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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 謝學真(Hsyue-Jen Hsieh) | |
dc.contributor.author | Chang-Ting Chen | en |
dc.contributor.author | 陳長廷 | zh_TW |
dc.date.accessioned | 2021-05-20T20:55:58Z | - |
dc.date.available | 2021-09-02 | |
dc.date.available | 2021-05-20T20:55:58Z | - |
dc.date.copyright | 2011-08-03 | |
dc.date.issued | 2011 | |
dc.date.submitted | 2011-07-29 | |
dc.identifier.citation | Abe K, Kimura H (1996) The possible role of hydrogen sulfide as an endogenous neuromodulator. J Neurosci 16: 1066-1071.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/10016 | - |
dc.description.abstract | 硫化氫是近年來受到許多研究關注的細胞內氣體傳導分子(gasotransmitter),在心血管系統中,硫化氫產生許多的保護作用。層流剪力(laminar shear stress)亦是提供心血管保護作用的因子之一,且剪力與硫化氫所誘發的訊息傳導多有類似之處。然而,目前關於剪力對硫化氫生成量的影響,乃至於硫化氫在剪力誘導的訊息傳導方面的知識非常有限,因此,本論文將對此課題作深入討論,主要討論方向有兩點:第一,剪力是否增加細胞內硫化氫生成量;第二,硫化氫對剪力誘導的訊息傳導之影響。
由於在心血管系統中血液流動所產生的剪力最主要作用在血管內皮細胞(vascular endothelial cells)上,因此,本論文主要以人類臍帶靜脈內皮細胞(HUVECs)以及牛大動脈內皮細胞(BAECs)為研究對象。實驗發現,層流剪力在作用30分鐘之後,會使BAECs內的硫化氫生成速率提升5倍。然而,三種可能參與硫化氫生成的酵素:cystathionine-γ-lyase (CSE)、cystathionine-β-synthase (CBS)與3-mercapto-sulfurtransferase (3-MST)的蛋白質量在剪力作用30分鐘後皆無明顯變化,顯示剪力可能透過增加上述硫化氫生成酵素之活性來提高硫化氫之生成量。至於探討硫化氫對剪力誘導的訊息傳導之影響方面,主要使用CSE、CBS與3-MST的siRNA分別抑制其表現量,再觀察剪力刺激下細胞內訊息傳導的變化。結果發現pAkt在受剪力刺激後所被誘導的程度會被上述三種siRNA所抑制,可以推論剪力作用所造成的pAkt上升,部分是由於細胞內硫化氫增高所導致。 綜合以上研究結果,可以發現剪力所產生的心血管保護作用可能部份來自於內皮細胞中硫化氫濃度升高的效應,且CSE、CBS以及3-MST可能是負責硫化氫生成主要的酵素。此外,實驗結果更進一步指出pAkt可能在硫化氫相關的心血管保護作用中扮演重要的角色。 | zh_TW |
dc.description.abstract | Hydrogen sulfide (H2S) is a novel gasotransmitter in cardiovascular system, it renders multiple cardioprotective effects. Laminar shear stress is also a cardioprotective factor in cardiovascular system, which shares many similarities in downstream signal transduction pathways with H2S. However, knowledge about the regulation of H2S production by shear stress and the role of H2S in shear-induced signal transduction is still limited. In the present study, efforts were made to investigate the effect of shear stress on H2S production. Furthermore, the role of H2S in shear-induced cell signaling is also discussed.
In cardiovascular system, shear stress exerted by blood flow mainly affects vascular endothelial cells (ECs). Thus, human umbilical vein endothelial cells (HUVECs) and bovine aortic endothelial cells (BAECs) are used in present study. After laminar shear stress stimulation for 30 minutes, H2S production rate is enhanced by 5 fold in BAECs. Three enzymes possibly involved in H2S production—cystathionine-γ-lyase (CSE), cystathionine-β-synthase (CBS), and 3-mercapto-sulfurtransferase (3-MST)—are not significantly affected by shear stress in terms of protein and mRNA level, implying the possibility that shear stress enhaces H2S production rate by increasing CSE, CBS, and 3-MST enzyme activities. In order to study the role of H2S in shear-induced cell signaling, CSE, CBS, and 3-MST were knocked down by siRNAs before HUVECs were stimulated by shear stress. Results show that shear-induced Akt phosphorylation is inhibited by CSE, CBS, and 3-MST siRNAs. In conclusion, shear-induced cardioprotective effect might be partly a result of increased H2S concentration in ECs. Futrther, shear stress induces Akt phosphorylation in a H2S-dependent way. | en |
dc.description.provenance | Made available in DSpace on 2021-05-20T20:55:58Z (GMT). No. of bitstreams: 1 ntu-100-R98524022-1.pdf: 3764128 bytes, checksum: 5f0701746d2b891cf03258b6014c30c1 (MD5) Previous issue date: 2011 | en |
dc.description.tableofcontents | 致謝 I
中文摘要 III Abstract V 目錄 VII 圖目錄 X 表目錄 XIII 縮寫與符號說明 XIV 中英文名詞對照 XVII 第1章 文獻回顧 1 1.1. 常見心血管疾病 1 1.1.1. 動脈粥狀硬化 (Atherosclerosis) 1 1.1.2. 高血壓 (Hypertension) 6 1.2. 血管內皮細胞與剪力 8 1.2.1. 血管內皮細胞 8 1.2.2. 剪力對血管內皮細胞之影響 12 1.3. 硫化氫與其生理功能 17 1.3.1. 硫化氫在生理上的特性 17 1.3.2. 硫化氫之生成 18 1.3.3. 硫化氫對心血管系統之影響 24 1.4. Akt訊息傳導 30 1.4.1. Akt於心血管系統扮演之角色 30 1.4.2. 剪力對Akt訊息傳導之調控 34 1.5. 一氧化氮 (Nitric Oxide)訊息傳導 35 1.5.1. 一氧化氮於心血管系統之生成與扮演之角色 35 1.5.2. 剪力對一氧化氮訊息傳導之調控 39 1.6. ERK訊息傳導 40 1.6.1. ERK於心血管系統扮演之角色 40 1.6.2. 剪力對ERK訊息傳導之調控 44 1.7. 研究動機與目的 45 第2章 實驗材料、儀器、原理與方法 47 2.1. 實驗材料 47 2.1.1. 細胞培養及流動實驗 47 2.1.2. 西方墨點轉印法 48 2.1.3. siRNA轉染 49 2.1.4. 硫化氫測定 49 2.1.5. 反轉錄 49 2.1.6. 即時定量聚合酶連鎖反應 (Real-time quantitative PCR) 50 2.1.7. 免疫沉澱(Immunoprecipitation) 50 2.1.8. 實驗耗材 50 2.2. 實驗儀器 52 2.3. 實驗原理與方法 53 2.3.1. 初級人類臍帶靜脈內皮細胞培養 53 2.3.2. 人類臍帶靜脈內皮細胞繼代培養 54 2.3.3. 牛主動脈內皮細胞繼代培養 54 2.3.4. 內皮細胞繼代培養於玻片 54 2.3.5. 流動實驗 55 2.3.5.1. 流動室之設計 55 2.3.5.2. 流動實驗之設計與流程 60 2.3.6. 細胞內蛋白質抽取 63 2.3.7. 蛋白質含量測定 63 2.3.8. 西方墨點法 63 2.3.9. 細胞內total RNA之抽取 64 2.3.10. 反轉錄 65 2.3.11. 即時定量聚合酶連鎖反應 (Real-time quantitative PCR) 65 2.3.11.1. DyNAmo™ Flash SYBR® Green qPCR 65 2.3.11.2. TaqMan® Gene Expression Assay 66 2.3.12. siRNA轉染 67 2.3.13. 免疫沉澱 67 2.3.14. 硫化氫測定 67 2.3.15. 量化與統計方法 70 第3章 實驗結果與討論 71 3.1. 剪力對硫化氫生成速率之調控 71 3.2. 剪力對硫化氫合成酵素之影響 73 3.2.1. 剪力對硫化氫合成酵素mRNA之調控 73 3.2.2. 剪力對硫化氫合成酵素蛋白質表現之調控 76 3.3. Calmodulin對硫化氫生成酵素之調控 78 3.4. 硫化氫對內皮細胞之影響 81 3.5. 剪力誘導之訊息傳導與硫化氫之關係 86 3.5.1. 硫化氫清除劑對剪力誘導之訊息傳導之影響 86 3.5.2. 硫化氫生成酶siRNA對剪力誘導之訊息傳導之影響 89 3.6. 綜合討論 103 第4章 結論 109 4.1. 結論 109 4.2. 未來研究方向 111 參考文獻 113 | |
dc.language.iso | zh-TW | |
dc.title | 剪力對內皮細胞中硫化氫生成之調控與訊息傳導之影響 | zh_TW |
dc.title | Regulation of H2S Production and H2S-Dependent Signal Transduction in Endothelial Cells by Shear Stress | en |
dc.type | Thesis | |
dc.date.schoolyear | 99-2 | |
dc.description.degree | 碩士 | |
dc.contributor.coadvisor | 王寧(Danny Ling Wang) | |
dc.contributor.oralexamcommittee | 裘正健(Jeng-Jiann Chiu),翁炳孫(Being-Sun Wung) | |
dc.subject.keyword | 剪力,硫化氫,內皮細胞,心血管保護,Akt, | zh_TW |
dc.subject.keyword | shear stress,hydrogen sulfide,endothelial cells,cardioprotective,Akt, | en |
dc.relation.page | 129 | |
dc.rights.note | 同意授權(全球公開) | |
dc.date.accepted | 2011-07-29 | |
dc.contributor.author-college | 工學院 | zh_TW |
dc.contributor.author-dept | 化學工程學研究所 | zh_TW |
顯示於系所單位: | 化學工程學系 |
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