天津医药

天津医药 ›› 2020, Vol. 48 ›› Issue (3): 161-164.doi: 10.11958/20193415

• 细胞与分子生物学 •    下一篇

大黄素对 TGF-β1诱导的人肾小管上皮细胞上皮间质转分化及 Mfn2表达的影响

龙脉，李梦婕，柯本，黄金菁，房向东   

  1. 南昌大学第二附属医院肾脏内科（邮编 330000）
  • 收稿日期:2019-11-11 修回日期:2020-01-31 出版日期:2020-03-15 发布日期:2020-04-11
  • 通讯作者: 龙脉 E-mail:longmdoc@163.com

Effects of emodin on TGF-β1-induced epithelial-mesenchymal transition and expression of Mfn2 in human renal tubular epithelial cells

LONG Mai,LI Meng-jie,KE Ben,HUANG Jin-jing,FANG Xiang-dong   

  1. Department of Nephrology, the Second Affiliated Hospital of Nanchang University, Nanchang 330000, China
  • Received:2019-11-11 Revised:2020-01-31 Published:2020-03-15 Online:2020-04-11

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摘要： 目的 探讨大黄素（EM）对转化生长因子 β1（TGF-β1）诱导的人肾小管上皮细胞（HK-2）上皮间质转分化及线粒体融合蛋白 2（Mfn2）的影响。方法 采用 CCK-8法检测不同浓度 EM对 HK-2细胞增殖的影响；实验分空白对照组、TGF-β1组、TGF-β1+EM（10、20、40 μmol/L）组，EM干预组予以不同浓度的大黄素处理 HK-2细胞 30 min后，TGF-β1组和 EM干预组同时给予浓度为 10 μg/L的 TGF-β1刺激 48 h收集细胞，Western blot法检测 α-平滑肌肌动蛋白（a-SMA）、E-钙黏蛋白（E-cadherin）、结缔组织生长因子（CTGF）及 Mfn2蛋白的表达。结果 CCK-8结果提示 0、10、20、40、80、160 μmol/L 浓度的 EM 处理组的 HK-2 细胞存活率（%）分别为 100.00±0.00、90.89±2.17、88.52±1.67、87.72±2.00、63.46±1.73、52.19±2.60 ，呈逐渐降低趋势（P＜0.01）；Western blot结果表明，与空白对照组比较，TGF-β1刺激后，HK-2细胞中 α-SMA、CTGF及 Mfn2蛋白水平升高，E-cadherin蛋白表达降低（P＜0.05），与 TGF-β1组比较，给予不同浓度大黄素干预后，α-SMA、CTGF及 Mfn2蛋白水平降低，E-cadherin蛋白表达升高（P＜0.05），且呈一定剂量依赖性。结论 大黄素缓解了 TGF-β1诱导的 HK-2细胞上皮间质转分化，可能通过减少 TGF-β1诱导的 Mfn2的表达来减缓肾脏纤维化。

关键词: 大黄素, 转化生长因子 β1, 上皮间质转分化, 线粒体融合蛋白 2, 肾脏纤维化

Abstract: Objective To investigate the effect and mechanism of emodin (EM) on TGF- β1-induced epithelialmesenchymal transition and the expression of mitofusin 2 (Mfn2) in human renal tubular epithelial cells (HK-2). Methods CCK-8 was used to detect the effects of different concentrations of emodin on the proliferation of HK-2 cells. The experiment cells were divided into blank control group, TGF-β1 group and TGF-β1+EM group. The emodin intervention group was treated with different concentrations of emodin (10, 20 and 40 μmol/L) for 30 min, and the TGF-β1 group and the emodin intervention group were simultaneously stimulated with TGF- β1 at a concentration of 10 μg/L for 48 h. The cells were then harvested and the expressions of α-smooth muscle actin (α-SMA), E-cadherin, connective tissue growth factor (CTGF) and Mfn2 proteins were detected by Western blot assay. Results The CCK-8 result suggested that the survival rate(% ) of HK-2 cells were 100.00±0.00, 90.89±2.17, 88.52±1.67, 87.72±2.00, 63.46±1.73 and 52.19±2.60 in the EM treatment groups at 0, 10, 20, 40, 80, and 160 μmol/L concentrations, showing a gradually decreasing trend (P＜0.01).Western blot results showed that the protein levels of α-SMA, CTGF and Mfn2 increased in HK-2 cells, and the expression of E-cadherin protein decreased after stimulation with TGF-β1 compared with those of the blank control group (P＜0.05).Compared with the TGF- β1 group, α -SMA, CTGF and Mfn2 protein levels were decreased, and E-cadherin protein expression was increased after treatment with different concentrations of emodin (P＜0.05) in a dose-dependent manner. Conclusion Emodin attenuates TGF- β1-induced epithelial-mesenchymal transition in human renal tubular epithelial cells, which may ameliorate renal fibrosis by reducing the expression of TGF-β1-induced Mfn2.

Key words: emodin, transforming growth factor β1, epithelial-mesenchymal transition, mitofusin2, renal fibrosis