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研究生: 余宛真
Weng-Jean Yu
論文名稱: 內毒素處理下黃芩素對誘導型一氧化氮合成酵素與第二型環氧化酵素的影響
The effect of baicalin on inducible nitric oxide synthase and cyclooxygenase-2 expression in lipopolysaccharide-treated macrophage RAW264.7
指導教授: 李銘亮
Li, Ming-Liang
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2001
畢業學年度: 89
語文別: 中文
論文頁數: 66
中文關鍵詞: 黃芩素一氧化氮前列腺素E2誘導型一氧化氮合成酵素第二型環氧化酵素腫瘤壞死因子-(alpha)細胞株RAW264.7
英文關鍵詞: baicalin, nitric oxide, prostaglandin E2, inducible nitric oxide synthase, cyclooxygenase-2, TNF-(alpha), RAW264.7 cell line
論文種類: 學術論文
相關次數: 點閱:272下載:1
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  • 中文摘要
    由中藥黃芩根部萃取出來的黃芩素,在近來研究上具有抗氧化和抗發炎的藥效。許多研究指出,體內過量一氧化氮(nitric oxide, 簡稱NO)、前列腺素E2(prostaglandin PGE2, 簡稱PGE2)、自由基和腫瘤壞死因子-a(tumor necrosis factor-a, 簡稱TNF-a)與敗血性休克的形成有關。而過量NO與PGE2的生成分別是經由誘導型一氧化氮合成酵素(inducible nitric oxide synthase, 簡稱iNOS)與第二型環氧化酵素(cyclooxygenase-2, 簡稱COX-2)所產生。本研究是以內毒素(lipopolysaccharide, 簡稱LPS)刺激細胞株RAW264.7探討黃芩素對於NO、PGE2和TNF-a生成量的影響與iNOS及COX-2的表現。
    從我們的結果可知,在以適當濃度LPS(仍可保持良好的細胞活性)刺激下的細胞株RAW264.7,隨著黃芩素處理濃度(10~50 mM)的增高,抑制NO、PGE2和TNF-a生成的效果越好。再者,黃芩素也可以抑制因LPS誘導所產生的iNOS及COX-2的mRNA與蛋白質表現。由結果可推知,黃芩素抑制iNOS及COX-2的mRNA與蛋白質表現的機轉可能是經由抑制TNF-a過量的生成。因此,黃芩素對於治療敗血性休克有很大潛力。另外,以50 mM黃芩素處理細胞株RAW264.7可降低細胞週期中S期和G2/M期的比例,所以黃芩素可能也可當為治療腫瘤的藥劑。

    Abstract
    It has been demonstrated that baicalin extracted from the root of Huangquin, a Chinese herb medicine has potential antioxidant and anti-inflammatory effect. Many studies have indicated that overproduction of nitric oxide (NO), from inducible nitric oxide synthase (iNOS), prostaglandin E2 (PGE2) from cyclooxygenase-2 (COX-2), free radical, and tumor necrosis factor-a (TNF-a) is involved in the pathogenesis of septic shock. In this study, we evaluated the effect of baicalin on these mediators and iNOS as well as COX-2 expression in lipopolysaccharide (LPS)-treated RAW264.7 macrophage.
    Our results demonstrated that baicalin ( 10~50 mM ) concentration-dependently inhibited LPS-induced NO、TNF-a and PGE2 production in LPS-treated RAW264.7 macrophage without an appreciable cytotoxic effect. Furthermore, baicalin inhibited the expression of LPS-induced iNOS and COX-2 proteins and mRNAs. These results indicated that the inhibition of TNF-a production by baicalin maybe contribute to the suppression of iNOS and COX-2 expressions. Thus, baicalin may be beneficial for treatment of endotoxaemia. Besides, treatment of the RAW264.7 macrophage with 50 mM baicalin decreased the S phase and G2/M phase ratio about 15% in the cell cycle. Thus, baicalin may be also a potential therapeutic drug for tumor therapy.

    目錄 目錄………………………………………………………………….…Ⅰ 圖目錄…………………………………………………………………..Ⅱ 中文摘要…………………………………………………………………1 Abstract..……………………………………………………………...…..2 前言……………………………………………………………………....3 實驗設計…………………………………………………………………9 材料與方法…………………………………………………………..…13 實驗結果…………………………………………………………….….20 討論………………………………………………………………….….25 結論………………………………………………………………….….30 參考文獻…………………………………………………………….….31 附錄一…………………………………………………………………..65 附錄二…………………………………………………………………..66 圖目錄 圖一、不同濃度內毒素對一氧化氮生成量的影響。………………..…42 圖二、不同濃度內毒素對細胞活性的影響。…………..……………....43 圖三、內毒素刺激下,一氧化氮的生成與時間的關係。……………....44 圖四、內毒素刺激下,不同濃度黃芩素對一氧化氮生成量的影響。…45 圖五、內毒素刺激下,不同濃度黃芩素對細胞活性的影響。…………46 圖六、內毒素刺激下,不同濃度黃芩素對TNF-α生成量的影響。…...47 圖七、內毒素刺激下,不同濃度黃芩素對PGE2生成量的影響。……..48 圖八、內毒素刺激下,一氧化氮合成酵素(iNOS)mRNA表現與時間的關係。………………………………………………………………49 圖九、內毒素刺激下,黃芩素對一氧化氮合成酵素(iNOS)mRNA表現的影響。……………………………………………………………50 圖十、內毒素刺激下,黃芩素對一氧化氮合成酵素(iNOS)蛋白表現的影響。………………………………………………………………51 圖十一、內毒素刺激下,第二型環氧化酵素(COX-2)mRNA表現與時間的關係。…………………………………………………………52 圖十二、內毒素刺激下,黃芩素對第二型環氧化酵素(COX-2)mRNA表現的影響。……………………………………………………………53 圖十三、內毒素刺激下,黃芩素對第二型環氧化酵素(COX-2)蛋白表現的影響。…………………………………………………………54 圖十四、不同濃度內毒素對細胞內DNA的變化。……………………55 圖十五、不同濃度黃芩素對細胞內DNA的變化。………………...….58 圖十六、經同步化處理後的細胞株,處理不同濃度黃芩素對細胞內DNA的變化。………………………………………………………...…62

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