過敏性氣喘寒熱證型動物模型之建立

ALLERGIC ASTHMA ANIMAL MODEL WITH COLD AND HOT “ZHENG”

辨證論治是我國傳統醫學診斷治療上的獨特理論，中醫將哮證急性期區分為寒哮與熱哮，為正確評估中醫藥平喘方劑的療效及研究其機轉，本實驗建立過敏性氣喘之寒、熱證動物模型。本文利用寒性（黃芩、黃連、梔子）及熱性（肉桂、乾薑、附子）兩組藥性不同的中藥餵食小白鼠，造成小白鼠體質的改變，並用免疫方法誘發氣喘，以建立有寒、熱證型之別的氣喘動物模型。實驗中以觀察實驗動物之外觀；測量其生理的變化，包括體重、體溫與心率；以家塵　抗原（Dermatophagoides pteronyssinus group 5 allergen ）腹腔注射致敏小白鼠，並由呼吸道吸入抗原誘發氣喘後，觀察呼吸道病理組織切片；以酵素連結免疫分析法（ELISA ）測定血清中特異性抗體IgG 、IgE 及氣管肺泡沖洗液中細胞間白素-4 和伽瑪干擾素的濃度變化來說明寒、熱證氣喘動物生理、病理及氣喘免疫反應的差異。 結果發現寒組、熱組及對照組小鼠外觀有顯著差異，熱組動物活動力增加，而寒組則下降；寒組動物的心率下降（700 ±34 次/min VS 600 ±103 次/min, p < 0.05 ）；三組動物之病理組織切片有明顯差異，熱組小鼠呼吸道炎症最為嚴重，寒組最輕微，對照組則介於兩者之間。血清中特異性抗體IgG 、IgE 三組顯著無差異，而氣管肺泡沖洗液中細胞間白素-4 和伽瑪干擾素以熱組較高。因此我們認為熱證型對過敏免疫發炎反應有加強的效果，本實驗為初步探討氣喘寒、熱證型動物之生理、病理及氣喘過敏免疫反應，但氣喘是相當複雜的疾病，仍須進一步研究才能了解其全貌。“Zheng” is the unique theory of diagnosis and treatment in Traditional Chinese Medicine (TCM). According to this theory, the acute phase of asthma is divided into cold “Zheng” and hot “Zheng”. The aim of this study was to establish an asthma murine model with cold “Zheng” and hot “Zheng” to evaluate the efficacy of TCM for the treatment of bronchial asthma and to study the mechanisms. First, we fed Scutellariae Coptis Gardeniae and Aconitum Cinnamomum Zingiber separately to BALB/c mice to change their nature to cold and hot. In this experiment, the animals’ appearances and changes of physical states, including body weight, body temperature, and heart rate were observed and recorded to estimate the effects of the Chinese herbal medicine. Then, we sensitized the mice using intraperitoneal injection of dust mite allergens (Dermatophagodes pteronyssinus group 5, Der p 5) and induced the allergen-specific airway hyperreactivity using inhalational challenges. Results of the pathological studies of the lung and trachea were observed to determine the severity of inflammation. The concentration of Der p 5-specific IgG and IgE antibodies in serum and cytokines IL-4 and INF-γ in bronchoalveolar larvage fluid were assessed using ELISA to clarify the differences of allergic immunoresponses between the animals with cold and hot “Zheng”. We found that the animals with cold “Zheng” and hot “Zheng” were very different in appearance. The animals with hot “Zheng” were much more active than the animals with cold “Zheng”. According to the results of pathological examinations, the animals with hot “Zheng” had the most severe airway inflammation as shown by the concentrations of both interleukin-4 and interferon-γ which were the highest among the three groups. Therefore, we suggest that the physiology and immunoresponses are quite different between the mice with cold “Zheng” and hot “Zheng”, and the production of IL-4 may play a role in the differences.