雷公藤甲素对PAN致足细胞损伤中mTOR/p70S6K/4EBP1信号通路的影响  

Effects of Triptolide on the Level of mTOR/p70S6K/4EBP1 Pathway in PAN Cultured Podocytes

作　　者：刘丽秋[1] 任帆[1] 韩润鸿 赵娜[1] 

机构地区：[1]青岛大学医学院附属医院肾病内科,青岛266003

出　　处：《中国中西医结合肾病杂志》2016年第2期110-113,共4页Chinese Journal of Integrated Traditional and Western Nephrology

基　　金：青岛市科技局基金资助项目(No.11-2-3-1-(2)-nsh)

摘　　要：目的:探讨雷公藤甲素(triptolide,TP)对m TOR/p70S6K/4EBP1信号通路的影响及其保护足细胞的可能机制。方法:氨基核苷嘌呤霉素(puromycin aminonucleoside,PAN)刺激体外培养的足细胞24 h建立模型。细胞增殖检测试剂盒(CCK-8法)检测不同浓度雷公藤作用下PAN损伤足细胞的细胞存活率。后将足细胞随机分组:(1)对照组;(2)PAN组(PAN组,PAN 50μg/ml);(3)TP组(PAN 50μg/ml+TP 10 ng/ml),予以相应刺激24 h。采用实时荧光定量PCR检测足细胞m TOR、4EBP1、p70S6K、Akt、synaptopodin mRNA表达量,western blot检测足细胞p-mTOR、p-4EBP1、p-p70S6K、p-Akt、synaptopodin蛋白表达量。结果:(1)CCK-8结果显示,PAN造成足细胞损伤时加入浓度为3、10 ng/ml雷公藤甲素浓度均使足细胞活性上升,10 ng/ml时作用最明显(均P<0.05)。(2)PAN增加m TOR、4EBP1、p70S6K、Akt mRNA及蛋白的表达量,减少synaptopodin mRNA及蛋白表达水平(均P<0.05)。(3)雷公藤甲素共处理细胞后,m TOR、4EBP1、p70S6K、Akt mRNA及蛋白的表达水平明显下降,而synaptopodin mRNA及蛋白表达水平明显升高(均P<0.05)。结论:雷公藤甲素可能通过抑制m TOR/p70S6K/4EBP1信号通路,减轻足细胞损伤。Objective： This article aims to explore the influence of triptolide on m TOR / p70S6 K /4EBP1 pathway and its possible mechanism to protect podocytes. Methods： Cultured mice podocytes cell 5（ MPC5） in vitro were coincubated with PAN for 24 h. We added different concentrations of triptolide into PAN cultured podocytes and then detected the cell activity of each goup by CCK8 assay. Then podocytes were randomly divided into four groups：（ 1） Control group;（ 2） PAN group（ PAN 50 μg / ml）; PAN group（ PAN 50 μg / ml）;（ 3） PAN ＋ Triptolide group（ PAN 50 μg / ml ＋ TP 10 ng / ml）. Cultivating time of each group is 24 h. The mRNA expressions of m TOR、4EBP1、p70S6K、Akt and synaptopodin in podocyte were detected by real- time quantitative RT- PCR and the protein expressions of p- m TOR、p- 4EBP1、p- p70S6K、p- Akt and synaptopodin were detected by westernblot. Results：The results of CCK- 8 showed that adding 3,10 ng / ml triptolide increased the cells viability and the concentration of 10 ng / ml was the most obvious（ P 0. 05）. Compared with the control group,the expressions of m TOR,4EBP1,p70S6 K and Akt were up- regulated considerably while the expression of synaptopodin was significantly decreased（ P 0. 05） detected by RT- PCR and westernblot. Compared with PAN group,triptolide alleviated the up- regulation of m TOR,4EBP1,p70S6 K and Akt expressions and down-regulation of synaptopodin expression（ P 0. 05） detected by RT- PCR and westernblot. Conclusion： Triptolide can protect podocytes through alleviating the activation of m TOR / p70S6K /4EBP1 pathway in PAN cultured podocytes.

关 键 词：雷公藤甲素 足细胞损伤 PAN mTOR/p70S6K/4EBP1 

分 类 号：R285[医药卫生—中药学]