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標題: | 忍冬葉桑寄生萃取物在過敏疾病治療及免疫調節機制之探討 Immune modulatory effect of a pure compound Isolated from Taxillus lonicerifolius var. lonicerifolius in murine model of asthma |
作者: | Hsin-Yi Huang 黃馨儀 |
指導教授: | 江伯倫 |
關鍵字: | 忍冬葉桑寄生萃取物,氣喘,樹突細胞,Th1細胞免疫反應, Taxillus lonicerifolius var. lonicerifolius,asthma,Dendritic cell,Th1 immunity,IL12, |
出版年 : | 2007 |
學位: | 碩士 |
摘要: | 氣喘是一種慢性呼吸道發炎性疾病,在全世界無論是在已發展或發展中的國家,都是非常普遍的疾病。造成氣喘發炎反應的原因,主要是因為T輔助型細胞的發育偏向Th2細胞型免疫反應;其產生的細胞激素以IL-4、IL-5和IL-13為主。氣喘發炎反應就是由在這些細胞激素與體內過敏原專一性IgE抗體的主導下,啟開一連串的連鎖反應所造成的。其主要臨床症狀包括了氣管收縮、氣管過度反應和氣管發炎反應。已經有強烈的證據證明了,直接給予Th1細胞激素或是利用刺激的方式增加Th1細胞激素的產生,例如給予IFN-α,IFN-γ,IL-12 或CpG-motif oligodexynucleotide皆可降低Th2呼吸道的發炎反應,進而達到治療氣喘動物模式的目的。而樹突細胞是一個專業的抗原呈獻細胞,具有可以調控T細胞分化走向的能力。忍冬葉桑寄生(T. lonicerifolius var. lonicerifolius)屬於桑寄生科(Loranthaceae),鈍果桑寄生屬(Taxillus),是台灣特有種的桑寄生植物。在本實驗中,我們取用由長庚大學天然藥物所呂彥禮助理教授所提供的忍冬葉桑寄生萃取物,研究其對於樹突細胞的影響。實驗發現,給予忍冬葉桑寄生萃取物可增加樹突細胞IL-12p40,IL-12p70和IL-10的表現。同時也可增加樹突細胞CD80、CD86、MHCI和MHCII等細胞表面分子的表現,並抑制樹突細胞的吞噬能力,刺激樹突細胞的成熟。進一步,利用anti-TLR-4抗體做中和反應的實驗,則發現IL-12p40的產生會受到抑制。因此認為,忍冬葉桑寄生萃取物可透過TLR-4的訊息傳遞以刺激樹突細胞的反應。另外,利用受刺激修飾後的樹突細胞與T細胞共同培養,發現可增加T細胞增生能力以及分泌IFN-γ的能力。總結於in vitro的結果,忍冬葉桑寄生萃取物具有調節免疫反應的能力。進一步利用I.P.注射忍冬葉桑寄生萃取物與OVA抗原混和液,建立小鼠動物模式,以探討忍冬葉桑寄生萃取物對OVA專一性細胞免疫反應走向的影響。結果發現,給予忍冬葉桑寄生萃取物作為佐劑的組別,小鼠血清中的OVA抗原專一性IgG2a有顯著性的增加;而OVA抗原專一性IgE有顯著性的抑制。雖然在脾臟細胞培養實驗中,IFN-γ並無顯著性的增加,但是IL-4和IL-5細胞激素皆有顯著性的降低。這顯示以忍冬葉桑寄生萃取物作為佐劑,確實具有刺激BALB/c小鼠抗原專一性免疫反應傾向Th1細胞免疫反應發展。由於這些發現,我們認為忍冬葉桑寄生萃取物是一個具有可以治療氣喘動物模式的化合物。在未來,我們將進一步利用忍冬葉桑寄生萃取物對氣喘小鼠做治療的研究。 Allergic asthma is one of the most important Th2 lymphocyte-mediated chronic inflammatory airway disease worldwide, both in the developing and developed countries. In the patients of allergic asthma, it is well characterized by skewed polarization of helper T cell development toward type 2 T helper cell with increased production of interleukin 4 (IL-4) and IL-5. Asthma is characterized by airway eosinophils inflammation, mucus hypersecretion, bronchial hyperreactivity, the presence of high levels of allergen-specific IgE. There is strong evidence for the therapeutic effect of type 1 T helper-related cytokines or reagents, such as IFN-α, IFN-γ, IL-12 or CpG-motif oligodexynucleotide on airway inflammation. Dendritic cell are the most potent antigen presenting cells for priming the naïve T helper cells. Taxillus lonicerifolius var. lonicerifolius is one species of Loranthaceae in Taiwan. One pure compound (1400041), kindly providing by Pro. Leu,. isolated from the fresh T. lonicerifolius var. lonicerifolius was used in this study. In this study, we found stimulating BM-DC with 30μg/ml 1400041 could enhance production of IL-12 p40, IL-12 p70 and IL-10, as well as enhance cell surface expression of CD80 (B7.1), CD86 (B7.2), MHCI and MHCII molecule. In addition, the capacity for endocytosis was suppressed in BM-DC treated with 1400041. And, neutralization with antibody against Toll-like receptor(TLR)-4 inhibited the 1400041-induced production of IL-12 p40, suggest a crucial role for TLR-4 in signaling DC upon incubation with 1400041. In addition, treatment of BM-DC with 1400041 resulted in enhanced T cell-stimulatory capacity and increased T cell secretion of IFN-γ. Taken together, our in vitro data demonstrated that this 1400041 can effectively promote the activation and maturation of DC, suggesting that 1400041 may possess a potential in regulating immune responses. To test the effect of 1400041 in vivo, we further examined the effects of this compound on antigen-specific antibody and cytokine production. Immunization with ovalbumin (OVA) / 1400041 showed that the anti-OVA IgG2a levels were significantly increased and the anti-OVA IgE levels were significantly decreased compared with that of OVA-alone group. While 1400041 had no significantly effect on the anti-OVA IgG1 levels and induced the secretion of IFN-g, it inhibited the secretion of IL-4 and IL-5. The findings demonstrate that 1400041 could be used as adjuvant to induce Th1 immunity in BALB/c mice. In the future, we aim to study the therapeutic effect and immune modulatory mechanism of 1400041 in murine model of asthma. We expect this compound might be used as novel therapeutic reagent for asthma treatment. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/30824 |
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顯示於系所單位: | 口腔生物科學研究所 |
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