大黄素通过激活自噬对TGF-β1诱导的HK-2细胞纤维化因子的影响  

Effect of emodin on fibrosis factor of HK-2 cells stimulated by TGF-β1 through activation of autophagy

作　　者：窦芳 丁一 姚敏娜 简宇凡 王文军 赵先 王婧雯 文爱东 DOU Fang;DING Yi;YAO Min-na;JIAN Yu-fan;WANG Wen-jun;ZHAO Xian;WANG Jing-wen;WEN Ai-dong(Dept of Pharmacy,Xijing Hospital,the Fourth Military Medical University,Xi’an 710032,China)

机构地区：[1]空军军医大学西京医院药剂科,陕西西安710032

出　　处：《中国药理学通报》2018年第11期1555-1559,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81603385);陕西省自然科学基金资助项目(No 2017JM8006)

摘　　要：目的研究大黄素对TGF-β1刺激的人肾小管上皮细胞(HK-2)损伤的保护作用,并探讨其可能的机制。方法 TGF-β1刺激HK-2细胞24 h后,收集细胞,用于免疫荧光、免疫印迹和实时荧光定量PCR(qPCR)分析。qPCR法分别检测肾纤维化蛋白TGF-β1、纤连蛋白(FN),以及自噬蛋白LC3、Beclin-1 mRNA的表达;Western blot法分别检测TGF-β1、Ⅰ型胶原蛋白(COLⅠ)、LC3和Beclin-1的蛋白表达;免疫荧光检测肾纤维化相关因子FN在HK-2细胞模型中的表达。结果 qPCR、Western blot和免疫荧光实验结果表明,与Control组比较,TGF-β1组细胞肾纤维化相关蛋白TGF-β1、COLⅠ、FN表达升高,自噬标志蛋白LC3和Beclin-1的表达也升高。qPCR、Western blot和免疫荧光实验结果表明,与TGF-β1组比较,TGF-β1+大黄素组肾纤维化相关蛋白TGF-β1、COLⅠ、FN表达降低,LC3和Beclin-1的表达仍然增加。结论大黄素干预后,促进TGF-β1细胞模型自噬增加,改善肾纤维化的程度,保护细胞模型损伤。Aim To investigate the role and mechanism of emodin in fibrosis of human renal tubular epithelial cells(HK-2)stimulated by TGF-β1.Methods After HK-2 cells were stimulated with TGF-β1 for 24 h,cells were collected for immunofluorescence,immunoblot and qPCR analysis.qPCR was used to detect TGF-β1,fibronectin,LC3 and Beclin-1.Western blot was employed to determine the protein expressions of TGF-β,collagen I,LC3 and Beclin-1.Immunofluorescence was used to detect fibronectin.Results The results of qPCR,Western blot and immunofluorescence showed that compared with control group,the expressions of TGF-β1,collagenⅠand fibronectin in TGF-β1 group significantly increased.Compared with those in TGF-β1 group,the expressions of TGF-β1,collagenⅠand fibronectin were reduced in TGF-β1+EM group.Additionally,the expressions of autophagy protein LC3 and Beclin-1 were up-regulated.Conclusion Emodin can promote autophagy,ameliorate renal interstitial fibrosis and protect cell damage.

关 键 词：慢性肾脏病 大黄素 纤维化 TGF-Β1 HK-2 自噬 

分 类 号：R284.1[医药卫生—中药学] R322.61[医药卫生—中医学]