < 0.05),而 lncRNA H19 低表达可调控细 胞焦亡相关蛋白 NLRP3、Caspase⁃1、IL⁃1β 表达增高(P < 0.05),lncRNA H19 表达与细胞焦亡相关蛋白表达 两者呈负相关;(2)与对照组(不做处理)相比,在高糖诱导肾足细胞损伤过程中,肾足细胞结构蛋白Nephrin Podocin 表达水平降低(P < 0.05);(3)与 lncRNA H19 过表达对照+高糖组相比,过表达 lncRNA H19 后,缓 解高糖诱导肾足细胞焦亡,细胞焦亡相关蛋白 NLRP3、Caspase⁃1、IL⁃1β 呈低表达(P < 0.05);(4)与 lncRNA H19 干扰对照+高糖组相比,干扰 lncRNA H19 后,促进高糖诱导足细胞焦亡,而焦亡相关蛋白 NLRP3 Caspase⁃1、IL⁃1β 呈高表达(P < 0.05)。结论 在高糖诱导足细胞焦亡过程中 lncRNA H19 具有可调控细胞 焦亡相关蛋白表达,为未来治疗糖尿病肾病提供一定的理论依据。

关键词:

LncRNAH19, 糖尿病肾病, 肾足细胞损伤, 细胞焦亡

Abstract:

Objective To investigate the role of lncRNA H19 in pyrophosis of glomerular podocyte induced by high glucose(HG). Methods The background expression of lncRNA H19 in HG was detected by qRT PCR;western blot was used to detect the expression of Nephrin and Podocin;detection of NLRP3,Caspase⁃1 and IL⁃1β expression level was performed by using western blot and immunofluorescence. QRT⁃PCR was used to detect the mRNA expression of lncRNA H19 after transfection of Ad lncRNA H19 and si lncRNA H19. The cell transfec⁃ tion were set as follows:(1)control group(without treatment),negative control group of lncRNA H19 overexpres⁃ sion,lncRNA H19 overexpression group;(2)control group(without treatment),negative control group of lncRNA H19 interference and lncRNA H19 interference group(si lncRNA H19⁃1,si lncRNA H19⁃2,si lncRNA H19⁃3). Detection of NLRP3,Caspase⁃1 and IL⁃1βexpression level after transfection with Ad lncRNA H19 and si lncRNA H19 was performed by western blot. The cell transfection were set as follows:(1)control group(without treatment)high glucose group,lncRNA H19 overexpression control+high glucose group,lncRNA H19 overexpression+high glucose group,(2)control group(without treatment),high glucose group,lncRNA H19 interference control+high glucose group,and lncRNA H19 interference+high glucose group. Results (1)Compared with that in the control group(without treatment),the expression of lncRNA H19 was low in the process of renal podocyte injury induced by high glucose(P < 0.05),and the low expression of lncRNA H19 led to higher expression of pyrophosis⁃related proteins NLRP3,Caspase⁃1,and IL⁃1 β. There was a negative correlation between the expression of lncRNA H19 and the expression of pyrophosis⁃related proteins.(2)Compared with that in the control group(without treatment), the expression level of Nephrin and Podocin in renal podocytes decreased during the process of injury induced by high glucose(P < 0.05).(3)Compared with that in the lncRNA H19 overexpression control+ high glucose group overexpression of lncRNA H19 alleviated high glucose induced nephropodocyte pyrophosis,and the expression of pyrophosis⁃related proteins NLRP3,Caspase⁃1,and IL⁃1 β was low(P < 0.05).(4)Compared with that in the lncRNA H19 interference control+high glucose group,the interference of lncRNA H19 promoted the high glucose⁃ induced podocyte pyrophosis,while the pyrophosis⁃related proteins NLRP3,Caspase⁃1,and IL⁃1 β were highly expressed(P < 0.05). Conclusion In the process of podocyte pyrophosis induced by high glucose,lncRNA H19 can regulate the expression of pyrophosis⁃related proteins,which provides theoretical basis for the treatment of dia⁃ betic nephropathy in the future.

Key words:

LncRNAH19, diabetic nephropathy, renal podocyte damage, cell scorch