葛根與黨參對周邊神經再生之影響

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題名:	葛根與黨參對周邊神經再生之影響 Effect of Pueraria lobata and Codonopsis pilosula on regeneration of peripheral nerve
作者:	陳顯東;Hsien-Tung Chen
貢獻者:	中醫學院中醫學系博士班
關鍵詞:	葛根;周邊神經再生;PC12細胞;矽膠管;血清代謝物;黨參;增生;轉移;許旺細胞;Pueraria lobata;peripheral nerve regeneration;PC12 cell;silicone rubber;serum metabolite;Codonopsis Pilosula;Proliferate and migrate effect;Schwann cells
日期:	2010
上傳時間:	2010-09-29 12:00:41 (UTC+8)
摘要:	中草藥使用於臨床上已有幾千年歷史。然而，有關中草藥對周邊神經再生的影響，至今仍極少被深入研究。本實驗目的為針對葛根與黨參，利用細胞與動物模式，探討對周邊神經再生之影響。有關葛根的研究，我們利用細胞與動物實驗評估葛根血清代謝物對周邊神經再生之影響。結果顯示葛根血清代謝物在不同濃度顯著引起神經生長因子所協調的PC12細胞之神經軸突生長及synapsin Ι蛋白表現。動物實驗顯示充填有葛根血清代謝物之矽膠管做為大鼠坐骨神經端傷間距之架橋，相較於控制組，有較多成長之再生神經、較大的神經切面面積與神經內膜面積之總和、以及較多的髓鞘化軸突數目。有關黨參對周邊神經再生的影響，我們評估黨參對於史旺細胞移動與增生之影響。黨參劑量依賴性的促進增殖細胞核抗原表現、細胞存活與傷口癒合。此外，我們也發現黨參能刺激IGFs-IGFIR-Akt-Bcl2訊息路徑蛋白、細胞週期蛋白與MAPK 訊息路徑調節蛋白 ERK、 p38 之表現。在移動方面則是增加FGF-2-uPA-MMPs 路徑蛋白表現。葛根可能是一極具潛力的促進神經生長劑；黨參是極具潛力神經再生促進劑。 With a history of several thousand years, the pharmacology and clinical application of traditional Chinese medicine has been well documented. However, the effects of Chinese herbal medicine on peripheral nerve regeneration are little known. Therefore, this study investigated the effect of Pueraria lobata and Codonopsis Pilosula on regeneration of peripheral nerve. In Pueraria lobata part, we used that in vitro and in vivo evaluation of rat serum metabolites of the Pueraria lobata (SMP) on peripheral nerve regeneration. In the in vitro study, we found that the SMP (1, 0.1, 0.01 unit of the SMP) caused a marked enhancement of the nerve growth factor (NGF)-mediated neurite outgrowth and the expression of synapsin I from PC12 cells. In the in vivo study, silicone rubber chambers filled with the SMP were used to bridge a 10-mm sciatic nerve defect in rats. At the conclusion of 8 weeks, animals from the groups treated with the SMP had a relatively more mature structure with larger mean values of myelinated axon number, endoneurial area, and total nerve area when compared with those in the controls receiving the saline only. On the other hand, this study also evaluates the proliferative and migrative effects of Codonopsis Pilosula on RSC96, Schwann cells. We investigated the molecular signaling pathways, which include: (1) survival signaling, IGFs-IGFIR-Akt-Bcl2 and proliferative signaling, Cell cycle factors and MAPK pathways. (2) migrate and anti-scar signaling, FGF-2-uPA-MMPs. After treated with different concentrations (20 μg/ml，40 μg/ml，60 μg/ml，80 μg/ml，100 μg/ml) of dangshen. We observed a dose dependent proliferative effect using PCNA western blotting assay, MTT assay and the wound healing test. We also found that dangshen stimulates the protein expression of IGF-I pathway regulators, cell cycle controlling proteins and excites the MAPK signaling pathway regulators ERK and P38. Codonopsis Pilosula even stimulates the FGF-2-uPA-MMP 9 migration pathway of RSC 96 Schwann cells. Using MAPK chemical inhibitors, U0126, SB203580, and SP600125, the proliferative effects of Codonopsis Pilosula on RSC 96 cells were identified to be ERK- and P38- dependent. Based on these results we suggest that the serum metabolites of Pueraria lobata can be a potential nerve growth-promoting factor. Moreover, applying an appropriate dose of Codonopsis Pilosula with biomedical materials would be a potential approach for enhancing neuron regeneration.
顯示於類別:	[中醫學系暨碩博班] 博碩士論文

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