苓甘五味薑辛湯對反覆性塵蟎剌激氣喘小鼠之免疫調節探討

The Immunomodulatrory Effect of Ling-Gang-Wu-Weeg- Jiang-Xin-Tang on Repetitive Dermatogoides Pteronyssinus Challenged Asthmatic Mice Model

過敏性氣喘（allergic asthma）是一種慢性氣管發炎疾病，肇因於Th2淋巴細胞反應失調，造成嗜酸性白血球浸潤、肥大細胞活化、上皮細胞肥大、腺細胞活化與增生，並增加氣管過度反應及黏液的增加。歐洲塵蟎（dermatopliagoides pteronyssinus, Der p）是目前台灣造成氣喘最主要的原因之一。我們利用隔週連續5次氣管內給予Derp誘發BABL/c小鼠的慢性氣喘動物模型，來探討苓甘五味薑辛湯在治療氣喘方面的機轉。在此次實驗中我們發現苓甘五味薑辛湯可以明顯抑制Der p激發小鼠肺泡沖洗液的發炎細胞總數、單核球、嗜中性白血球和部份嗜酸性白血球。在肺泡沖洗液中，苓甘五味薑辛湯明顯抑制TGF-p和TNF-a濃度，且增高了Thl細胞激素IL-12的濃度。此外，肺部組織EMSA和免疫組織染色顯示，苓甘五味薑辛湯可以抑制呼吸道NF-kB的表現。Collagen沉著分析和H.E.染色顯示苓甘五味薑辛湯可以減低呼吸道重塑的情形。在RT-PCR的實驗中，苓甘五味薑辛湯可以抑制IL-10、eotaxin、RANTES mRNA的生成。綜合以上結果顯示苓甘五味薑辛湯作用機轉可能主要是選擇性調控單核球和嗜中性白血球，而不是調控嗜酸性白血球以及和呼吸道重塑（airway remodeling）有關的細胞激素（IL-10、TNF-a、TGF-β）和化學趨化因子（chemokine、eotaxin、RANTES），透過調節Th 1 /Th2免疫反應和抑制轉錄因子NF-kB的活化，來減低呼吸道發炎細胞的浸潤和呼吸道重塑。 Allergic asthma is a chronic airway inflammatory disease, which is characterized by the Th2-bias immirne response, eosinophils infiltration, mast cell activation, epithelial hypertrophy, goblet cell hyperplasia, airway hyperresponsiveness, and excessive mucus secretion in airway. Dermatogoides pteronyssinus (Der p) is one of the most prominent and important species of house dust mite in allergic asthma in Taiwan. In this study, we used repetitive challenge of Der p it instillation in BABL/c mice as a chronic asthmatic animal model, which helped us to evaluate the therapeutic mechanisms of Ling-Gang-Wu-Weng-Jiang-Xin~Tang (LGWWJXT). As a result, we found that LGWWJXT attenuated Der p-induced airway inflammation and remodeling via selectively regulating on macrophages and neutrophils but not eosinophils in the lungs. LGWWJXT also could downregulate TGF-(3 and TNF-a in BALF to inhibit partly eosinophils infiltration and up-regulated IL-12 in BALF to change Th2-bias. In addition, EMSA and immunohistochemistry staining demonstrated LGWWJXT could inhibit NF-kB expression in the lung. In RT-PCR experiment, we also found that LGWWJXT could inhibit IL-10, eotaxin, and RANTES mRNA expression. These results show that the major mechanism of LGWWJXT could be through the regulation of the Thl/Th2 immune response and the inhibition of transcription factor NF-kB. Therefore, the mechanism of LGWWJXT on repetitive Dermatogoides pteronyssinus challenged asthmatic mice model is selectively regulate on macrophages and neutrophils. Its anti-inflammatory activity may modulate Thl/Th2 imbalance as well as reducing NF-kB activation in bronchial epithelium, but also by inhibiting the progressing of airway remodeling.