全球過敏性疾病的盛行率愈來愈高，疾病發作的嚴重度及死亡率也不斷上升。傳統醫學治療過敏性疾病己使用數千年，目前也有愈來愈多的病患尋求中醫藥之輔助，由於中醫之“辨證”往往缺乏客觀的指標，因此“辨證”限制了中醫藥的普及和過敏性疾病新藥之開發。 本研究嘗試從臨床上典型對塵蟎過敏之過敏病人身上，依中醫傳統望、聞、問、切之臨床觀察將病人歸類，希望找出過敏性病人中醫辨證的切入點，並尋找較客觀之現代免疫學指標加以量化，結果發現用八綱中之寒、熱辨證可以將過敏性疾病的病人分為二個次族群，並發現血清嗜伊紅性陽離子蛋白與偏熱證型之病人有明顯之正相關。為了進一步研究過敏性疾病寒熱證型之機轉，我們以Der p 5致敏之氣喘小鼠為模型，使用寒熱屬性不同的中藥來造證，結果發現寒證之氣喘小鼠不但毛色、活動力減低，血清中之Der p 5特異性抗體也與熱證小鼠有所差異，支氣管肺泡沖洗液中之細胞激素在寒熱證之小鼠中也不相同，另外，從小鼠支氣管及肺葉之切片也可發現寒證的發炎程度遠低於熱證小鼠，顯示在過敏性疾病中寒證與熱證的不同屬性可能會有不同的免疫反應。進一步，我們以人類白血病(HL-60)之細胞株，使用0.5 mM之丁酸使其分化為嗜伊紅性白血球，並利用免疫染色的方法，測量寒、熱中藥對嗜伊紅性白血球釋放嗜伊紅性陽離子蛋白的影響，結果發現0.1 %的濃度下，寒藥可以抑制嗜伊紅性白血球產生嗜伊紅性陽離子蛋白，而熱藥則不能。最後，我們利用熱證評分將氣喘病人分為熱證明顯、微熱證型及非熱證型三組，抽取病人之周邊血，利用微陣列生物晶片的技術，選定96個與氣喘致病機轉相關的基因來分析，目的在研究氣喘病人之辨證分型與基因表達之差異，結果發現在不同程度熱證評分中，Th1/Th2基因表達有所不同，在熱證明顯時，IL-4、IL-13及 RANTES等基因表達上升，而GBP-1基因表達下降；反之，在非熱證型中可看到GBP-1基因表達上升而IL-4、IL-13及RANTES基因表達下降。 我們的研究顯示，在過敏性疾病的病人中，的確存在有寒、熱兩種不同屬性的病人，在熱證明顯時，不但血清中ECP上升，IL-4、IL-13及RANTES基因的表達也上升；非熱證型的病人則完全不同。是否將來可以用不同基因表達之差異來代表不同證型之過敏病人，值得進一步深入研究，或許將來可以利用基因表達之差異，發展出一個廻歸方程式及中醫辨證之診斷基因晶片，用來預測過敏病人中醫寒熱之程度，做為臨床治療及中藥新藥開發時之參考。; Abstract The prevalence and severity of allergic disease had been increasing in recent decades. There are many patients asking for traditional Chinese medicine (TCM) due to side effects of steroids. However, TCM is difficult to prove in clinical trials although it used to treat asthma since thousand years ago. The main reason is that we can not treat a patient by TCM without patterns identification (Bien-Zheng). The definition of “Zheng” is not very clear and lack of scientific standards. This limited the popularity and development of TCM polypharmacy. In this study, we observed the allergic patients according to the principle of “observation- inspection, listening- inquiry and pulsation”. We tried to quantitate and score the “heat-Zheng” allergic patients. Our results showed that serum ECP level of “heat-Zheng” patients were significantly higher than “not heat-Zheng” patients. In order to further understand the mechanism between “heat-Zheng” and “not heat-Zheng” allergic patients, we set up the cold-Zheng and heat-Zheng mice fed with different kinds of TCM. We observed the behavior, general appearance and changes of physical states of BALB/c mice. Then, we sensitized the mice with Der p 5 and induced the allergen-specific airway hyperreactivity using the method of inhalation challenge. Our results showed that there were quite different in general appearance and behavior between cold Zheng and heat Zheng mice. The mice fed with heat drugs were more active than those fed with cold drugs. The concentration of serum Der p 5 specific IgG and IgE were different in two kinds of mice. The cytokines (IL-4, INF-g) of bronchoalveolar larvage fluid were also significantly different. Besides, the “heat Zheng” mice had more severe airway inflammation by pathology. Furthermore, we used different kinds of TCM including heat and cold drugs, cultured with HL-60 induced eosinophil cell line. The results showed that ECP positive cells were inhibited by cold drug under the concentration of 0.1%. However, heat drug did not reveal the inhibition effect at the same concentration. At last, we observed 96 asthma related genes from clinical patients. We set up a heat-Zheng score to quantitate the severity of the patient had. We used the principle of hybridization by microarray biochip to compared the relationship between different patients. The results revealed that the gene expressions of IL-4, IL-13 and RANTES were up regulated in obvious “heat-Zheng” patients but not in “not-heat-Zheng” patients. There was also a negative correlation between guanylate binding protein 1 and “heat-Zheng” scoring (P=0.072). Therefore, we suggest that there are two different kinds of “Zheng” between allergic patients from in vivo and in vitro studies.
