文献类型：期刊文献

中文题名：基于PTEN/PI3K/Akt/mTOR信号通路探讨淫羊藿苷联合地塞米松对阿霉素诱导足细胞损伤的保护作用及机制

英文题名：Protective Effect and Mechanism of Icariin Combined with Dexamethasone on Doxorubicin-Induced Podocyte Injury Based on PTEN/PI3K/Akt/mTOR

作者：吕娟[1,2];张云霞[1,2];白俊嫄[1];蒲晓薇[1];戴恩来[1]

第一作者：吕娟

机构：[1]甘肃中医药大学,甘肃兰州730000;[2]甘肃省中医院,甘肃兰州730050

第一机构：甘肃中医药大学

年份：2023

卷号：34

期号：6

起止页码：739

中文期刊名：中药新药与临床药理

外文期刊名：Traditional Chinese Drug Research and Clinical Pharmacology

收录：CSTPCD;;北大核心:【北大核心2020】；CSCD:【CSCD2023_2024】；

基金：国家自然科学基金委员会地区科学基金项目(81760799)

语种：中文

中文关键词：阿霉素;足细胞损伤;地塞米松;淫羊藿苷;PTEN/PI3K/Akt/mTOR信号通路;自噬

外文关键词：doxorubicin;podocyte injury;Dexamethasone;icariin;PTEN/PI3K/AKt/mTOR signaling pathway;autophagy

摘要：目的探讨淫羊藿苷联合地塞米松对阿霉素诱导足细胞损伤的保护作用及分子机制。方法体外培养大鼠肾小球足细胞,阿霉素诱导建立足细胞损伤模型,分为:空白组、模型组、淫羊藿苷联合地塞米松组及氯喹组。免疫荧光法测定足细胞特异性骨架蛋白synaptopodin的表达;转染mRFP-GFP-LC3双荧光自噬指示体系,激光共聚焦显微镜观察足细胞内自噬流的强弱;透射电镜观察足细胞内自噬体、自噬溶酶体的变化;Western Blot法检测足细胞自噬相关蛋白LC3-Ⅱ、p62及自噬信号通路蛋白PTEN、p-PI3K、p-Akt、p-mTOR的表达。结果与模型组比,淫羊藿苷联合地塞米松组的synaptopodin的表达升高(P<0.05);足细胞内自噬流水平、自噬体及自噬溶酶体数量均增加(P<0.01);LC3-Ⅱ、PTEN的表达上调(P<0.01),但p62、pPI3K、p-Akt、p-mTOR的表达下调(P<0.01)。与淫羊藿苷联合地塞米松组比,氯喹组synaptopodin的表达降低(P<0.05);足细胞内自噬流的水平降低;自噬体数量增加(P<0.05),自噬溶酶体的数量减少(P<0.01);且LC3-Ⅱ和p62表达同步升高(P<0.05,P<0.01),但PTEN、p-PI3K、p-Akt、p-mTOR的表达无明显变化(P>0.05)。结论淫羊藿苷联合地塞米松可能通过激活PTEN,抑制PI3K/Akt/mTOR信号通路,增强足细胞自噬水平,达到保护足细胞的作用。
Objective To investigate the protective effect of icariin combined with Dexamethasone on doxorubicininduced podocyte injury and its molecular mechanism.Methods Rat glomerular podocytes were cultured in vitro and induced with doxorubicin to establish podocyte injury model.The cells were divided into blank group,model group,drug treatment group(icariin combined with Dexamethasone)and chloroquine group.The expression of synaptopodin(a podocyte specific protein)was detected by immunofluorescence assay.The mRFP-GFP-LC3 dual fluorescence autophagy indicator system was used to observe the intensity of autophagy flow in podocytes with laser confocal microscope.The changes of autophagosomes and autophagolysosomes in podocytes were observed by transmission electron microscopy.The expressions of autophagy related proteins(LC3-Ⅱ,p62)and autophagy signaling pathway proteins(PTEN,p-PI3K,p-Akt and p-mTOR)in podocyte were detected by Western Blot.Results Compared with the model group,the expression of synaptopodin in the drug treatment group was significantly increased(P<0.05).The level of autophagic flow,the number of autophagosomes and autophagosomes in podocytes were increased(P<0.01).The expressions of LC3-Ⅱand PTEN were up-regulated(P<0.01),but the expressions of p62,p-PI3K,p-Akt and p-mTOR were down-regulated(P<0.01).Compared with the drug treatment group,the expression of synaptopodin in the chloroquine group was decreased(P<0.05).The level of autophagy flow in podocytes was found to be decreased.However,the number of autophagosomes increased(P<0.05),while the number of autophagosomes decreased(P<0.01).The expressions of LC3-II and P62 were increased simultaneously(P<0.05,P<0.01),but the expressions of PTEN,p-PI3K,p-Akt and p-mTOR did not change significantly(P>0.05).Conclusion Icariin combined with Dexamethasone may activate PTEN,inhibit PI3K/Akt/mTOR signaling pathway,enhance the level of podocyte autophagy,so as to achieve the role of podocyte protection.