卡格列净通过抑制活性氧/NOD样受体热蛋白结构域相关蛋白3信号通路减缓足细胞损伤
Canagliflozin attenuates human podocyte injury through inhibiting reactive oxygen species/NOD-like receptor thermal protein domain associated protein 3 signaling pathway
程艳璐 1陈瑛 1王雅楠 1李思雨 2王利 3王浩 1朱冰冰4
作者信息
- 1. 上海中医药大学附属普陀医院肾内科,上海 200062
- 2. 安徽医科大学上海普陀中心临床学院,上海 200062
- 3. 上海中医药大学附属普陀医院肾内科,上海 200062;安徽医科大学上海普陀中心临床学院,上海 200062
- 4. 上海中医药大学附属普陀医院肾内科,上海 200062;安徽医科大学上海普陀中心临床学院,上海 200062;上海交通大学医学院附属第六人民医院肾内科,上海 200233
- 折叠
摘要
目的 观察卡格列净(canagliflozin,Cana)干预治疗高糖诱导人足细胞(human podocyte,HPC)损伤的疗效,并探讨其相关机制.方法 将HPC设为5组:正常组(normal glucose group,NG组)、甘露醇组(mannitol group,MA组)、高糖组(high glucose,HG组)、Cana低剂量(0.3 μmol/L)组、Cana高剂量(1.0 μmol/L)组.Western印迹法检测膜相关反向鸟苷酸激酶2(membrane-associated guanylate kinase inverted-2,MAGI2)、足细胞相关蛋白 nephrin、钠-葡萄糖转运蛋白 2(sodium-glucose transporter 2,SGLT2)及 NOD 样受体热蛋白结构域相关蛋白 3(NOD-like receptor thermal protein domain associated protein 3,NLRP3)、凋亡相关斑点样蛋白(apoptosis-associated speck-like protein containing a CARD,ASC)、裂解半胱氨酸天冬氨酸蛋白酶1(cleaved-caspasel)表达.鬼笔环肱(phalloidin)染色观察 HPC 骨架变化.2',7'-二氯二氢荧光素二乙酸酯(2',7'-dichlorodihydrofluorescein diacetate,DCFH-DA)检测细胞活性氧(reactive oxygen species,ROS)水平.酶联免疫吸附测定法(enzyme-linked immunosorbent assay,ELISA)检测细胞上清中炎性因子白细胞介素(interleukin,IL)-18、IL-1 β含量.结果 (1)与NG组比较,HG组MAGI2、nephrin表达水平较低(均P＜0.01),SGLT2表达较高(P＜0.01);足细胞形态变化及细胞骨架重构显著;细胞ROS水平较高(P＜0.01);NLRP3、ASC、cleaved-caspase1表达较高(均P＜0.01);细胞上清中IL-18、IL-1β浓度较高(均P＜0.05).(2)与HG组比较,Cana组MAGI2、nephrin表达升高(均P＜0.01);细胞形态变化及细胞骨架重构减轻;SGLT2表达下调(P＜0.05);细胞ROS水平下调;NLRP3、ASC、cleaved-caspase1 表达下调(P＜0.01);上清中 IL-18、IL-1β浓度下调(均P＜0.05).结论 卡格列净能有效减轻高糖诱导的HPC损伤及炎性反应,其机制可能与抑制ROS/NLRP3信号通路相关.
Abstract
Objective To investigate the efficacy and mechanism of canagliflozin(Cana)in the treatment of high glucose-induced human podocyte(HPC)injury.Methods The HPCs were divided into 5 groups:normal glucose group(NG group),mannitol group(MA group),high glucose group(HG group),Cana low dose(0.3 μmol/L)group and Cana high dose(1.0 μmol/L)group.Western blotting was used to examine the protein expressions of membrane-associated guanylate kinase inverted-2(MAG12),podocyte-associated protein nephrin,sodium-glucose transporter 2(SGLT2),NOD-like receptor thermal protein domain associated protein 3(NLRP3),apoptosis-associated speck-like protein containing a CARD(ASC),and cleaved-caspase1 in podocytes.Phalloidin staining of F-actin in podocytes was used to observe cytoskeletal injury.Intracellular reactive oxygen species(ROS)level of HPC was detected by the 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)probe.Levels of interleukin(IL)-18 and IL-1β in culture medium of podocytes were detected by enzyme-linked immunosorbent assay(ELISA).Results(1)Compared with the NG group,the protein expressions of MAGI2 and nephrin decreased(both P＜0.01),the protein expression of SGLT2 increased(P＜0.01),the changes of cell morphology and cytoskeleton remodeling were obvious,intracellular ROS level increased(P＜0.01),while NLRP3,ASC and cleaved-caspase1 protein expressions decreased in the HG group(all P＜0.01).The results of ELISA showed that IL-18 and IL-1β concentrations were higher in the HG group(both P＜0.05).(2)Compared with the HG group,in the Cana groups,MAGI2 and nephrin expressions up-regulated(both P＜0.01),the changes of cell morphology and cytoskeleton remodeling were alleviated.Meanwhile the Cana groups showed decreased SGLT2 expression(P＜0.05),lower ROS level,down-regulated NLRP3,ASC,cleaved-caspase1 expressions(all P＜0.01),and decreased concentrations of IL-18 and IL-1β in culture medium of podocytes(both P＜0.05).Conclusion Cana can improve high glucose-induced injury and inflammation in human podocyte,possibly due to the repression of the ROS/NLRP3 signaling pathway.
关键词
坎格列嗪/血糖/足细胞/炎症/活性氧/NLR家族,热蛋白结构域包含蛋白3Key words
Canagliflozin/Blood glucose/Podocytes/Inflammation/Reactive oxygen species/NLR family,pyrin domain-containing 3 protein引用本文复制引用
基金项目
国家自然科学基金青年科学基金(81803921)
上海市医学重点专科建设项目(ZK2019A12)
上海市普陀区卫生健康系统临床特色专科建设项目(2021tszk02)
出版年
2024
NETL
NSTL
万方数据