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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 李啟明(Chii-Ming Lee),江伯倫(Bor-Luen Chiang) | |
dc.contributor.author | Wan-Tseng Hsu | en |
dc.contributor.author | 徐莞曾 | zh_TW |
dc.date.accessioned | 2021-06-16T06:51:25Z | - |
dc.date.available | 2014-10-15 | |
dc.date.copyright | 2014-10-15 | |
dc.date.issued | 2014 | |
dc.date.submitted | 2014-07-23 | |
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/57558 | - |
dc.description.abstract | 研究背景及目的
心臟衰竭為一牽涉多重致病機轉之心臟末期病變,其成因以缺血性心肌病變最為常見。儘管許多醫療策略致力於治療心肌梗塞續發之心臟衰竭,其死亡率與長期預後仍待改善。文獻指出心肌梗塞後的免疫發炎反應會導致心臟收縮功能惡化及重塑作用,適時調控心肌發炎反應可防止心臟衰竭。另一方面,心臟衰竭末期病患主要治療方式之一為心臟移植,但移植接受者的長期存活率卻受限於心臟植體動脈硬化;此病變為一種加速型、瀰漫性的粥狀硬化,冠狀動脈因內膜增生而阻塞,導致心肌缺氧。心臟植體動脈硬化的致病機轉主要為異體免疫反應,但目前臨床上使用的抗排斥藥物並無法有效防治。因此,不論是針對心肌梗塞續發之心臟衰竭或心臟移植植體動脈硬化,發展新的免疫調控策略實屬迫切。 骨髓間葉幹細胞 (bone marrow-derived mesenchymal stem cells, BM-MSCs) 經試管內及體內實驗證明具有調控免疫系統的潛能。本論文假設骨髓間葉幹細胞可有效治療上述免疫發炎反應失調的心血管疾病。本論文研究架構是利用骨髓間葉幹細胞的兩種治療策略,一是自體幹細胞移植法,二是內源性幹細胞驅動法,將研究區分為兩部份,分別於植體動脈硬化與心肌梗塞的豬隻模式,測試其免疫調控效能,並闡述潛在的機轉。 第一部份:移植自體骨髓間葉幹細胞治療植體動脈硬化 骨髓間葉幹細胞經文獻證實可誘發調節性T細胞 (regulatory T cell, Treg) 以調控異體免疫反應。目前研究最詳盡的Treg包括CD4+CD25+Foxp3+ Treg 與第一型調節性T細胞 (T regulatory type 1 cell, TR1);前者主要靠Foxp3決定其免疫調控能力,後者則仰賴IL-10為免疫調控的介質。近期研究發現TR1中某一亞群因具有同時分泌IL-10與IFN-γ 的特性,而被命名為「類第一型調節性T細胞」(TR1-like cell)。本實驗室利用豬隻股動脈同種異體移植模式,證實局部注射自體骨髓間葉幹細胞可有效抑制植體動脈硬化,並增加植體血管組織中的IL-10+ 和IFN-γ+ 細胞。本論文更進一步闡明,在豬隻周邊血液淋巴細胞混合培養反應 (mixed lymphocyte reaction, MLR) 中,骨髓間葉幹細胞可誘發具有IL-10+IFN-γ+CD4+ 表現型的T細胞,此群T細胞於試管內可抑制異體抗原所引起之淋巴球增生反應,因而具有TR1-like cell的特質。然而,TR1-like cell不同於骨髓間葉幹細胞,在於無法分泌前列腺素E2 (prostaglandin E2 , PGE2),且其免疫抑制效力僅為骨髓間葉幹細胞的25%,若補足予PGE2則可進一步加強TR1-like cell的免疫抑制作用。本論文更證實骨髓間葉幹細胞在MLR系統中誘發的TR1-like cell,於活體內無法單獨減緩植體血管內膜的增生,需於移植術後長期補充PGE2,方能再現骨髓間葉幹細胞治療植體動脈硬化的療效。因此,PGE2與TR1-like cell,為骨髓間葉幹細胞調控植體動脈硬化的必要媒介,本研究的成果可作為後續發展心臟移植病患免疫抑制療程的基礎。 第二部份:驅動內源性骨髓間葉幹細胞治療心肌梗塞 除了侵入性的抽取骨髓幹細胞再注射回體內治療標的,破壞骨髓內stromal cell-derived factor 1 (SDF-1) 對CXCR4+ 幹細胞的滯留作用,亦為驅動幹細胞進入周邊血液循環的核心機制。本研究使用之幹細胞驅動劑TG-0054 (學名burixafor, 布利沙福),為CXCR4的小分子競爭性拮抗劑,已於人體第二期臨床試驗證實可有效使CD34+ 幹細胞自骨髓移入周邊血液,本研究進一步探勘 TG-0054 於豬隻體內對骨髓間葉幹細胞的驅動效能。首先,本研究利用細胞貼附性篩選法和CD271表面抗原分選法,分別培養出plastic adherent-MSC (PA-MSC) 與CD271+ cell derived-MSC (CD271-MSC),作為TG-0054驅動幹細胞的指標。經過細胞貼附性、分化能力與細胞表面抗原的鑑定,證實自TG-0054注射後的豬隻血液分離出的PA-MSC與CD271-MSC符合間葉幹細胞的定義,並利用混合淋巴球培養系統,闡明PA-MSC與CD271-MSC可抑制淋巴球增生與降低發炎性細胞激素產生。再者,本研究利用經心導管冠狀動脈氣球阻塞術建立豬隻心肌梗塞模式,於心肌梗塞後第3天與第7天注射TG-0054 (n = 12) 或生理食鹽水 (n = 10)。結果顯示,TG-0054治療組的左心室射出分率 (left ventricular ejection fraction, LVEF),於心肌梗塞後12週 (46 ± 12%) 仍維持與基準值 (47 ± 10%) 相當;經過12週測量的左心室射出分率變化值 (ΔLVEF),治療組與控制組之間達到顯著差異 (PΔLVEF = 0.014),代表TG-0054的治療,可以防護左心室收縮功能免受心肌梗塞損傷而惡化。除此之外,於心肌梗塞後7天,發炎相關基因包括TNF-α、IL-1β與IL-6,在TG-0054治療組的心肌表現量顯著低於生理食鹽水控制組。若分離培養出TG-0054驅動至血液的自體CD271-MSC,按照TG-0054的療程靜脈注射至實驗豬隻,於心肌梗塞後7天,TNF-α、IL-1β與IL-6的心肌表現量亦顯著降低。同時,於心肌梗塞後7天、6週與12週,TG-0054治療組的血液發炎因子包括TNF-α、IL-1β與IL-6亦顯著低於生理食鹽水控制組,此效果亦可藉由注射TG-0054驅動至血液的自體CD271-MSC而達成。因此,本研究推論TG-0054治療可藉由減緩心肌梗塞後局部與全身性的發炎免疫反應以維持左心室功能,此免疫調控作用可能來自於TG-0054驅動的骨髓間葉幹細胞。此研究成果可作為後續發展心肌梗塞治療策略的基礎。 結論 本論文證實,不論以自體幹細胞移植或是驅動內源性幹細胞的方式,骨髓間葉幹細胞於體內皆具有免疫調控作用,可藉由分泌 PGE2 與誘發TR1-like cell 以減緩植體動脈硬化,亦可降低心肌梗塞引發之心肌與全身性的發炎反應而防止左心室收縮功能惡化。應用本研究的成果,可為植體動脈硬化或心肌梗塞引發之缺血性心臟病,發展以免疫調控為基礎的治療策略。 | zh_TW |
dc.description.abstract | Background
Despite recent advances in reperfusion strategies and pharmacotherapy, myocardial infarction (MI) remains a leading cause of morbidity and mortality. In patients that survive MI, the subsequent left ventricular (LV) remodeling and heart failure (HF) remain major health problems. Inflammation and immune responses are vital in adverse remodeling and HF. On the other hand, heart transplantation is one of the established treatment modalities for end-stage HF. However, transplant arteriosclerosis (TA) remains the major limitation of long-term graft survival in heart transplantation despite the advances in immunosuppressants. Therefore, investigations to modulate the immuno-inflammatory responses are mandatory to improve prognosis of both ischemic HF patients and transplant recipients. Bone marrow-derived mesenchymal stem cells (BM-MSCs) are known for their immunomodulatory functions. The aim of this doctoral dissertation is to investigate the immunomodulatory potential of BM-MSCs in the treatment of TA and post-MI HF. The dissertation consists of two parts: the first part is to apply autologous BM-MSCs in alleviating TA and the second part is to mobilize endogenous BM-MSCs for prevention of post-MI HF. Part I: Application of BMSCs in Transplant Arteriosclerosis In this part of study, we investigated the therapeutic potential of autologous BM-MSCs on TA in a porcine model of femoral arterial transplantation. MSCs or saline were injected into the soft tissue surrounding the arterial grafts immediately post anastomosis. Four weeks after transplantation, neointimal formation increased significantly in untreated allografts compared with the MSC-treated grafts as assessed by intravascular ultrasound (maximum luminal area stenosis: 40 ± 12% versus 18 ± 6%, p < 0.001). Grafts harvested at 4 weeks showed dense perivascular lymphocyte infiltration accompanied by significant intimal hyperplasia in the untreated but not in the MSC-treated allografts. Serial angiographic examination showed that all of the untreated allografts became occluded at the 8th week whereas the majority of the MSC-treated grafts remained patent at the 12th week post transplantation (n = 12 each group, p < 0.001). Quantitative PCR analysis revealed that Foxp3 expression was comparable between the untreated and the MSC-treated groups. However, expression of interleukin-10 (IL-10), interferon- | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T06:51:25Z (GMT). No. of bitstreams: 1 ntu-103-D98421001-1.pdf: 5202344 bytes, checksum: f8146f6cdf4a60d8291235a0022551aa (MD5) Previous issue date: 2014 | en |
dc.description.tableofcontents | 目錄
口試委員會審定書 i 誌謝 ii 目錄 iii 圖目錄 v 表目錄 vii 中文摘要 viii 英文摘要 xi 博士論文正文 第一章 緒論 1 壹、間葉幹細胞簡介 1 貳、免疫發炎反應於缺血性心臟疾病扮演的角色 13 參、幹細胞應用於缺血性心臟疾病之治療策略 18 肆、研究目的 25 第二章 研究方法及材料 27 壹、第一部份:移植自體骨髓間葉幹細胞治療植體動脈硬化 一、骨髓間葉幹細胞之特性與功能 27 二、移植自體骨髓間葉幹細胞治療植體動脈硬化的效果與機制 32 三、統計分析 35 貳、第二部份:驅動內源性骨髓間葉幹細胞治療心肌梗塞 一、幹細胞驅動劑移動骨髓間葉幹細胞至周邊循環的效能 35 二、驅動骨髓間葉幹細胞治療心肌梗塞的效果與潛在機轉 37 三、統計分析 41 第三章 結果 42 壹、第一部份:移植自體骨髓間葉幹細胞治療植體動脈硬化 一、骨髓間葉幹細胞之特性與功能 42 二、移植自體骨髓間葉幹細胞治療植體動脈硬化的效果 43 三、移植自體骨髓間葉幹細胞治療植體動脈硬化的潛在機轉 44 四、結果簡述 49 貳、第二部份:驅動內源性骨髓間葉幹細胞治療心肌梗塞 一、幹細胞驅動劑TG-0054的間葉幹細胞驅動效能 50 二、幹細胞驅動劑TG-0054治療心肌梗塞的效果 52 三、幹細胞驅動劑TG-0054治療心肌梗塞的潛在機轉 52 四、結果簡述 54 第四章 討論 56 第五章 展望 72 第六章 論文英文簡述 77 第七章 參考文獻 92 第八章 圖表 140 第九章 附錄發表論文 164 | |
dc.language.iso | zh-TW | |
dc.title | 骨髓間葉幹細胞治療心血管疾病之免疫調控潛能
第一部份:移植自體骨髓間葉幹細胞治療植體動脈硬化 第二部份:驅動內源性骨髓間葉幹細胞治療心肌梗塞 | zh_TW |
dc.title | Immunomodulatory Potential of Bone Marrow-Derived Mesenchymal Stem Cells in Cardiovascular Diseases
Part I: Application of Autologous MSCs in Transplant Arteriosclerosis Part II: Mobilization of Endogenous MSCs in Myocardial Infarction | en |
dc.type | Thesis | |
dc.date.schoolyear | 102-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 司徒惠康(Huey-Kang Sytwu),洪士杰(Shih-Chieh Hung),林泰元(Thai-Yen Ling) | |
dc.subject.keyword | 骨髓間葉幹細胞,植體動脈硬化,心肌梗塞,免疫調控,第一型調節性 T 細胞,CXCR4 拮抗劑, | zh_TW |
dc.subject.keyword | mesenchymal stem cell,transplant arteriosclerosis,myocardial infarction (MI),immunomodulation,T regulatory type 1 cells,CXCR4 antagonist, | en |
dc.relation.page | 164 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2014-07-24 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 臨床醫學研究所 | zh_TW |
顯示於系所單位: | 臨床醫學研究所 |
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