姜黄素抑制TGF-β1诱导的人肾小管上皮细胞系表型转化

基础医学与临床 ›› 2020, Vol. 40 ›› Issue (1): 48-53.

姜黄素抑制TGF-β1诱导的人肾小管上皮细胞系表型转化

陈民佳¹, 杜娟¹, 袁丹凤¹, 张蜀¹, 黄宏¹, 朱方强^2*

1.陆军军医大学大坪医院创伤、烧伤与复合伤国家重点实验室,重庆 400042;
2.重庆医科大学第三附属医院泌尿外科,重庆 401120

收稿日期:2019-06-06 修回日期:2019-11-12 出版日期:2020-01-05 发布日期:2019-12-27
通讯作者: ^*zfq68757079@sina.com
基金资助:
国家自然科学基金(81571912,81372059);重庆市院士牵头科技创新引导专项(cstc2017zdcy-yszxX0002);创伤、烧伤与复合伤国家重点实验室开放基金(200717);重庆高校创新团队建设计划资助项目(CXTDX201601005)

Curcumin inhibits TGF-β1-induced phenotypic transition of human kidney tubular cells (HKCs)

CHEN Min-jia¹, DU Juan¹, YUAN Dan-feng¹, ZHANG Shu¹, HUANG Hong¹, ZHU Fang-qiang^2*

1. State Key Laboratory of Trauma, Burn and Combined Injury, Daping Hospital, Army Medical University, Chongqing 400042;
2. Department of Urology, the Third Affiliated Hospital of Chongqing Medical University, Chongqing 401120, China

Received:2019-06-06 Revised:2019-11-12 Online:2020-01-05 Published:2019-12-27
Contact: ^*zfq68757079@sina.com

摘要/Abstract

摘要： 目的探讨姜黄素抑制TGF-β1诱导的肾纤维化的作用及分子生物学机制。方法应用10 ng/mL TGF-β1单独或联合姜黄素(Cur)(6.25、12.5、25、50和100 μmol/L)共同作用人肾小管上皮细胞系(HKCs)72 h,倒置相差显微镜下观察细胞形态,免疫组化检测纤维细胞特异蛋白-1 (FSP-1)表达;同时,通过Western blot检测肾小管上皮细胞标志物E-cardhrin和细胞角化蛋白以及基质标志物波形蛋白、α-平滑肌肌动蛋白(α-SMA)和Ⅰ型胶原的表达变化,以及AKT/mTOR信号通路关键蛋白的表达。结果 HKCs贴壁增殖,呈铺路石样外观。TGF-β1单独刺激能明显促进细胞由上皮样形态向纤维样细胞形态转化。当TGF-β1与姜黄素联合作用于HKCs时,姜黄素在12.5～50 μmol/L浓度时能明显对抗TGF-β1诱导的细胞形态转变而维持HKCs细胞铺路石样外观,同时,TGF-β1能明显抑制HKCs细胞E-cardhrin和角化蛋白基因表达(P＜0.05,P＜0.01),而促进FSP1阳性细胞显著增加(P＜0.05,P＜0.01);上调波形蛋白、α-SMA和I型胶原的表达(P＜0.05,P＜0.01);并且,AKT/mTOR信号通路关键蛋白Akt、mTOR、p70S6K、4E-BP1和eIF4E的磷酸化水平表达也显著增加(P＜0.05,P＜0.01)。而姜黄素能抑制TGF-β1诱导的HKCs表型转化,同时下调TGF-β1诱导AKT/mTOR信号通路关键蛋白的磷酸化水平(P＜0.05)。结论姜黄素通过抑制Akt/mTOR信号通路对抗TGF-β1诱导的上皮细胞-间质细胞转化,可能是姜黄素抗肾纤维化的关键分子机制之一。

关键词: 姜黄素, 肾纤维化, 上皮-间质细胞转化, TGF-β1, Akt/mTOR信号通路

Abstract: Objective To study the effects of curcumine on renal tubular epithelial-mesenchymal transition (EMT) and its possible mechanisms. Methods The effect of curcumin on the morphology of human kidney cells (HKCs) in the presence of TGF-β1 alone or in combination with different concentration curcumine was observed by phase contrast microscopy. The expressions of E-cardhrin, keratin, vimentin, alpha smooth muscle actin (α-SMA),fibroblast-specific protein 1 (FSP1) and key proteins of AKT/mTOR pathway were analyzed by immunohistochemical stain and Western blot. Results HKC cells showed a classic cobblestone morphology. Exposure of HKCs to TGF-β1 for 72 h induced a complete conversion of the epithelial cell to myofibroblast. When HKCs were co-incubated with TGF-β1 and various concentrations of curcumin for 72 h, curcumine maintained the epithelial morphology in a dose-dependent manner (from 12.5 to 50 μg/mL), antagonizing TGF-β1-induced the down-regulation expression of E-cardhrin, keratin and the up-rgulation of FSP1, vimentin and α-SMA. Importantly. Curcumine effectively suppressed the activity of the Akt/mTOR pathway in HKCs as demonstrated by a remarkable reduction of the Akt, mTOR, p70S6K, 4E-BP1和eIF4E phosphorylation. Conclusions Curcumine is a potent inhibitor of TGF-β1-induced epithelial-mesenchymal transition (EMT) and antagonizes TGF-β1-driven fibrogenesis through inhibition of the activity of the Akt/mTOR pathway.

Key words: curcumine, renal fibrosis, epithelialto-mesenchymal transition (EMT), TGF-β1, AKt/mTOR pathway

中图分类号:

R363.2

陈民佳, 杜娟, 袁丹凤, 张蜀, 黄宏, 朱方强. 姜黄素抑制TGF-β1诱导的人肾小管上皮细胞系表型转化[J]. 基础医学与临床, 2020, 40(1): 48-53.

CHEN Min-jia, DU Juan, YUAN Dan-feng, ZHANG Shu, HUANG Hong, ZHU Fang-qiang. Curcumin inhibits TGF-β1-induced phenotypic transition of human kidney tubular cells (HKCs)[J]. Basic & Clinical Medicine, 2020, 40(1): 48-53.

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