卡格列净通过抑制TXNIP信号改善肾小管上皮细胞氧化应激损伤的机制研究OA
Mechanism of canagliflozin on ameliorating oxidative stress injury in renal tubular epithelial cells by inhibiting TXNIP signaling
目的:观察卡格列净(canagliflozin,Cana)对高糖诱导肾小管上皮细胞HK-2氧化应激损伤的疗效,并探讨其相关机制.方法:CCK-8法检测Cana对HK-2增殖的影响以筛选实验安全浓度.将HK-2分为6 组:正常组(NG组)、甘露醇组(MA组)、高糖组(HG组)及Cana低、中、高(1、3、10 μmol/L)剂量组.Western blot检测TXNIP、NLRP3、DRP1、MFN1和MFN2蛋白表达,试剂盒检测细胞中GSH-PX、SOD酶活性和MDA水平,ELISA法检测细胞上清中炎症因子IL-1β和IL-18的含量,mitoSOX探针检测细胞内线粒体活性氧水平,免疫荧光检测DRP1在线粒体的累积.转染shRNA沉默TXNIP,观察对NLRP3、DRP1、MFN1和MFN2蛋白表达的影响.结果:与HG组比较,Cana可抑制高糖诱导的氧化应激和炎症损伤,表现为GSH-PX和SOD酶活性升高(P<0.05),MDA含量减少(P<0.05),细胞上清中炎症因子IL-1β和IL-18的含量降低(P<0.05);还可抑制TXNIP及NLRP3蛋白表达(P<0.05);此外Cana在减少线粒体中活性氧累积同时,可下调线粒体分裂蛋白DRP1表达(P<0.05),并上调线粒体融合蛋白MFN1和MFN2表达(P<0.05).沉默TXNIP后,Cana对NLRP3、DRP1、MFN1和MFN2的干预效应被明显削弱.结论:Cana可减轻高糖诱导的肾小管上皮细胞氧化应激损伤,其机制与TXNIP信号通路有关.
Objective:To observe the efficacy of canagliflozin(Cana)on treating high glucose-induced oxidative stress injury in renal tubular epithelial cells HK-2 and to explore the related mechanism.Methods:The effect of Cana on the proliferation of HK-2 was detected by CCK-8 for screening the experimental safe concentration HK-2 was divided into 6 groups:the normal group(NG group),the mannitol group(MA group),the high glucose group(HG group),the Cana low-dose group,the Cana medium-dose group,and the Cana high-dose group(1,3,10 μmol/L).TXNIP,NLRP3,DRP1,MFN1,and MFN2 protein ex-pressions were detected by Western blot.Intracellular GSH-PX and SOD enzyme activities,and MDA level were examined by kit assays.The contents of inflammatory factors IL-1β and IL-18 in the cell supernatant were detected by ELISA.The level of intra-cellular mitochondrial reactive oxygen species was detected by mitoSOX Red probe,and the accumulation of DRP1 in mitochon-dria was detected by immunofluorescence.TXNIP was silenced by shRNA transfection to investigate its effects on the protein ex-pression levels of NLRP3,DRP1,MFN1,and MFN2.Results:Compared to the HG group,Cana could inhibit high glucose-induced oxidative stress and inflammatory injury,as evidenced by increased GSH-PX and SOD enzyme activity(P<0.05),reduced MDA content(P<0.05),and decreased the levels of inflammatory factors IL-1β and IL-18 in the cell supernatant(P<0.05).Cana could also inhibit the expressions of TXNIP and NLRP3 proteins(P<0.05).Additionally,while reducing the accumulation of reactive oxygen species in mitochondria,Cana could downregulate the expression of mitochondrial fission protein DRP1(P<0.05)and upregulate the expression of mitochondrial fusion proteins MFN1 and MFN2(P<0.05).After TXNIP gene silencing,the intervention effect of Cana on NLRP3,DRP1,MFN1,and MFN2 was significantly weakened.Conclusion:Cana can alleviate the oxidative stress injury of renal tubular epithelial cells induced by high glucose,and the mechanism is related to TXNIP signaling pathway.
周宇颖;王利;王浩
上海中医药大学附属普陀医院肾内科,上海 200062上海中医药大学附属普陀医院肾内科,上海 200062上海中医药大学附属普陀医院肾内科,上海 200062
医药卫生
卡格列净TXNIP肾小管上皮细胞氧化应激
CanagliflozinTXNIPRenal tubular epithelial cellsOxidative stress
《海南医科大学学报》 2026 (4)
241-247,7
This study was supported by the Clinical Specialty of Shanghai Putuo District Health System(2021tszk02) 上海市普陀区卫生健康系统临床特色专科建设项目(2021tszk02)
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