VD3调控ROS介导的TXNIP/NLRP3通路抑制高糖诱导肾系膜细胞纤维化的机制研究  

VD3 Regulates ROS Mediated TXNIP/NLRP3 Pathway to Inhibit Hyperglycemia-Induced Renal Mesangial Cell Fibrosis

作　　者：刘刚[1] 孟庆悦 张春江[1] 杨晓萍[1] LIU Gang(The First Affiliated Hospital of Shihezi University,Xinjiang Shihezi 832008,China)

机构地区：[1]石河子大学第一附属医院肾病内科,新疆石河子832008 [2]石河子大学医学院,新疆石河子832008

出　　处：《河北医学》2024年第5期762-768,共7页Hebei Medicine

基　　金：石河子大学自主资助支持立项校级科研项目,(编号:ZZZC202192)。

摘　　要：目的:探究活性维生素D3(VD3)对高糖诱导下肾系膜细胞纤维化特征的影响,并探讨相关作用机制。方法:培养大鼠肾小球系膜细胞系HBZY-1,将其分为正常葡萄糖培养(NG)组、正常葡萄糖联合VD3培养(NG+VD3)组、高葡萄糖培养(HG)组、高葡萄糖联合VD3培养(HG+VD3)组、高葡萄糖联合N-乙酰半胱氨酸培养(HG+NAC)组。CCK-8法检测各组细胞的增殖活性,流式细胞术检测各组细胞的凋亡率,DCFH-DA法检测各组细胞中活性氧(ROS)含量,酶联免疫吸附法(ELISA)检测各组细胞上清液中白细胞介素-1β(IL-1β)、IL-18、肿瘤坏死因子-α(TNF-α)水平,实时荧光定量聚合酶反应(RT-qPCR)检测各组细胞中纤维化标志物转化生长因子-β1(TGF-β1)、纤维连接蛋白(FN)、细胞间粘附分子-1(ICAM-1)及硫氧还蛋白互作蛋白(TXNIP)、NOD样受体热蛋白结构域相关蛋白3(NLRP3)的mRNA表达,蛋白质免疫印记(Western blot)检测各组细胞中TGF-β1、FN、ICAM-1及TXNIP、NLRP3的蛋白表达。结果:与NG组比较,HG组增殖活性显著升高(P<0.05),细胞凋亡率显著减少(P<0.05),ROS含量显著上升(P<0.05),细胞上清液中IL-1β、IL-18、TNF-α水平显著升高(P<0.05),细胞中TGF-β1、FN、ICAM-1、TXNIP、NLRP3的mRNA和蛋白相对表达水平显著上调(P<0.05);与HG组比较,HG+VD3组和HG+NAC组增殖活性显著降低(P<0.05),细胞凋亡率显著增加(P<0.05),ROS含量显著下降(P<0.05),细胞上清液中IL-1β、IL-18、TNF-α水平显著降低(P<0.05),同时,细胞中TGF-β1、FN、ICAM-1及TXNIP、NLRP3的mRNA和蛋白相对表达水平显著下调(P<0.05)。结论:VD3能够减轻高糖诱导的肾系膜细胞过度增殖及纤维化,该机制可能与调控ROS/TXNIP/NLRP3通路有关。Objective:To investigate the effect of active vitamin D3(VD3)on the characteristics of renal mesangial cell fibrosis induced by high glucose,and to explore the relevant mechanism.Methods:Rat mesangial cell line HBZY-1 was cultured and divided into normal glucose culture(NG)group,normal glucose combined with VD3 culture(NG+VD3)group,high glucose culture(HG)group,high glucose combined with VD3 culture(HG+VD3)group,high glucose combined with n-acetylcysteine culture(HG+NAC)group.The proliferation activity of cells in each group was detected by CCK-8 method,the apoptosis rate of cells in each group was detected by flow cytometry,the content of reactive oxygen species(ROS)in cells in each group was detected by DCFH-DA method,the levels of interleukin-1β(IL-1β),IL-18 and tumor necrosis factor-α(TNF-α)in cell supernatant were detected by enzyme-linked immunosorbent assay(ELISA),the mRNA expression and the protein expression of transforming growth factor-β1(TGF-β1),fibronectin(FN),intercellular adhesion molecule-1(ICAM-1),thioredoxin interacting protein(TXNIP),NOD-like receptor heat protein domain associated protein 3(NLRP3)in cells in each group were detected by real-time fluorescent quantitative polymerase reaction(RT-qPCR)and Western blot.Results:Compared with NG group,the proliferative activity in HG group was significantly increased(P<0.05),the apoptosis rate was significantly decreased(P<0.05),ROS content was significantly increased(P<0.05),the levels of IL-1β,IL-18 and TNF-αin cell supernatant were significantly increased(P<0.05),and the mRNA and protein relative expression levels of TGF-β1,FN,ICAM-1,TXNIP and NLRP3 were significantly up-regulated(P<0.05).Compared with HG group,the proliferative activity in HG+VD3 and HG+NAC groups was significantly decreased(P<0.05),the apoptosis rate was significantly increased(P<0.05),the ROS content was significantly decreased(P<0.05),the levels of IL-1β,IL-18 and TNF-αin cell supernatant were significantly decreased(P<0.05),and the mRNA and protein relative express

关 键 词：糖尿病肾病 肾系膜细胞 活性维生素D3 增殖 纤维化 

分 类 号：R587.2[医药卫生—内分泌] R692[医药卫生—内科学]