| Summary: | 碩士 === 國立清華大學 === 生命科學系 === 89 === Cells interact with one another via gap-junctional communication. Gap junctions are clusters of channels located in the cell membrane. Each channel comprises twelve connexin molecules, which belong to a multigene family. Different members of the family contribute to the unique properties of the gap-junctional channels. Because electrical and metabolic signals can exchange between adjacent cells via gap junctions, a variety of biological activities depend on these junctions, including maintenance of cardiovascular functions. The relationship between gap junctions and cardiovascular disease have been investigated and emphasized.
The gap junctions in vascular cells are composed of Cx37, Cx40 and Cx43, and expression of these junctions is affected by a variety of risk factors of atherosclerotic disease. However, the relation between endothelial connexins and atherosclerosis is not clear. This study focused on the effects of atherogenic factors, such as aging and hypertension, on gap junction distribution and connexin expression in rat aortic endothelium. Anti-peptide antisera against Cx40 was generated by affinity purification and characterized by Western blotting and immunocytochemistry. The probe combining with anti-Cx37 and anti-Cx43 antibody was therefore used in immunohistochemistry. Confocal laser scanning microscopy was employed to exam in the effects of aging and hypertension induced by the two kidney one clip model on the connexins in rat aortic endothelium. The results showed that i) gap junction distribution and connexin expression in rat aortic endothelium are age-related and all the three connexins are downregulated at the later stage of life; ii) endothelial Cx43, but not Cx37 and Cx40, was upregulated in hypertension. These findings indicate that gap junction distribution and connexin expression in rat aortic endothelium are altered with aging and hypertension.
英文摘要………………………………………………………………III
名詞縮寫………………………………………………………………V
致謝辭………………..……………………………………………..…VI
目錄………………..……………………………………………..…VII
第一章 緒論…………………………………………………………...1
1.1前言…………..…………………………………………………2
1.2隙連結為細胞之間溝通的管道………………………………….2
1.3隙連結管道的分子組成………………………………………….4
1.4隙連結溝通的調節……………………………………………….6
1.5器官，組織和細胞的隙結……………………………………….9
1.6隙連結和疾病的係…………………………………………...14
1.7隙連結與動脈粥狀硬化………………………………………..16
1.8關於本文……………………………………………………….19
圖1.1………………………………………………………………….20
第二章 材料與方法………………………………………………...21
2.1動物檢體的來源、手術與飼養………………………………22
2.2灌流與固定(Perfusion Fixation)………………………………...22
2. 3免疫組織化學染色(Immunohistochemistry)…………………..23
2. 4共軛焦雷射掃瞄顯微術(Confocal Laser Scanning Microscopy)24
圖2.1………………………………………………………………….27
表2.1……………………………………………………………….28
表2.2……………………………………………………………….28
表2.3……………………………………………………………….28
第三章 抗連接素40抗體的製造，純化和鑑定………………29
3.1前言……………………………………………………………….30
3.2材料與方法….………………………………………………….30
3.3結果……………………………………………………………….37
3.4討論……………………………………………………………….40
圖3.1………………………………………………………………….41
圖3.2………………………………………………………………….42
圖3.3………………………………………………………………….43
圖3.4………………………………………………………………….44
第四章 老化對主動脈內皮細胞的隙連結之影響………….45
4.1前言……………………………………………………………….46
4.2材料與方法……………………………………………………….47
4.3結果……………………………………………………………….47
4.4討論……………………………………………………………….51
圖4.1………………………………………………………………….55
圖4.2………………………………………………………………….56
圖4.3………………………………………………………………….57
表4.1………………………………………………………………….58
第五章 高血壓老鼠主動脈內皮細胞隙連結的表現…………59
5.1前言……………………………………………………………….60
5.2材料與方法……………………………………………………….61
5.3結果……………………………………………………………….62
5.4討論……………………………………………………………….64
圖5.1………………………………………………………………….67
圖5.2………………………………………………………………….68
圖5.3………………………………………………………………….69
圖5.4………………………………………………………………….70
表5.1………………………………………………………………….71
表5.2………………………………………………………………….71
第六章 結論……………………………………………………….…72
參考文獻………….…………………………………………………..75
附錄………………………………………………………………..…..89
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