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研究生:羅子鈞
研究生(外文):Tzu-Chun Lo
論文名稱:巨噬細胞對斑馬魚的心臟再生作用
論文名稱(外文):Roles of Macrophages on Zebrafish Heart Regeneration
指導教授:謝錦源
指導教授(外文):Dennis Jine-Yuan Hsieh
學位類別:碩士
校院名稱:中山醫學大學
系所名稱:醫學檢驗暨生物技術學系碩士班
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2017
畢業學年度:105
語文別:中文
論文頁數:69
中文關鍵詞:巨噬細胞心肌細胞再生作用
外文關鍵詞:MacrophagescardiomyocyteRegeneration
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心肌梗塞(MI)通常導致心肌細胞死亡,在人類發生心肌梗塞導致心肌細胞死亡,體內的免疫系統修復受損的心肌細胞會產生出疤痕。並且人類損傷後的心肌細胞不能顯著再生。在新生小鼠中,心肌細胞可以在出生後7天內(P7)後MI完全再生而無瘢痕。然而,具有巨噬細胞缺陷的新生小鼠不能再生心肌並形成纖維化瘢痕,導致心臟功能和血管生成減少。這是發現巨噬細胞提供必要的信號來觸發新生小鼠心臟的血管生成和再生。相比之下,斑馬魚顯示出心臟終其一生皆可再生。在本研究中,使用Cl2MDP-L和Metronidazole介導的巨噬細胞耗竭模型,實驗中澄清巨噬細胞和斑馬魚心臟再生的關係。為了分析再生過程,在心肌損傷後不同天數收集斑馬魚,固定和切片。通過使用H&E染色和Masson trichrome stain 摻入來檢測心肌細胞纖維化程度。證明了巨噬細胞的有無能夠觸發斑馬魚心臟的血管生成和再生過程中扮演著重要的角色。
Myocardial infarction (MI) often leads to cardiomyocyte death, and human beings are unable to regenerate significantly after an acute injury. In neonatal mice, cardiomyocytes can regenerate completely without scarring following MI before postnatal day 7 (P7). However, neonatal mice with macrophages deficiency are unable to regenerate myocardia and form fibrotic scars, resulting in reduced cardiac function and angiogenesis. It was findings that macrophages provide necessary signals to trigger angiogenesis and regeneration of the neonatal mouse heart. In contrast, zebrafish shows a remarkable ability for cardiac regeneration. In this study, using a macrophages-depletion model by CL2MDP-L and mediated by MTZ-induced cardiomyocytes apoptosis, we clarify the roles of macrophages on zebrafish heart regeneration. In order to analyze the regenerative process, the hearts were collected, fixed at different days after MTZ-induced cardiomyocytes apoptosis. By using H&E, Masson’s trichrome serial heart sections to visualized fibrosis. We suggested that macrophages are able to trigger regeneration of the zebrafish cardiomyotes.
中文摘要..............................................v
英文摘要.............................................vi
第一章、緒論..........................................1
第一節、斑馬魚模式動物................................1
第二節、斑馬魚胚胎發育................................2
第三節、造血作用......................................5
第四節、斑馬魚造血系統................................6
第五節、斑馬魚心臟發育................................7
第六節、巨噬細胞......................................8
第七節、心肌細胞凋亡..................................9
第八節、微脂粒 (Liposome).............................10
第九節、Dichloromethylenediphosphonic acid disodium salt..................................................11
第十節、Metronidazole.................................13
第十一節、組織專一性表達NTR的基因轉殖魚做法...........14
第十二節、實驗動機....................................15
第二章、實驗材料和方法................................19
第一節、斑馬魚的飼育與胚胎收集........................19
第二節、Liposome之製備................................21
第三節、Neutral red ..................................22
第四節、組織切片染色..................................23
第五節''統計...........................................27
第三章、實驗結果......................................28
第四章、討論..........................................36
第五章、參考文獻......................................38
第六章、圖表說明......................................44
第七章、附錄..........................................66
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