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Please use this identifier to cite or link to this item: http://ir.hwai.edu.tw:8080/ir/handle/310996100Q/1705

Title: 探討20-Hydroxyecdysone於第一型乙型轉型生長因子誘導腎近端小管細胞纖維化的角色
The role of 20-hydroxyecdysone in attenuating TGF beta-1-induced renal cellular fibrosis in proximal tubule cells
Authors: 陳韋銘
Chen, Wei-Ming
Keywords: 腎纖維化
Renal fibrosis
Epithelial-to-Mesenchymal transition (EMT)
20-hydroxyecdysterone (20-HE)
Date: 2011
Issue Date: 2013-12-20 15:51:23 (UTC+8)
Abstract: 纖維化是末期腎病變的主要特徵,造成腎小管不可逆性的損傷。先前的文獻指出,在糖尿病動物模型20-hydroxyecdysone (又稱20HE) 能夠降低腎臟損傷。然而20HE對於腎臟纖維化的調控角色仍然需要進一步的研究。方法: 細胞培養近端腎小管上皮細胞 (HK-2) 利用48小時乙型轉型生長因子TGF-1的刺激誘導作為腎細胞纖維化的體外模型,透過不同濃度的20HE (0到50M) 處理24小時,以觀察細胞內外的蛋白質的改變。利用酵素連結免疫吸附法分析細胞外的纖維蛋白表現。西方墨點法與免疫螢光染色偵測乙型轉型因子訊息路徑(TGF-1/Smad pathway) 的訊息傳遞蛋白(包含Smad2/3,4以及7),上皮-間質轉換的標記蛋白 (e.q. E-cadhirin與 -smooth muscle actin) 以及EMT的轉錄調節者Snail蛋白的表現。結果: 20HE逆轉TGF-1所造成細胞內外纖維蛋白的堆積。同時20HE能夠增加TGF-1的負向調節蛋白Smad 7的表現,降低Smad 2/3 的蛋白質表現以及磷酸化。另一方面20HE同時抑制上皮-間質的轉換過程。並且也抑制Snail 蛋白的表現。 結論:我們的研究指出20HE 在近端腎小管上皮細胞的纖維化模型中具有對抗纖維形成的潛力,更重要的是20HE能夠對抗纖維形是透過調節乙型轉型生長因子Smad訊息路徑和上皮-間質轉換過程。
Renal fibrosis is end stage of diabetes kidney diseases, which causes irreversible progressive proximal tubular injury. In the previous study, 20-hydroxyecdysterone (20HE) attenuated renal injury in diabetes models. However, the fibrosis regulatory role of 20HE remains to be investigated. Methods: The proximal tubular epithelial cell (designated as HK-2) were treated for 48 hours with TGF-1 (5 ng/ml) in different concentrations of 20HE (0 to 50 M/ml) in the last 24 hours of culture. The extracellular fibronectin was measured by ELISA assay. Western blot and immunofluorescence were used to evaluate the expression of TGF-1/Smads transducer (including Smad2/3, 4, and 7), epithelial and mesenchymal markers (e.q. E-cadhirin and α-smooth muscle actin) and snail (a transcriptional regulator for EMT). Results: 20HE reversed TGF-1-induced increase in fibronectin (both intracellular and extracellular fibronectin). Simultaneously, 20HE reversed TGF-1-induced down- regulation of Smad7. In addition, 20HE attenuated TGF-1-induced up-regulation of Smad 2/3 and its phosphorylation. 20HE was significantly restored in TGF--induced downregulation of E-cadherin and prohibited gained expression of -SMA. In addition, snail proteins were down-regulated by 20HE. Coclusion: We suggest that 20HE is a potential fibrosis antagonist for proximal tubule cells. More importance, 20HE appear to be an anti-fibrosis agent was through regulation of TGF-1/Smad pathway and EMT.
Appears in Collections:[生物科技系暨生物醫學研究所] 博碩士論文

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