治療過敏性氣喘中藥複方之研究;
A Study of Traditional Chinese Medicine Formulae for Anti-asthma Treatment

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题名:	治療過敏性氣喘中藥複方之研究;A Study of Traditional Chinese Medicine Formulae for Anti-asthma Treatment
作者:	張東迪;Tung-Ti Chang
贡献者:	中國醫藥大學：中西醫結合研究所
关键词:	動物試驗;臨床試驗;過敏性氣喘;中藥複方;嗜伊紅性白血球顆粒蛋白;綠色螢光蛋白;STA-1;mMMDT;Traditional Chinese Medicine;Allergic asthma;Eosinophil cationic protein;Green fluorescent protein
日期:	2006-06-05
上传时间:	2009-08-05 15:00:47 (UTC+8)
摘要:	許多研究顯示全球過敏性疾病，不僅盛行率一直上升，以氣喘為例，疾病發作的嚴重度及死亡率也不斷上升。氣喘患者經常使用類固醇控制，然而長期使用類固醇卻有許多不良副作用而為人們所畏懼。因此在亞洲國家已經應用千百年之中醫藥便吸引西方國家嘗試用來治療氣喘。慢性呼吸道發炎，向來是西方醫學對氣喘之主要定義，本研究從塵蟎造型之過敏性氣喘動物實驗開始，評估中藥複方STA-1對氣喘之抗發炎效果。結果發現STA-1對致敏小白鼠之呼吸道抗發炎之效應，顯示能明顯降低小白鼠D p 5-specific IgE之合成，明顯減低嗜中性白血球與嗜伊紅性白血球之呼吸道浸潤，以及降低過敏原引起之呼吸道過度反應性。我們以此方設計一雙盲安慰劑對照控制臨床試驗，並以現代醫學評估氣喘之指標為終末測量(endpoint outcome measure)，包括肺功能、臨床症狀、血中總IgE、Der p 5-specific IgE濃度以及類固醇用量。結果顯示，STA-1能改善輕中度(mild-to-moderate)氣喘患者之臨床症狀，增進患者肺功能，降低其類固醇使用量，並抑制患者血清總IgE與Der p 5-specific IgE之合成。我們進一步以含五味中藥，以更簡單之麥門冬湯加減方(mMMDT)設計一雙盲安慰劑對照控制臨床試驗，結果顯示治療組FEV1値呈現明顯上升，大多數受試者之肺功能明顯改善。更重要的是，在症狀如咳嗽、喘鳴與呼吸困難之嚴重度上皆獲得緩解。治療組血中總IgE顯示下降之趨勢。除此，應用此方劑治療氣喘尚屬安全。進一步為了探討中藥對分子與基因之調控機制，我們選擇與過敏關係相當密切之嗜伊紅性白血球顆粒蛋白(ECP)做研究，因為ECP通常被認為代表嗜伊紅性白血球被活化之指標之一。因此我們以人類白血病之細胞株(HL-60 Clone 15)模擬嗜伊紅性白血球，並以即時反轉錄多鏈酶反應(real time RT-PCR)做為監測基因表現之方法。結果顯示本法可作為探討ECP表現之細胞模型。最後我們希望能將ECP之驅動子(promoter)基因接合一含產生綠色螢光蛋白(green fluorescent protein)之報導基因(reporter)，將之轉殖到HL-60 Clone 15以建立一新的穩定之轉基因細胞，期望建立快速篩選與探討包括本研究相關之中藥免疫機制之體外模型，以之做為嗜伊紅性白血球顆粒蛋白表現之生物感應器(biosensor)。結果我們成功選殖出ECP驅動子基因並將之插入綠色螢光載體(vector)之多選殖位址(multiple cloning site)，並成功構築一新的含ECP驅動子之載體，綠色螢光基因即緊臨於ECP驅動子下位(down-stream)。 The increasing incidence and prevalence of asthma in the world continue to make it a global health concern. The morbidity and mortality of asthma have increased over the last two decades. The most common treatment nowadays is the use of anti- inflammatory drugs, including steroids. However, long-term steroid therapy is often associated with multiple sidedebilitating effects. On the other hand, Traditional Chinese medicine (TCM), used in Asia for centuries, has thus become an attractive interest as a source of therapies for asthma. We designed an in vivo study in STA-1, a TCM formula on allergen-induced bronchial inflammation and hyperreactivity in mite-sensitized mice. We have demonstrated that STA-1 could, 1) reduce the synthesis of Der p 5-specific IgE tremendously, 2) significantly down-regulate the influx of neutrophils and Eosinophil into airway, and 3) decrease allergen-induced specific airway hnyperreactivity. Our study by double-blinded placebo control trial showed that STA-1could provide improvement in lung function and relieve asthma symptoms without adverse effects. In addition, the decreasing of the serum total IgE and the steroid-sparing effect suggests its role in modulating immune response. Furthermore, we design another clinical study on a simplified formula mMMDT in allergic asthma. The results showed that the mMMDT provided improvements in lung function and relieved asthma symptom. Since its efficacy and safety, we consider mMMDT as a credible treatment regimen for mild to moderate persistent asthma. Serum levels of ECP may reflect the degree of bronchial inflammation in patients with asthma. We set up a cell model by using real-time RT-PCR for analyzing the effect of STA-1 on ECP expression. We also successed to construct a plasmid containing an fusion of two genes, GFP and ECP promoter for monitoring indirect quantitative correlation between the fluorescence intensity of GFP and the regulatory activity of the upstream ECP promoter. The final aim of this study is to investigate the mechanism of TCMs on regulation of ECP in this cell model.
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