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博碩士論文 etd-0805114-103028 詳細資訊
Title page for etd-0805114-103028
論文名稱
Title
運動蛋白FNDC5/irisin促進內皮細胞血管新生之功能
Exercise protein irisin promotes angiogenesis in endothelial cells
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
63
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2014-07-04
繳交日期
Date of Submission
2014-09-08
關鍵字
Keywords
纖連蛋白III型結構域的蛋白質5、血管新生、棕色脂肪、白色脂肪、內皮細胞
Angiogenesis, white adipose, brown adipose, FNDC5 (Fibronectin type III domain-containing protein 5), endothelial cells
統計
Statistics
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中文摘要
Myokines主要是由肌肉透過運動所分泌出來的cytokines,而在生物體中myokines扮演著重要的角色,例如、血管新生、組織再生和修復、維持健康的身體機能,免疫調節和胚胎發育的影響。而在2012年Boströmet al.發現了新myokine並且稱之為FNDC5。 FNDC5也被發現有誘使白色脂肪細胞棕色化並且進一步消耗熱量來達到減肥、減緩糖尿病病症的能力。在目前的研究中,並沒有明確的指出myokine。 FNDC5是否具有調節血管新生以及傷口癒合的功能,而我們透過大腸桿菌系統生產出來的FNDC5來觀測FNDC5對血管新生的功能並發現在內皮細胞中具有促進細胞增生、遷移與管柱形成之功能。此外,在大鼠主動脈環微血管及Tg(fli-1:EGFP)y1斑馬魚幼苗及傷口癒合中也都有明顯的效果。機制研究表示,在血管內皮細胞中,FNDC5具有促進血管內皮生長因子(VEGF)和血管內皮生長因子受體(VEGFR2)以及下游的p-Erk/Erk與pp38/p38。因此,本研究主要探討FNDC5誘導新血管內皮生長,並期許未來能對糖尿病患者的傷口癒合或是心臟病與中風的治療用藥進行發展。
Abstract
Myokinesis a critical cytokines in organism major secretes from muscle cells after doing exercise. Myokines involved in angiogenesis, tissue regenerate and repair, preserved normal functions of body, affected immune modulation and develop of embryos. Boströmet al. found a new myokine and called FNDC5 in 2012. FNDC5 was founded possessing a ability that can induce White Adipose Tissue becoming Brown Adipose Tissue to lose weight and slow down the symptom of diabetes through increase burning calories. In the resent reports, there is no direct describe that whether FNDC5 can regulates angiogenesis and wound healing. Recombinant FNDC5 (Fibronectin type III domain-containing protein 5) was expressed and purified from E. coli with a molecular weight of 25 kDa. By using cultured endothelial cells, it was found that recombinant FNDC5 stimulated the proliferation, migration and tube formation of endothelial cells. Moreover, FNDC5 promotes vessels sprouting in aortic rings assay and in transgenic Tg(fli-1:EGFP)y1zebrafish. Finally, FNDC5 application enhanced wound healing in diabetic rats. Mechanism studies showed FNDC5 induced the expression of vascular the endothelial growth factor (VEGF)/vascular endothelial growth factor receptor 2 (VEGFR2/FLK-1) pathway and activated the downstream Erk and p38 signaling in endothelial cells. Therefore, this study unveils the novel pro-angiogenic the function and mechanism of FNDC5 in endothelial cells.
目次 Table of Contents
The authorization of oral member for research dissertation ………….i
Acknowledgement.……………………………………………………… .ii
Table of contents………………………………………………………….iii
Abstract in Chinese………...…………………………………………….iv
Abstract in English………………………………………………………..v
Table of contents………………………………………………………….vi
Introduction……………………………………………………………….1
Specific Aims………………………………………………………………6
Materials and Methods…………………………………………………...7
Results…………………………………………………………………….18
Purification of the recombinant FNDC5 protein………………….18
FNDC5 expression in endothelial, liver and muscle cells…………18
The stability of FNDC5 proteins at different temperatures………19
FNDC5 promoted multiple angiogenic processes in endothelial cell…………………………………………………………………….19
FNDC5 effected angiogenesis ex vivo and in vivo…………………20
Identification existed of FNDC5 by membrane, cytoplasm, and nucleus fractions……………………………………………………..21
FNDC5 induces VEGF protein level expression in endothelial cells……………………………………………………………………21
FNDC5 elevated the VEGFR2 expression and VEGFR2 phosphorylation protein level in endothelial cells………………….21
FNSDC5 regulated the VEGF/VEGFR2 through the Erk/p38 signal pathway……………………………………………………………….22
FNDC5 not effect NOS signal pathway in endothelial cells……….23
FNDC5 induced VEGF expression by NFκB in endothelial cells at transcriptional level………………………………………………….23
FNDC5 induces VEGF expression by HIF-1α in endothelial cells at transcriptional level………………………………………………….24
Discussion……………………………………………………………..25
Figures and legends…………………………………………………..29
References……………………………………………………………..52
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