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研究生:吳晉東
研究生(外文):Chin-Tung Wu
論文名稱:藥用石斛之分子鑑定及輔助調節過敏體質之研究
論文名稱(外文):The Study of Molecular Diagnosis and Immunomodulation in Herba Dendrobii
指導教授:蔡新聲蔡新聲引用關係陳宗禮陳宗禮引用關係
口試委員:林文川謝長奇郭昭麟
口試日期:2011-07-13
學位類別:博士
校院名稱:國立中興大學
系所名稱:農藝學系所
學門:農業科學學門
學類:一般農業學類
論文種類:學術論文
論文出版年:2011
畢業學年度:99
語文別:中文
論文頁數:114
中文關鍵詞:藥用石斛
外文關鍵詞:Herba Dendrobii
相關次數:
  • 被引用被引用:2
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  • 收藏至我的研究室書目清單書目收藏:1
蘭科石斛屬植物的莖是一種名貴的中草藥,因此時常有偽品出現造成混淆,因此鑑定真偽品的方法是一個必要的課題。本研究,以Internal Transcribed Spacers(ITS)的分析確定11種石斛及兩個偽品(Pholidota articulata and Flickingeria comate)的親緣關係。結果顯示,13個物種ITS總長度介於635至641 bp,其GC含量(ITS1+ 5.8 S+ ITS2)約50.55%至57.25%。石斛屬ITS序列之間約有平均13.20%的差異,而石斛與P. articulata and F. comate 則分別有42.00%和29.00%的差異。分子演化樹顯示,石斛屬遺傳距離較為接近,而偽品距離較遠並各自獨立。因此,ITS是一種可以被用於鑑別藥用石斛的分子標記。
黃花石斛為「石斛」藥材基原之一。目前已知石斛屬植物會產生多種生物活性代謝物,包括槲皮素。異位性皮膚炎(atopic dermatitis, AD)是一種慢性過敏性發炎皮膚疾病,好發於幼童。本試驗以2,4,6-trinitro-1-chrolobenzene(TNCB)重複塗抹誘導BALB/c小鼠耳後皮膚成為異位性皮膚炎之模型。這些小鼠表現之特徵,包括皮膚紅疹、肥大細胞(mast cell)數量增加與免疫球蛋白E(IgE)濃度增加及相關細胞激素改變。本研究顯示,黃花石斛之乙酸乙酯萃取物(DtE)對異位性皮膚炎模型動物具有保護作用。結果發現口服DtE處理組之IL-4及IgE濃度有顯著下降,IFN-γ濃度則顯著上升;脾臟之調節細胞(Treg cells)亦有顯著增加並減少Th2細胞分化;處理組小鼠之皮膚肥大細胞亦與對照組顯著減少。本研究顯示口服DtE能有效降低小鼠異位性皮膚炎之症狀。


Herba Dendrobii, commonly known as “Shi-hu”, has been used as a precious traditional Chinese medicine. It is expensive and adulteration are common due to high demand. The method to distinguish the herb from adulterant species is necessary. In the present study, internal transcribed spacers (ITS) region-based analysis was employed to ascertain the phylogenetic relationship among the 11 Dendrobium and two adulterant species Pholidota articulata and Flickingeria comate. Results showed that the length of the ITS regions among the thirteen species ranged from 635 to 641 bp and the GC ratio in ITS (ITS1 + 5.8S + ITS2) regions ranged from 50.55% to 57.25%. Dendrobium species was significantly different from one another by an average of 13.20% and from P. articulata and F. comate by 42.00% and 29.00% respectively. The molecular phylogenetic trees indicated that most of Dendrobium species are closely related and share common clad while both the adulterants outgroup and have separated clad. Therefore, ITS regions can be used as a molecular marker to differentiate medicinal Dendrobium spp. from one another and also from adulterants.
Dendrobium tosaense Makino is one of the most valuable Chinese medicines known as “Shi-hu” and well developed functional health food in Taiwan. Atopic dermatitis (AD) is a chronic inflammatory skin disease that occurs mainly in childhood. The pathogenesis of atopic dermatitis has been studied in BALB/c mice produced by administration of 2,4,6-trinitro-1-chrolobenzene (TNCB). These mice exhibit features of chronic contact dermatitis, including skin rash, an increased number of mast cells, and elevated serum IgE levels accompanied by a shift in cutaneous cytokine expression. In the current study, a standardized ethyl acetate extract of D. tosaense (DtE) was used to protect these mice from the TNCB-induced skin lesions simulating atopic dermatitis. Activated T cell differentiation, cytokine modulation, allergen-specific antibody estimation, skin pathology, and mast cell numbers in the skin lesions were determined. The resulting cytokine profiles and IgE level showed DtE to significantly reduce Th2 polarization. An increased number of modulated natural T regulatory cells were accompanied by increased immunosuppression in the spleen and decreased mast cell infiltration into the skin lesions. Our results indicate that the administration of DtE can modulate cytokine expression and T cell subpopulations by regulating mast cell infiltration and thus alleviate the atopic dermatitis.


中文摘要…………………………………………………………………1
英文摘要…………………………………………………………………3
第一章前言……………………………………………………………5
第二章前人研究
第一節藥用石斛之簡介………………………………………8
第二節分子標記於中草藥之應用…………………………30
第三節異位性皮膚炎及免疫系統…………………………37
第三章利用內轉錄間隔區鑑別藥用石斛之親緣關係
一、前言……………………………………………………47
二、材料與方法……………………………………………48
三、結果……………………………………………………53
四、討論……………………………………………………64
第四章黃花石斛萃取物對異位性皮膚炎之影響
一、前言……………………………………………………66
二、材料與方法……………………………………………69
三、結果……………………………………………………77
四、討論……………………………………………………88
第五章綜合討論………………………………………………………93
第六章參考文獻………………………………………………………99


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