摘要
目的 探究芪地糖肾方通过NOD样受体家族热蛋白结构域相关蛋白3/半胱氨酸天冬氨酸蛋白酶-1/消皮素D蛋白(NLRP3/Caspase-1/GSDMD)通路介导高糖刺激的肾小球内皮细胞焦亡的作用.方法 将肾小球内皮细胞分为正常组、高糖组、芪地糖肾方组,分别予完全培养基、30 mmol/L高糖培养基、30 mmol/L高糖培养基+芪地糖肾方100 μg/mL培养48 h,采用细胞免疫荧光染色法观察细胞中NLRP3 表达情况及细胞骨架情况,采用Western blot法检测细胞中NOD样受体热蛋白结构域相关蛋白3(NLRP3)、半胱氨酸天冬氨酸蛋白酶1(Caspase-1)、消皮素D蛋白(GSDMD)、凋亡相关斑点样蛋白(ASC)、白细胞介素-1β(IL-1β)、白细胞介素-18(IL-18)表达情况.结果 与正常组比较,高糖组细胞中NLRP3、Caspase-1、GSDMD、ASC、IL-18、IL-1β蛋白相对表达量均明显升高(P均＜0.05),NLRP3 荧光表达信号增强,细胞骨架损伤明显;与高糖组比较,芪地糖肾方组NLRP3、Caspase-1、GSDMD、ASC、IL-18、IL-1β蛋白相对表达量均明显降低(P均＜0.05),NLRP3 荧光表达信号减弱,细胞骨架损伤状态改善.结论 芪地糖肾方可通过抑制NLRP3/Caspase-1/GSDMD焦亡相关通路激活改善高糖诱导的肾小球内皮细胞损伤.
Abstract
Objective It is to investigate the effect of Qi Di Tang Shen decoction on high glucose-stimulated glomerular endothelial cells(mGECs)via Nod-like receptor protein 3/cystein-asparate protease-1/gasdermin D(NLRP3/Caspase1/GSDMD)pathway.Methods The mGECs were divided into three groups,named as normal group,high glucose group and Qi Di Tang Shen decoction group,which were respectively cultured in complete medium,30 mmol/L high-glucose medi-um,30 mmol/L high-glucose medium +100 μg/mL Qi Di Tang Shen decoction for 48 h.The expression of NLRP3 in cells and cytoskeleton were observed by cellular immunofluorescence staining,the expressions of NLRP3,Caspase1,apoptosis-associated spot-like protein(ASC),GSDMD,interleukin-1β(IL-1β)and interleukin-18(IL-18)in mGECs were detec-ted by cellular immunofluorescence staining.Results Compared with the normal group,the relative expressions of NLRP3,Caspase1,GSDMD,ASC,IL-18 and IL-1β in mGECs of the high glucose group were significantly increased(all P＜0.05),the fluorescent expression of NLRP3 was enhanced,and the cytoskeleton damage was obvious.Compared with the high glucose group,the relative expressions of NLRP3,Caspase1,GSDMD,ASC,IL-18 and IL-1β in Qi Di Tang Shen decoction group were significantly down-regulated(all P＜0.05),and the fluorescent expression of NLRP3 was decreased and the cytoskeletal damage was improved.Conclusion Qi Di Tang Shen decoction can improve the high-glucose induced injury of glomerular endothelial cells by inhibiting the activation of NLRP3/Caspase1/GSDMD pathway.
基金项目
北京市自然科学基金资助项目(7212180)
国家自然科学基金资助项目(82004297)
国家中医药管理局国家中医临床研究基地业务建设科研专项课题(JDZX2015305)
维普
万方数据