促進史旺氏細胞增生的生藥篩選平台之建立

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Title:	促進史旺氏細胞增生的生藥篩選平台之建立 The screening platform of Traditional Chinese Medicine on Schwann cell proliferation
Authors:	薛明芳;Ming-Fang Hsueh
Contributors:	中醫學院中醫學系碩士班
Keywords:	中醫藥;周邊神經再生;纖維母細胞生長因子;Traditional Chinese Medicine;Peripheral nerve regeneration;Fibroblast-Growth-Factor-2;Foshou
Date:	2010
Issue Date:	2010-09-29 12:00:54 (UTC+8)
Abstract:	周邊神經再生是一個多因素共同影響的過程，中藥單藥及複方在促進神經再生上的研究廣泛地受到討論。本研究運用含纖維母細胞生長因子(Fibroblast Growth Factor-2)啟動子的螢光酵素酶質體轉殖入RSC 96史旺氏細胞，藉由偵測螢光酵素酶活性以尋找可能具有促進史旺氏細胞增生的中藥。依照螢光酵素酶偵測值，歸納出有理氣藥、去濕藥、清熱藥、活血化瘀藥、平肝息風藥。進一步選取中藥佛手刺激史旺氏細胞後，由西方氏墨點法偵測細胞核增生抗原和細胞週期調節蛋白表現量得知佛手可能有促進神經細胞增生的效用。以Boyden chambers觀察史旺氏細胞在加入佛手刺激後，細胞移動的情形以佛手濃度3.4μg/ml時最為顯著。以西方氏墨點法偵測FGF-2-UPA-MMPs途徑因子蛋白質表現量，發現佛手能透過FGF-2-UPA-MMPs訊息傳遞路徑增加史旺氏神經細胞移動作用。本篇論文結合中醫理論、中藥資源和採用現代醫學作定性、定量分析，探索中藥促進神經再生可能的機制，可提供將來發展促進神經再生中藥的新契機。 Numerous factors and elements are involved in the regeneration of peripheral nerve. Chinese herbs and medicine decoctions applied on nerve regeneration have been widely discussed. In this study, we transfected the luciferase plasmid with Fibroblast-Growth-Factor-2 promoter sequence into RSC96 Schwann cells. By detecting the activity of luciferase system,we screened some Chinese herbs potentially helpful to the regeneration of peripheral nerve. Further,we investigated the effect of Foshou on proliferation and migration of Schwann cells. After treating with different concentrations of Foshou,we found that 3.4 μg/ml of Foshou could induce significant proliferation of Schwann cells by MTT assay and Western blotting assay. In addition,we investigated Schwann cells migration treated with different concentrations of Foshou through the migration assay and Western blotting assay. It was found that 3.4 μg/ml of Foshou also induced marked migration of Schwann cells. Foshou enabled to stimulate FGF-2-UPA-MMPs pathway regulator and cell cycle controlling protein expression. This work demonstrates that modern molecular biology method can be combined to Traditional Chinese Medicine theory to find some new drugs promoting peripheral nerve regeneration.
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