臺灣博碩士論文加值系統

創傷弧菌(Vibrio vulnificus)，是一種嗜鹽的革蘭氏陰性菌，外型微彎曲且有鞭毛能移動，適合生存在10 – 31℃的環境下，通常被發現在溫暖的淺水海域中，由於此菌引起的感染速度很快，感染後死亡率高達50%，所以近年來已經有受到越來越多臨床醫師開始關注。有幾種族群是比較容易受到創傷弧菌感染後，會有嚴重情況出現，包含肝病及慢性肝炎的患者、酗酒、有慢性疾病等免疫功能不好的族群。台灣四面環海B型肝炎患者較多且一般民眾喜歡吃海鮮類的食物，因此創傷弧菌的防治值得我們注意。
先前實驗室的研究指出，創傷弧菌可以利用他們所產生的一種細胞外毒素(H毒素)造成宿主細胞的死亡。本研究要分析創傷弧菌與巨噬細胞進行反應後，H毒素是否會影響創傷弧菌的移動性及巨噬細胞的吞噬作用及菌體於巨噬細胞的存活率，藉此明白H毒素對創傷弧菌感染的重要性。
研究結果顯示H毒素會讓創傷弧菌主動向巨噬細胞移動，而且當巨噬細胞吞入創傷弧菌後，創傷弧菌可以利用H毒素幫助對抗巨噬細胞的吞噬，增加菌體在巨噬細胞內的存活率。

Vibrio vulnificus is a halophilic, motile, curved Gram-negative bacterium living in ocean. Infection caused by this bacterium progresses very fast, and high mortality rate up to 50%. In recent years, more and more clinicians have begun to pay attention to study this bacterium. Several groups experience serious complications after being infected with Vibrio vulnificus, including alcoholics, liver disease and chronic hepatitis, or immune insufficiency. Taiwan is surrounded by the sea, and many people prefer to eat seafood food, so prevention and treatment of Vibrio vulnificus infection deserves our attention.
Previous studies have shown that Vibrio vulnificus can use one of their extracellular toxins (H toxin) to kill host cell. This study was to determine the effect of H toxin of Vibrio vulnificus on bacterial mobility and phagocytosis by macrophages. The results showed that H toxin plays an important role in increasing the mobility of Vibrio vulnificus and the bacterial survival rate after phagocytosis

目錄
中文摘要 I
Abstract II
目錄 III
圖目錄 V
縮寫表 VII
第一章 緒論 1
1-1 創傷弧菌(Vibrio vulnificus) 1
1-1-1 感染的症狀 3
1-1-1-1 原發性敗血症(primary septicemia) 3
1-1-1-2 次發性敗血症(secondary septicemia) 4
1-1-1-3 急性腸胃炎(gastrointestinal diseases) 4
1-1-2 致病因子(virulence factors) 4
1-1-2-1 酯多糖體(lipopolysaccharide, LPS) 5
1-1-2-2 莢膜多醣體(capsular polysaccharide, CPS) 5
1-1-2-3 RTX 毒素(RTX toxin) 5
1-1-2-4 鞭毛(flagella) 6
1-1-2-5 線毛(pili) 6
1-1-2-6 細胞溶解素(cytolysin) 7
1-1-2-7 金屬蛋白酶(metalloprotease) 7
1-1-2-8 鐵採集系統(iron acquisition systems) 8
1-1-2-9 溶血素(hemolysin) 8
1-2 巨噬細胞(macrophage) 8
1-2-1 吞噬作用 9
1-3 研究動機 9
第二章 材料與方法 12
2-1 材料與藥品 12
2-1-1 創傷弧菌(Vibrio vulnificus) 12
2-1-2 細胞 12
2-1-3 培養基 12
2-1-3-1 細菌培養基 12
2-1-3-2 細胞培養基 13
2-1-4 抗生素 13
2-1-5 溶液 13
2-2 實驗步驟 14
2-2-1 細菌培養 14
2-2-1-1 固態培養 14
2-2-1-2 液態培養 14
2-2-2 細胞培養 14
2-2-2-1 細胞繼代 14
2-2-2-2 細胞計數的方法 15
2-2-2-3 細胞培養於6孔盤 15
2-2-2-4 細胞培養於96孔盤 16
2-2-3 菌數調整 16
2-2-4 吞噬實驗 16
2-2-5 創傷弧菌移動的分析 17
2-2-6 細胞轉染 17
第三章 結果 18
3-1 吞噬實驗 18
3-1-1 創傷弧菌受巨噬細胞吞噬的分析 18
3-1-2 創傷弧菌附著於巨噬細胞的分析 18
3-1-3 創傷弧菌存活率分析 19
3-2 胞外鈣離子對巨噬細胞殺菌的影響 19
3-3 創傷弧菌移動的分析 20
3-4 創傷弧菌H毒素基因轉染進入HeLa細胞的情形 20
第四章 討論 22
第五章 結論 26
參考文獻 27
圖 34

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