文章摘要

雷公藤甲素通过抑制TLR4基因对糖尿病肾病足细胞的保护作用

作者: 1李 乔, 1张 博
1 中国中医科学院广安门医院南区内分泌科,北京 102618
通讯: 张 博 Email: hepg29@sina.com
DOI: 10.3978/j.issn.2095-6959.2020.04.004

摘要

目的:研究雷公藤甲素对糖尿病肾病足细胞的保护作用及作用机制。方法:体外培养永生化小鼠肾小球足细胞,以高糖刺激足细胞诱导足细胞损伤,雷公藤甲素进行干预,采用qRT-PCR技术和蛋白质印迹法检测雷公藤甲素对高糖诱导的足细胞中Toll样小体4(Toll-like receptor 4,TLR4)表达水平的影响,采用慢病毒感染过表达TLR4,qRT-PCR和蛋白质印迹法检测感染效果,噻唑蓝比色法(MTT)检测各组足细胞增殖能力,流式细胞仪检测各组足细胞凋亡情况;酶联免疫吸附法(ELISA)检测各组足细胞培养上清液中IL-6和TNF-α的含量。结果:qRT-PCR和蛋白质印迹法检测结果显示雷公藤甲素下调高糖诱导的足细胞中TLR4的表达;MTT结果显示雷公藤甲素增加高糖诱导的足细胞的增殖活力;流式细胞术检测结果显示雷公藤甲素抑制高糖诱导的足细胞凋亡;ELISA检测结果显示雷公藤甲素抑制高糖诱导的足细胞分泌IL-6和TNF-α;过表达TLR4逆转雷公藤甲素对高糖诱导的足细胞保护作用。结论:雷公藤甲素能够通过抑制TLR4对糖尿病肾病足细胞起保护作用。
关键词: 雷公藤甲素;Toll样小体4;糖尿病肾病;足细胞

Protective effect of triptolide on podocytes of diabetic nephropathy by inhibiting TLR4 gene

Authors: 1LI Qiao, 1Zhang Bo
1 Department of Endocrine, South District of Guang’anmen Hospital of China Academy of Chinese Medical Sciences, Beijing 102618, China

CorrespondingAuthor:Zhang Bo Email: hepg29@sina.com

Abstract

Objective: To investigate the protective effect and mechanism of triptolide on diabetic nephropathy podocytes. Methods: The immortalized glomerular podocytes of mice were cultured in vitro, and stimulating podocytes to induce podocyte injury with high glucose. Triptolide was intervened by real-time quantitative PCR (qRT-PCR) and Western blotting. The effect of triptolide on the expression of Toll-like receptor 4 (TLR4) in podocytes induced by high glucose were detected by qRT-PCR and Western blotting. The overexpressed TLR4 were infected by lentivirus, and the infection effect was detected by qRT-PCR and Western blotting. The proliferation of podocytes in each group was detected by MTT. Flow cytometry was used to detect the apoptosis of podocytes in each group. The levels of IL-6 and TNF-α in the culture supernatant of podocytes of each group were detected by ELISA. Results: qRT-PCR and Western blotting showed that triptolide down-regulated the expression of TLR4 in podocytes induced by high glucose; MTT results showed that triptolide increased the proliferation of podocytes induced by high glucose; the results of cytometry showed that triptolide inhibited the apoptosis of podocytes induced by high glucose; ELISA results showed that triptolide inhibited the secretion of IL-6 and TNF-α by podocytes induced by high glucose; overexpression of TLR4 reversed the protective effect of triptolide on high glucose-induced podocytes. Conclusion: Triptolide can protect the podocytes of diabetic nephropathy by inhibiting TLR4.
Keywords: triptolide; Toll-like receptor 4; diabetic nephropathy; podocyte