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研究生: 謝鳴綺
Hsieh, Ming-Chi
論文名稱: 黃芩萃取物及其組成分抑制痤瘡丙酸桿菌誘導之發炎反應
Radix Scutellariae extracts and its components suppress Propionibacterium acnes-induced inflammation
指導教授: 吳文惠
Wu, Wen-Huey
學位類別: 碩士
Master
系所名稱: 人類發展與家庭學系
Department of Human Development and Family Studies
論文出版年: 2015
畢業學年度: 103
語文別: 中文
論文頁數: 76
中文關鍵詞: 痤瘡黃芩抗發炎介白素-1β介白素-8
英文關鍵詞: Acne vulgaris, Radix Scutellariae, wogonin
論文種類: 學術論文
相關次數: 點閱:121下載:6
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  • 痤瘡桿菌(Propionibacterium acnes)是痤瘡等發炎疾病的重要因子,會刺激單核球等細胞分泌大量促發炎細胞激素IL-1β、IL-8,造成發炎反應。黃芩是一種傳統中草藥,在中醫合併其他藥物用來治療痤瘡,其主要活性成分有Baicalin、baicalein、wogonin和oroxylin A,具有抗發炎、抑制腫瘤生成等效果。
    本研究以痤瘡桿菌刺激人類單核球細胞(THP-1)為發炎模式,先測試黃芩正丁醇、乙酸乙酯(EtOAc)粗萃物的抗發炎效果,發現黃芩EtOAc粗萃物及其所含有的七種黃酮類化合物oroxylin A、wogonin、7-O-methylwogonin、skullcapflavone II、5,7, 4'-trihydroxy-8-methoxyflavone、5,6,2'-trihydroxy-7,8-dimethoxyflavone
    和ganhuangenin,均可顯著抑制IL-1β和IL-8分泌。其中wogonin效果最佳,抑制IL-1β和IL-8分泌之IC50分別為4.9μM、8.7μM,其次為5,7,4'-trihydroxy-8-methoxyflavone抑制IL-1β和IL-8分泌之IC50為11.3μM和10.2μM。之後以小鼠耳朵注射痤瘡桿菌模式,檢測耳朵厚度和組織均質液IL-1β、IL-6和TNF-α濃度,發現七個化合物皆可抑制耳朵發炎和腫脹現象且抑制三種細胞激素的分泌。
    綜合而論,黃芩萃取物和其活性成分能有效抑制痤瘡桿菌引起的發炎反應,具改善痤瘡潛力

    Propionibacterium acnes (P. acnes), the main pathogenic bacteria of acne vulgaris, stimulates IL-1β and IL-8 secretion in immunocytes. Radix Scutellariae (SB) is a common Chinese medicinal herb in Asia, having been used to treat acne. Our research evaluated the effects of SB extracts and its components on inflammation in THP-1 cells stimulated by P. acnes. The results showed that the ethylacetate (EtOAc)
    extract of SB significantly inhibited P. acnes induced IL-1β and IL-8 secretion. Seven compounds, oroxylin A, wogonin, 7-O-methylwogonin, skullcapflavone II, 5,7, 4'-trihydroxy-8-methoxyflavone, 5,6,2'-trihydroxy-7,8-dimethoxyflavone, and
    ganhuangenin, isolated from SB EtOAc extract inhibited IL-1β and IL-8 secretion. Among them, wogonin had the best effect with IC50 of 4.9μM and 8.7μM for IL-1β and IL-8, respectively. Next was 5,7,4'-trihydroxy-8-methoxyflavone with IC50 of 11.3μM and 10.2μM for IL-1β and IL-8, respectively. Subcutaneous injection of P. acnes into mouse ears increased ear thickness, and levels of IL-1β, IL-6 and TNF-α in ear homogenates. Each of the 7 compounds simultaneously injected with P. acnes reduced the ear thickness and the production of these 3 cytokines. In summary, EtOAc extract of SB and its seven components significantly inhibit P. acnes induced inflammation, and may improve acne vulgaris.

    中文摘要 iii 目錄 v 圖目錄 vii 第一章 緒論 1 第一節 研究動機 1 第二節 研究目的 2 第二章 文獻探討 3 第一節 痤瘡桿菌與發炎 3 壹、 痤瘡與痤瘡桿菌 3 貳、 痤瘡桿菌和發炎反應 6 參、相關細胞激素 8 第二節 痤瘡的治療 10 第三節 實驗材料 12 壹、 黃芩(Huangqin; Radix Scutellariae ) 12 第三章 材料與方法 16 第一節 實驗架構 16 第二節 研究材料 17 壹、 藥品與試劑 17 貳、 儀器設備 21 第三節 實驗方法 22 壹、 樣品製備 22 貳、 細胞培養 22 參、 痤瘡桿菌 (Propionibacterium acnes, BCRC10723) 培養 24 肆、 細胞存活率測定-MTT assay 25 伍、 in vitro抑制IL-1β、IL-8生成之活性評估-ELISA assay 26 陸、 in vivo抗發炎活性評估 29 柒、統計分析 34 第四章 結果 35 第一節 in vitro抗發炎作用 35 壹、 細胞存活率測定(MTT) 35 貳、 抑制IL-1β、IL-8生成之活性評估(ELISA assay) 38 第二節 in vivo抗發炎活性評估 52 第五章 討論 62 第一節 黃芩粗萃物in vitro 抗發炎活性評估 62 第二節 FL1-FL7的in vitro和in vivo 抗發炎活性評估 63 第三節 結論 66 第六章 參考文獻 67

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