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

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(0.3μmol/L)组、Cana高剂量(1.0μmol/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-caspase1)表达。鬼笔环肽(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-1β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。

Canagliflozin attenuates human podocyte injury through inhibiting reactive oxygen species/NOD-like receptor thermal protein domain associated protein 3 signaling pathway

Objective To investigate the efficacy and mechanism of canagliflozin (Cana) in the treatment of high glucoseinduced 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(0.3μmol/L) group and Cana high dose (1.0μmol/L) group.