PHSeaFood:香茹草乙醇萃取物抗發炎機制之探討
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From PHSea
Revision as of 01:40, 1 April 2006
資料來源：全國博碩士論文查詢系統
- 系統編號:091CNUPS111006
- 出版年:2003
- 研究生:王麗淑
- 研究生(英文姓名):Lisu Wang
- 論文名稱：香茹草乙醇萃取物抗發炎機制之探討
- 英文論文名稱:Anti-inflammatory Mechanism of Ethanol Extract of Hsiang Ju
- 指導教授:吳明娟 博士
- 指導教授(英文姓名):Dr. Ming-Jiuan Wu
- 學位類別:碩士
- 校院名稱:嘉南藥理科技大學
- 系所名稱:生物科技研究所
- 學號:G9015013
- 學年度:91
- 語文別:中文
- 論文頁數:112
- 關鍵詞:香茹;巨噬細胞;誘導型一氧化氮;誘導型環氧化&; 細胞激素
- 英文關鍵詞:Glossogyne tenuifolia;nitric oxide (NO);inducible nitric oxide synthase(iNOS); Cytokine;NF-kappa B;luteolin-7-glucoside;oleanolic acid
- 被引用次數:1
- [ 摘要 ]
香茹(Glossogyne tenuifolia, GT)是澎湖島的原生植物，民間將其作為退熱、解毒、抗發炎之用藥，但目前並無與抗發炎作用機轉及有效成分相關的文獻報導。
當巨噬細胞受到內毒素脂多醣LPS（lipopolysaccharide）或免疫刺激（Interferon-gamma, IFN-）時，會產生各種發炎調節因子，誘發一系列的發炎反應。本研究主要以LPS刺激誘導的老鼠巨噬細胞株RAW264.7做為反應模式，探討香茹的乙醇萃取物(GT)對於促發炎的調節因子的釋放及相關調控基因的影響。
此論文第一部分主要探討GT對LPS刺激誘導的老鼠巨噬細胞株RAW264.7的亞硝酸鹽產生的抑制的效果。結果顯示GT (0.05-0.25 mg/ml)對一氧化氮產生的抑制作用，隨濃度的增加遞增。利用西方吸漬法及RT-PCR分析誘導性一氧化氮合成酶(iNOS)的基因表達時發現，GT對iNOS 基因表達的抑制，亦隨濃度的增加遞增。另外利用添加actinomycin D 抑制RNA生合成時發現，GT並不會影響iNOS mRNA的穩定度。因此GT主要是藉調控iNOS的轉錄來抑制一氧化氮合成酶的表達，進而抑制一氧化氮的生合成。此外，GT與nuclear factor-B(NF-B)活化的抑制劑pyrrolidine dithiocarbonate (PDTC) 或NOS的抑制劑aminoguanidine(AG)共同作用在活化的RAW264.7時，可協同抑制一氧化氮的產生。
在此論文的第二部分主要探討香茹草的乙醇萃取物(GT)對於花生四烯酸的代謝終產物(PGE2)的抑制。由結果中發現，雖然GT能夠抑制PGE2的產生，但對於COX-2 mRNA及蛋白質的表達皆無明顯抑制的效果，因此推論GT可能抑制了COX-1途徑或是影響了COX酵素的活性或是影響了PGE2合成酵素的生合成等。
論文的第三部分則是探討GT對一級發炎調節因子的釋放的影響。在實驗結果中顯示GT能夠顯著的抑制受活化的巨噬細胞產生TNF-, IL-1 IL-6, IL-12等促發炎細胞激素。抑制效果隨GT劑量的增加而遞增。
論文的第四部分是探討GT對發炎基因共同的轉錄因子NF-B的活化的影響。利用西方吸漬法發現GT能夠顯著的抑制細胞質裡的IB的磷酸化及進一步的水解。利用轉染含NF-B responsive element及luciferase reporter gene的質體進入RAW264.7時發現，GT可顯著抑制LPS誘發的NF-B的活化。
在論文的最後一部分進一步探討香茹主要成份oleanolic acid及luteolin-7-glucoside，對nitrite、細胞激素及NF-B活化的抑制效果。我們發現此這些物質皆能抑制受LPS活化的巨噬細胞的NO, TNF-, IL-6, IL-12的產生及NF-B 的活化。然而此三種成份與PDTC及aminoguanidine無協同抑制NO的產生。
因此，本研究的主要貢獻為確立香茹草乙醇萃取物之抗發炎分子藥理機轉，並確定luteolin-7-glycoside及 oleanolic acid為其抗發炎之指標成份。
