窦 芳,丁 一,郭奇彦,王文军,简宇凡,赵 先,王婧雯,文爱东.大黄素对TGF-β1诱导的肾小管上皮细胞间质转分化的影响[J].现代生物医学进展英文版,2018,(21):4007-4011. |
大黄素对TGF-β1诱导的肾小管上皮细胞间质转分化的影响 |
To Study the Effect of Emodin on the Transdifferentiation of Human Renal Tubular Epithelial Cell Induced by TGF-β1 |
Received:July 10, 2018 Revised:July 30, 2018 |
DOI:10.13241/j.cnki.pmb.2018.21.002 |
中文关键词: 大黄素 上皮间质转分化 人肾小管上皮细胞 转化生长因子-β1 α-平滑肌肌动蛋白 |
英文关键词: Emodin Epithelial-mesenchymal transition Human renal tubular epithelial cell Transforming growth factor-β1 α-Smooth muscle actin |
基金项目:国家自然科学基金项目(81603385);陕西省自然科学基金项目(2017JM8006) |
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中文摘要: |
摘要 目的:探讨大黄素对TGF-β1诱导的人肾小管上皮细胞(HK-2)间质转分化的影响。方法:不同浓度大黄素分别作用于TGF-β1诱导HK-2细胞24 h和48 h,通过细胞增殖实验确定最佳大黄素最佳给药浓度。TGF-β1诱导HK-2细胞24 h后收集细胞用于免疫印迹Western blot和实时荧光定量PCR(RT-PCR)分析。Western印迹法分别检测纤维化相关蛋白Collagen IV的表达,和肾小管上皮细胞向间充质细胞转分化关键蛋白α-SMA和E-Cadherin的表达;RT-PCR法检测肾小管上皮细胞向间充质细胞转分化关键蛋白α-SMA的表达。结果:由细胞增殖实验结果表明40 μM大黄素是最佳给药浓度。Western结果表明,与模型组相比,大黄素组下调纤维化相关蛋白Collagen IV的表达,大黄素组与模型组蛋白差异有统计学意义(P<0.05)。与模型组相比,大黄素组下调α-SMA蛋白表达水平,而上调E-Cadherin蛋白表达,差异有统计学意义(P<0.05)。RT-PCR结果表明,与模型组相比,大黄素组降低α-SMA mRNA的含量,大黄素组与模型组α-SMA mRNA含量差异有统计学意义(P<0.05)。结论:大黄素可通过抑制TGF-β1诱导的HK-2细胞间质转分化,从而发挥延缓肾间质纤维化的过程。 |
英文摘要: |
ABSTRACT Objective: To study the effect of Emodin on the transdifferentiation of human renal tubular epithelial (HK-2) cell in- duced by TGF-β1. Methods: HK-2 cells induced by TGF-β1 were treated with various EM concentrations for 24 and 48 h, respectively. According to the cell proliferation changes, the best concentration of EM was selected. After HK-2 cells were stimulated with TGF-β1 for 24 h, cells were collected for Western blot and RT-PCR analysis. Protein expression of fibrosis related molecule Collagen IV was de- tected by Western blot. Protein expression of epithelial-mesenchymal transition related factors α-SMA and E-Cadherin were detected by Western blot. RT-PCR was used to detect the protein level of mesenchymal cells marker α-SMA. Results: The best concentration of EM was 40 μM by MTT analysis. Compared with TGF-β1, TGF-β1+EM had reduced expressions of Collagen IV (P<0.05). And compared with TGF-β1, TGF-β1+EM had reduced expressions of α-SMA (P<0.05). However, E-Cadherin protein expression was increased (P<0.05). RT-PCR results showed that the expression level of α-SMA mRNA in TGF-β1+EM group was lower than that in TGF-β1 group (P<0.05). Conclusion: Emodin can relieve renal tubulointerstitial fibrosis by inhibiting transdifferentiation of renal tubular epithe- lial cells. |
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