芪地糖肾方调控NLRP3/Caspase-1/GSDMD通路介导高糖刺激肾小球内皮细胞焦亡的研究  

Qi Di Tang Shen decoction mediates high glucose-stimulated glomerular endothelialcells’ pyroptosis via regulating NLRP3/Caspase1/GSDMD pathway

作　　者：史扬 董昭熙 周盈 谢惠迪 郭燕 宿家铭 郑毅成 柳红芳[1] SHI Yang;DONG Zhaoxi;ZHOU Ying;XIE Huidi;GUO Yan;SU Jiaming;ZHENG Yicheng;LIU Hongfang(Dongzhimen Hospital of Beijing University of Chinese Medicine,Beijing 100700,China;Beijing Puren Hospital,Beijing 100062,China;First Medical Center of Chinese PLA General Hospital,State Key Laboratory of Kidney Diseases,Beijing 100853,China)

机构地区：[1]北京中医药大学东直门医院,北京100700 [2]北京市普仁医院,北京100062 [3]解放军总医院第一医学中心肾脏病全国重点实验室,北京100853

出　　处：《现代中西医结合杂志》2024年第1期1-7,共7页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：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)。

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100μg/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。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 medium,30 mmol/L high-glucose medium+100μg/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 detected by cellular immunofluorescence staining.Results Compared with the normal group,the relative expressions of NLRP3,Caspase1,GSDMD,ASC,IL-18 and IL-1βin 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-1βin 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.

关 键 词：糖尿病肾脏病 肾小球内皮细胞 芪地糖肾方 焦亡 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 

分 类 号：R-332[医药卫生]