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標題: | 以α-GalCer誘導小鼠模式探討絨毛膜蛻膜間葉幹細胞在慢性肺疾病中的效果 Study on the Effects of Placenta Choriodecidual-Membrane Derived Mesenchymal Stem Cells on the α-GalCer induced Murine Model of Chronic Lung Disease |
作者: | Cheng-Chiu Tsao 曹正秋 |
指導教授: | 莊雅惠(Ya-Hui Chuang) |
關鍵字: | 慢性肺部疾病,COPD,α-GalCer,間葉幹細胞, Chronic Lung Disease,COPD,α-GalCer,mesenchymal stem cell, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | 慢性肺部阻塞性疾病(chronic obstructive pulmonary disease, COPD)為阻塞型肺部疾病,是一種長期、慢性且無法恢復的呼吸道氣流阻塞,導致氣體無法通暢地進出呼吸道,此疾病包含慢性支氣管炎與肺氣腫兩類病徵。近年來此疾病的盛行率和死亡率在全世界都持續上升,故臨床上對於其機制和治療的研究也更為積極。間葉幹細胞(mesenchymal stem cells)已被證明可以調控多種免疫與發炎相關的疾病,也被認為具有治療肺部發炎疾病之潛力。本實驗室先前的研究發現,給予小鼠α-GalCer活化iNKT細胞後會造成肺部急性發炎反應。在此,我們探討重覆給予小鼠α-GalCer是否造成慢性肺部發炎及其病理變化特徵。此外,我們探討由人類胎盤蛻膜分離培養之間葉幹細胞(placenta choriodecidual- membrane derived mesenchymal stem cells; pcMSCs)對α-GalCer引起之慢性呼吸道發炎是否具有免疫調控之功能。我們以α-GalCer活化BALB/c小鼠之iNKT細胞後發現小鼠肺部的細胞激素IFN-γ及 IL-4升高且浸潤細胞亦增加,確認iNKT細胞在肺部發炎疾病之重要性。接著我們以鼻腔注射重複給予小鼠α-GalCer後顯示這些小鼠有較高的支氣管肺泡沖洗液中浸潤的細胞總數、巨噬細胞、淋巴球(T/NK/NKT細胞)、嗜中性球以及CD8 T細胞。肺臟發炎細胞激素IL-6及TNF-α升高。組織病理分析可發現有細胞浸潤以及肺泡破壞之情形、分泌Mucus的細胞表現量增加,collagen的沉積增加。另外Muc5ac、collagen type 3、MMP-12等基因表現在重複給予α-GalCer的小鼠肺臟均顯著上升。這些慢性發炎反應之特徵與人類COPD相似。此外,體外試驗結果顯示pcMSCs會抑制骨髓培養之巨噬細胞分泌IL-6、TNF-α以及增生的能力。進一步在α-GalCer引起之COPD小鼠模式中給予pcMSCs,結果顯示,與對照組相比,COPD小鼠在接受pcMSCs後支氣管肺泡沖洗液中浸潤的細胞總數及巨噬細胞數均上升,而CD8 T細胞則沒有差異。在組織病理部分,小鼠組織中浸潤的細胞與肺泡破壞程度無顯著差異;然而我們卻發現小鼠在接受pcMSCs後支氣管肺泡沖洗液中CD4 T細胞增加,值得注意的是肺臟發炎細胞激素IL-6及TNF-α顯著降低,組織中黏液細胞減少,此外Muc5Ac表現亦顯著降低。綜合以上實驗,我們以重覆鼻腔注射給予α-GalCer可建立與人類COPD特徵相似之小鼠。另外我們證明pcMSCs可降低COPD的黏液分泌及細胞激素的表現。 Chronic obstructive pulmonary disease (COPD) is characterized by long-term, chronic and irrevrseble airflow limitation. It includes chronic bronchitis and alveolar destruction (emphysema). The mortality and prevalence rate of COPD increases recently and therapeutic approaches are required. Recent studies, which have revealed that mesenchymal stem cells (MSC) contribute to immunomodulation of inflammation disease, are potential therapeutic applications in airway inflammation diseases. Our previous studies demonstrated that activation of iNKT cells by intranasal injection of α-GalCer induced acute airway inflammation. In this study, we investigated whether chronic airway inflammation could be induced in mice by repeated intranasal injection of α-GalCer and we characterized the pathological changes of these mice. We further investigated the effects of placenta choriodecidual-membrane derived mesenchymal stem cells (pcMSCs) on chronic airway inflammation. Our results demonstrated that intranasal administration of α-GalCer induced the activation of iNKT cells by secreting cytokine such as interferon-γ (IFN-γ) and interleukin (IL)–4 and recruited cell infiltration to lung. These results confirmed the importance of activation of iNKT cells in the airway inflammation. By repeated intranasal injection of α-GalCer to mice, we found that increased airway inflammation, including increase in total BALF cell counts, macrophages, lymphocytes (T/NK/NKT cells), neutrophils and CD8 T cells were observed in α-GalCer injected mice. In addition, inflammatory cytokine such as IL-6 and TNF-α in lung increased. In the histopathological study, cell infiltration, alveolar destruction, mucus production and collagen deposition were found in repeatedly intranasal α-GalCer injected mice. Moreover, the level of Muc5ac, collagen type 3 and MMP-12 were also significantly increased. All of these features are similar to human COPD. Next, we showed that pcMSCs could inhibit the IL-6 and TNF-α production and proliferation of macrophage in vitro. We then investigated the effects of pcMSCs using the murine model of COPD. Compared with the control group, BALF total cells and macrophages increased. There was no difference in the number of CD8 T cells while increased CD4 T cells were found. No difference levels of cell infiltration and alveolar destruction were observed in these two groups. Of note, IL-6 and TNF-α production as well as mucus production in tissues and Muc5Ac expression were significantly decreased in COPD mice receiving pcMSCs. In conclusion, we set up a mouse model of COPD by repeated injection of α-GalCer and we demonstrated that pcMSCs may decrease the mucus production and proinflammatory cytokine production of COPD. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/62097 |
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