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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 陳美如 | |
dc.contributor.author | Hsueh-Hsia Wu | en |
dc.contributor.author | 吳雪霞 | zh_TW |
dc.date.accessioned | 2021-06-08T07:00:51Z | - |
dc.date.copyright | 2009-09-15 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-05-26 | |
dc.identifier.citation | Bonacorsi, S. and Bingen, E., 2005. Molecular epidemiology of Escherichia coli causing neonatal meningitis. Int J Med Microbiol 295, 373-81.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/26125 | - |
dc.description.abstract | 大腸桿菌為引起新生兒腦膜炎主要的革蘭氏陰性細菌,外膜蛋白OmpA是大腸桿菌外膜的主要蛋白質成分之ㄧ,在大腸桿菌與宿主細胞間相互作用上扮演重要的角色。為研究外膜蛋白OmpA在大腸桿菌侵入星狀細胞時的重要性,我們發現OmpA+大腸桿菌菌株E44,E105以及E109相對於OmpA-菌株E91和E111具有高約10至15倍的附著和侵入C6神經膠瘤細胞的能力。細胞內肌動蛋白(actin)的重组,酪氨酸蛋白激酶(protein tyrosine kinase)和磷脂醯肌醇-3激酶(phosphoinositide 3-kinase)的活化都參與在此侵入的過程中。在試管內E44感染C6細胞株的實驗或C57BL/6小鼠經由顱內注射E44的感染,都會刺激星狀細胞過度表現其活化指標膠質纖維酸性蛋白(glial fibrillary acidic protein),同時也會刺激細胞表現發炎性介質,如環氧酵素2(cyclooxygenase 2)以及誘導型一氧化氮合成酶(nitric oxide synthase 2)等。C57BL/6小鼠經由顱內注射E44感染後36小時內會死亡,相對地,如果細菌注射時同時給予純化的重組OmpA蛋白,則能使小鼠有80%的存活率。染色觀察小鼠腦部組織切片,發現重組OmpA蛋白可以抑制細菌感染後所引起的星狀細胞被激活以及嗜中性白血球的浸潤。總言之,本研究的結果顯示大腸桿菌藉由外膜蛋白OmpA附著進而侵入星狀細胞,在細菌感染初期所引起的腦部傷害扮演著重要的角色,同時也暗示其在大腸桿菌腦膜炎治療與預防上可能的應用。 | zh_TW |
dc.description.abstract | Escherichia coli is the major Gram-negative bacteria pathogen in neonatal meningitis. Outer membrane protein A (OmpA) is a conserved major protein in the E. coli outer membrane and is involved in several host-cell interactions. To characterize the role of OmpA in the invasion of astrocytes by E. coli, we investigated OmpA-positive and OmpA-negative E. coli strains. Outer membrane protein A + E44, E105, and E109 strains adhered to and invaded C6 glioma cells 10- to 15-fold more efficiently than OmpA-negative strains. Actin rearrangement, protein tyrosine kinase, and phosphoinositide 3-kinase activation were required for OmpA-mediated invasion by E. coli. In vitro infection of C6 cells and intracerebral injection into mice of the E44 strain induced expression of the astrocyte differentiation marker glial fibrillary acidic protein and the inflammatory mediators cyclooxygenase 2 and nitric oxide synthase 2. After intracerebral infection with E44, all C57BL/6 mice died within 36 hours, whereas 80% of mice injected with E44 premixed with recombinant OmpA protein survived. Astrocyte activation and neutrophil infiltration were reduced in brain tissue sections in the mice given OmpA. Taken together, these data suggest that OmpA-mediated invasion plays an important role in the early stage of E. coli-induced brain damage, and that it may have therapeutic use in E. coli meningitis. | en |
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dc.description.tableofcontents | 中文摘要 I
英文摘要 II 專有名詞縮寫表 III 目錄 V 第一章 前言 1 1. 中樞神經 1 1.1 細胞組成 1 1.1.1 小膠質細胞(Microglia cells) 1 1.1.2 星狀細胞(Astrocytes) 2 1.2 中樞神經的感染 3 1.2.1 細胞素與趨化素(Cytokines and Chemokines) 3 1.2.2環氧酵素2(Cyclooxygenase 2) 4 1.2.3 誘導型一氧化氮合成酶(Nitric oxide synthase 2) 4 2. 細菌性腦膜炎 5 3.大腸桿菌 5 3.1 毒力因子(Virulence factors) 6 3.1.1 黏附素(Adhesins) 6 3.1.2 蛋白質水解酶(Proteases) 6 3.1.3鐵離子擷取系統(Iron acquisition systems) 7 3.1.4分泌系統(Secretory systems) 7 3.1.5 其他 8 3.2 外膜蛋白OmpA(Outer membrane protein A) 8 4. 研究目的 9 第二章 實驗材料及方法 10 1. 細胞培養 10 2. 化學藥品、細菌菌株及培養方法 10 3. 細菌附著與侵入實驗 10 4. 細胞內細菌的存活與細菌感染後C6細胞的生長曲線 11 5. 電子顯微鏡檢查 11 6. SDS-PAGE與西方墨點法 12 7. 螢光免疫染色與共軛焦顯微鏡檢查 12 8. His-OmpA與His-enolase蛋白的表現與純化 12 9. 動物實驗 13 10. 統計 14 第三章 結果 15 1. 大腸桿菌K1菌株藉由外膜蛋白OmpA附著與侵入C6細胞 15 2. 大腸桿菌藉由外膜蛋白OmpA侵入C6細胞的過程會引起肌動蛋白重組,酪氨酸蛋白激酶與磷脂醯肌醇-3激酶的活化 16 3. 大腸桿菌藉由外膜蛋白OmpA侵入C6細胞後引起細胞的活化 17 4. 小鼠大腦內注射OmpA+大腸桿菌會活化星狀細胞,並且使星狀細胞表現COX-2和NOS-2 18 5. 大腦內注射OmpA+大腸桿菌會造成C57BL/6小鼠的死亡 19 6. 重組OmpA蛋白抑制大腸桿菌侵入C6細胞以及細胞的活化 19 7. 重組OmpA蛋白可保護C57BL/6小鼠避免因大腸桿菌感染引起的死亡 20 8. 重組OmpA蛋白可抑制在動物大腦內因大腸桿菌感染所引起的星狀細胞活化以及COX-2和NOS-2的表現 20 9. 結論 21 第四章 討論 22 1. 外膜蛋白OmpA是影響大腸桿菌對C6細胞的附著與侵入能力的主要因素 22 2. 大腸桿菌侵入C6細胞時所需要的訊息傳遞路徑以及侵入細胞後的結果 23 3. 外膜蛋白OmpA是大腸桿菌附著,侵入進而活化不同細胞的重要蛋白 24 4. 重組OmpA蛋白將有助於發展新的治療方法,以改進細菌性腦膜炎的預後 26 5. 未來研究方向 26 第五章 圖表 28 表一、不同大腸桿菌附著與侵入C6細胞的能力 28 表二、大腸桿菌菌株感染後小鼠腦部存留的細菌量 29 圖一、大腸桿菌菌株附著與侵入C6細胞的能力 30 圖二、以穿透式電子顯微鏡觀察大腸桿菌菌株侵入C6細胞 32 圖三、大腸桿菌E44菌株侵入C6細胞時所涉及的訊息傳遞路徑 33 圖四、大腸桿菌藉由外膜蛋白OmpA侵入C6細胞引起GFAP過度表現 34 圖五、OmpA+大腸桿菌感染小鼠腦部促使星狀細胞活化並表現COX-2和NOS-2 35 圖六、不同大腸桿菌菌株腦部感染後小鼠的存活觀察 37 圖七、His-OmpA抑制大腸桿菌侵入C6細胞及GFAP的過度表現 38 圖八、顱內感染大腸桿菌時,His-OmpA具有保護小鼠存活的能力 39 圖九、顱內感染大腸桿菌時,His-OmpA能抑制星狀細胞的活化以及COX-2和NOS-2的表現 40 圖十、顱內感染大腸桿菌時,His-OmpA的存在可以減少腦部嗜中性白血球的浸潤 41 圖十一、大腸桿菌藉由外膜蛋白OmpA附著進而侵入星狀細胞後所引發相關分子表現的可能機轉 42 第六章 參考文獻 43 | |
dc.language.iso | zh-TW | |
dc.title | 外膜蛋白OmpA在大腸桿菌侵入及活化星狀細胞扮演角色之探討 | zh_TW |
dc.title | Study on the OmpA-Mediated Escherichia coli Invasion and Activation of Astrocyte | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 陳振陽,賈景山,李建國,賴信志,楊沂淵 | |
dc.subject.keyword | 大腸桿菌,外膜蛋白A,星狀細胞,膠質纖維酸性蛋白,環氧酵素2,誘導型一氧化氮合成酶, | zh_TW |
dc.subject.keyword | Astrocyte,Cyclooxygenase 2,Escherichia coli,Glial fibrillary acidic protein,Nitric oxide synthase 2,Outer membrane protein A, | en |
dc.relation.page | 48 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2009-05-27 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 微生物學研究所 | zh_TW |
顯示於系所單位: | 微生物學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
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ntu-98-1.pdf 目前未授權公開取用 | 2.05 MB | Adobe PDF |
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