毛蕊花糖苷调节肌醇需要酶1α-硫氧还蛋白相互作用蛋白-Nod样受体蛋白3信号通路减轻脂多糖诱导的肾小球血管内皮细胞损伤  

Acteoside on lipopolysaccharide-induced human glomerular endothelial cell injury by regulating the IRE1α-TXNIP-NLRP3 signaling pathway

作　　者：荆堂堂 李伟伟 王禹婷 孙梦鸽 JING Tangtang;LI Weiwei;WANG Yuting;SUN Mengge(Department of Clinical Laboratory,Shanghai Traditional Chinese Medicine Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 200082,China)

机构地区：[1]上海中医药大学附属市中医医院检验科,上海200082

出　　处：《肾脏病与透析肾移植杂志》2025年第1期38-44,共7页Chinese Journal of Nephrology,Dialysis & Transplantation

基　　金：上海市宝山区2022年度院级科研课题(22-A-5)。

摘　　要：目的:探讨毛蕊花糖苷(Act)调控肌醇需要酶1α(IRE1α)-硫氧还蛋白相互作用蛋白(TXNIP)-Nod样受体蛋白3(NLRP3)信号轴对脂多糖(LPS)诱导的肾小球血管内皮细胞(GEC)损伤的影响。方法:RT-qPCR和Western Blot实验检测转染细胞中IRE1αmRNA和蛋白表达水平;将人肾小球内皮细胞(HRGEC)分为对照组、LPS组、低剂量Act组(Act-L组,50μmol/L)、高剂量Act组(Act-H组,100μmol/L)、Act-H+pcDNA-NC组和Act-H+pcDNA-IRE1α组;试剂盒检测HRGEC中活性氧(ROS)生成量;CCK-8法检测HRGEC存活情况;流式细胞术检测HRGEC凋亡情况;ELISA试验测定HRGEC中白细胞介素(IL)-1β、IL-6、IL-18和肿瘤坏死因子α(TNF-α)水平;Western Blot测定IRE1α、p-IRE1α、TXNIP和NLRP3蛋白表达量。结果:与对照组相比,LPS组HRGEC中ROS生成量、细胞凋亡率、炎性分子IL-1β、IL-6、IL-18和TNF-α水平及p-IRE1α/IRE1α、TXNIP、NLRP3蛋白表达量均显著升高(P<0.05),细胞存活率均显著降低(P<0.05);与LPS组相比,Act-L组、Act-H组、Act-H+pcDNA-NC组HRGEC中ROS生成量、细胞凋亡率、IL-1β、IL-6、IL-18和TNF-α水平及p-IRE1α/IRE1α、TXNIP、NLRP3蛋白表达量均显著降低(P<0.05),细胞存活率均显著升高(P<0.05);与Act-H+pcDNA-NC组相比,Act-H+pcDNA-IRE1α组中ROS生成量、细胞凋亡率、IL-1β、IL-6、IL-18和TNF-α水平及p-IRE1α/IRE1α、TXNIP、NLRP3蛋白表达量均显著升高(P<0.05),细胞存活率均显著降低(P<0.05)。结论:Act可能通过下调IRE1α-TXNIP-NLRP3信号通路中关键蛋白的表达,缓解LPS诱导的GEC损伤,促进细胞增长,抑制炎性分子分泌和细胞凋亡。Objective:To investigate the effect of acteoside(Act)on lipopolysaccharide(LPS)-induced injury to glomerular endothelial cells(GEC)by regulating the inositol-requiring enzyme 1α(IRE1α)-thioredoxin-interacting protein(TXNIP)-NOD-like receptors family pyrin domain containing 3(NLRP3)signaling axis.Methodology:RT qPCR and Western blot experiments were used to detect the expression levels of IRE1αmRNA and protein in transfected cells.The human renal GEC(HRGEC)cells were separated into control(Ctrl)group,LPS group,low-dose Act group(Act-L,50μmol/L),high-dose Act group(Act-H,100μmol/L),Act-H+pcDNA-NC group,and Act-H+pcDNA-IRE1αgroup.The reagent kit was used to detect the production of reactive oxygen species(ROS)in HRGEC cells.CCK-8 method was applied to detect the survival of HRGEC cells.Flow cytometry was applied to detect apoptosis in HRGEC cells.ELISA assay was applied to measure the levels of IL-1β,IL-6,IL-18 and TNF-α.TNF-αin HRGEC cells.Western blot was applied to determine the expression levels of IRE1α,p-IRE1α,TXNIP,and NLRP3 proteins.Results:compared with the Ctrl group,the ROS production,apoptosis rate,the levels of inflammatory molecules IL-1β,IL-6,IL-18 and TNF-α,and the expression levels of p-IRE1α/IRE1α,TXNIP,and NLRP3 proteins in HRGEC cells in the LPS group were obviously higher(P<0.05),and the cell survival rate was obviously lower(P<0.05).And then,compared with the LPS group,the ROS production,apoptosis rate,the levels of IL-1β,IL-6,IL-18 and TNF-α,and the expression levels of p-IRE1α/IRE1α,TXNIP,and NLRP3 proteins in HRGEC cells in the Act-L group,Act-H group,and Act-H+pcDNA-NC group were obviously lower(P<0.05),and the cell survival rate was obviously higher(P<0.05).In the end,compared with the Act-H+pcDNA-NC group,the ROS production,apoptosis rate,the levels of IL-1β,IL-6,IL-18 and TNF-α,and the expression levels of p-IRE1α/IRE1α,TXNIP,and NLRP3 proteins in the Act-H+pcDNA-IRE1αgroup were obviously higher(P<0.05),while the cell survival rate was obviously lower(P<0.05).Conc

关 键 词：毛蕊花糖苷 肌醇需要酶1α-硫氧还蛋白相互作用蛋白-Nod样受体蛋白3信号轴 肾小球血管内皮细胞 

分 类 号：R692[医药卫生—泌尿科学]