Study on the Antioxidant Activity of Aqueous Extracts from Traditional Chinese Herbs in CHO-K1 and ECV304 Cells

• Main Authors: Huang Ching, 黃苑清
• Other Authors: Tsung-Shing Wang
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/90743900981415164500

碩士 === 中山醫學大學 === 生物醫學科學學系碩士班 === 97 === BACKGROUND: A variety of evidence suggests that chronic oxidative stress may accelerate aging and increase risk of carcinogenesis. Therefore, exploring natural compounds that possess high antioxidant activity should be useful in both treating cancer and delaying aging. OBJECT: The object of this study was to evaluate the anticlastogenic effects of aqueous extracts from five traditional Chinese medicines with high free radical scavenging activity on two different cell models (CHO-K1 and ECV304 cells). METHODS: The comet assay was used for DNA damage detection, crystal violet stain was used for cell proliferation detection, Western blotting was used to detect the protein level of antioxidant enzymes (HO-1, catalase, SOD-1) and DNA repair proteins (Ogg1, MTH-1, and MYH). RESULTS: Pre-treatment of aqueous extracts derived from five different Chinese medicinal herbs (Dioscorea alata山藥, Ligusticum chuanxiong 川芎, Rehmania glutinosa 生地黃, Paeonia suffruticosa 牡丹皮, and Artemisia scoparia 茵陳蒿) for 48-hr decreased the DNA strand breaks level induced by H2O2 in CHO-k1 and ECV304 cells as compared with untreated control cells. The percent inhibition of H2O2-induced DNA strand breaks was found to be in the range 18-63%. Co-treatment of aqueous extracts derived from five different Chinese medicinal herbs, only D. alata and L. chuanxiongcould significantly inhibit the CuSO4-induced DNA strand breaks in CHO-K1 and ECV304 cells. However, the inhibition of CuSO4-induced DNA strand breaks by R. glutinosa and Artemisia scoparia 茵陳蒿was found only in ECV304 cells but not CHO-K1 cells. The percent inhibition of CuSO4-induced DNA strand breaks by extracts of TCM was found to be in the range 15-38%. In ECV304 cells, forty-eight hours pre-treatment of five different traditional Chinese medicines could significantly up-regulate Bax, Bcl2, MUTYH, MTH1 or HO-1 protein expression The level of L. chuanxiong -induced HO-1 expression was attenuated by Wortmannin (a phosphatidylinositol 3-kinase, PI3K inhibitor), SR11302 (a AP-1 inhibitor), and Bay 11-7082 (a NFκB inhibitor). CONCLUSION: These results suggest that the anticlastogenic activities of aqueous extracts of L. chuanxiong, A. scoparia, R. glutinosa, and P. suffruticosa may be partially through the up-regulation of antioxidant and DNA repair enzymes expression and copper-chelating activity. The data also suggest that L. chuanxiong- induced HO-1 expression, at least in part, through PI3K, AP-1 and NFκB signaling pathways.