- 學位論文
探討球薑酮對於免疫細胞的調控與對氣喘小鼠呼吸道發炎現象之影響
The modulatory effects of zerumbone on the immune effector cells and airway inflammation in a murine model of asthma
摘要
球薑酮 (zerumbone)為紅球薑 (Zingiber zerumbet Smith)的主要成分，文獻記載其具有抗發炎、抑制腫瘤生成、抗氧化以及促進細胞凋亡的效果。而本實驗目的在於進一步探討球薑酮是否對於免疫調節細胞如：樹突細胞、T細胞具有調節活化及分化的功能，進而達到治療及預防呼吸道過敏反應的效果。在細胞實驗部分，利用球薑酮給予樹突細胞觀察細胞激素分泌情形及樹突細胞表面分子的表現，以及在T細胞給予球薑酮觀察細胞增生、細胞激素分泌情形，並且藉由混合淋巴細胞反應 (mix lymphocyte reaction, MLR)觀察球薑酮受到刺激的樹突細胞是否具有調節T細胞分化的功能。在動物實驗方面，分別以卵白蛋白 (ovalbumin, OVA)致敏小鼠氣喘動物模式給予球薑酮餵食，觀察是否具有治療及預防的效果。實驗中測量其呼吸道阻力、血清抗體、肺沖洗液細胞激素及細胞種類，脾臟細胞之細胞激素分泌情形。實驗結果顯示球薑酮可以直接降低樹突細胞分泌介白質素10 (interleukin-10, IL-10)及介白質素12 (interleukin-12, IL-12)及表面分子CD40的表現，並且可以直接促進T細胞增生；在MLR反應發現可以促進T細胞增生並且使丙型干擾素 (interferon-gamma, IFN-γ)分泌量有增加的趨勢。在動物實驗結果顯示不論在治療或是預防的模式下，餵食球薑酮皆可抑制因呼吸道過度反應造成的呼吸道阻力增加的現象，並且血清IgE分泌受抑制、IgG2a產量提高，肺沖洗液內發炎性細胞激素IL-5、IL-13、IL-10、發炎介質eotaxin、KC的分泌量減少、IFN-γ分泌量增加，肺沖洗液內嗜酸性白血球及嗜中性球數量減少、脾臟細胞分泌TNF-α、IL-4、IL-5、IL-13與IL-10的能力皆降低。綜合以上實驗結果我們推論：球薑酮可能藉由促進Th1細胞生成增加IFN-γ分泌量，以達到抑制Th2細胞分化，從而使因氣喘反應造成的呼吸道發炎現象獲得改善。因此認為將來可能可以利用球薑酮來改善及預防氣喘疾病。
並列摘要
Zerumbone is the main in gredient of Zingiber zerumbet Smith. There is a wide applications of zerumbone. It is applied to reduce a variety of inflammatory diseases and inhibits tumor cell formation. In addition, studies have found that zerumbone has the antioxidation effect and can promote cell apoptosis. In our study, we investigated whether zerumbone possessed immunoregulatory effect on immunol cells, such as dendritic cells (DCs) and T cells. By the way, we evaluated the preventitive and therapetic effects of zerumbone in asthmatic animal model. First, DCs were treated with lipopolysaccharide (LPS) and zerumbone. The levels of cytokine secretion and surface molecules expression were observed. Furthermore, we detected the proliferation and cytokines secrection on T cells which were treated with anti-CD3/anti-CD28 and zerumbone. Then mixed lymphocyte reaction (MLR) was used to investigate whether zerumbone-treated DCs could regulate the differentiation of T cells. In animal experiments, an OVA-induced asthmatic animal model was used to observe whether zerumbone treatment could alleviate or prevent the development of asthma by measuring airway resistance, serum antibody, airway inflammation and cytokines production from BALF and splenocytes. Our data demonstrated that zerumbone inhibited the production of IL-10 and IL-12 from DCs challenged with LPS plus zerumbone. Zerumbone also suppressed the expression of CD40 on DCs. Besides, the proliferation of T cells was markedly enhanced by treating with anti-CD3/anti-CD28 and zerumbone. In addition, zerumbone-treated DCs significantly enhanced T cells proliferation and increased interferon-gamma (IFN-γ) secretion. Furthermore, we evaluated the preventive and therapeutic effects of zerumbone in asthmatic model. The results showed that zerumbone decreased IgE product, airway hyperresponsiveness (AHR), airway inflammation and Th2 cytokines secrection. Such regulation from zerumbone might be through an IFN-γ-dependent pathway. In conclusion, this study makes us further understand the regulatory roles of zerumbone, and ultimately results in zerumbone-based drug to prevent and treat atopic asthma.