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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/17402

Title: 點帶石斑重組interleukin-1β蛋白對海水魚免疫反應之影響
The effects of grouper recombinant interleukin-1β in the immune response of marine fish
Authors: Siang-Hua Chen
陳湘華
Contributors: NTOU:Institute of Marine Biology
國立臺灣海洋大學:海洋生物研究所
Keywords: 細胞介白素-1;免疫;重組蛋白
interleukin-1;immune;recombinant protein
Date: 2005
Issue Date: 2011-07-04
Abstract: 摘要 本研究之目的在利用基因重組方式以E. coli生產出recombinant interleukin-1β (rIL-1β),再經過活體以及離體實驗,檢驗其生理活性,並探討其對石斑魚以及黑鯛免疫反應之影響。實驗主要分為(1) 石斑魚rIL-1β之合成與純化(2) 細菌感染對於石斑魚IL-1β基因表現量之影響 (3)rIL-1β對於石斑魚以及黑鯛頭腎白血球超氧陰離子強度之影響(4)rIL-1β對遭dexamethasone抑制頭腎白血球超氧陰離子強度之石斑魚以及黑鯛之影響(5)活體注射rIL-1β對遭受Vibrio anguillarum病原菌感染之石斑魚之影響。 經低劑量E. coli浸潤後,石斑魚的頭腎及脾臟之IL-1β表現量有顯著增加,但高劑量E. coli組,表現量卻低於低劑量E. coli組。在胸腺部分,低劑量E. coli組的IL-1β表現與控制組無顯著差異,高劑量E. coli組IL-1β表現量低於低劑量E. coli組,控制組胸腺IL-1β顯著高於頭腎與脾臟之表現。 在離體實驗中,當rIL-1β濃度達1×10-9 M時可顯著提升黑鯛及石斑魚之頭腎白血球超氧陰離子強度。而在dexamethasone與rIL-1β相加實驗中,當rIL-1β添加濃度達1×10-7 M時才可抑制dexamethasone對石斑魚以及黑鯛之頭腎白血球超氧陰離子強度之影響。 在活體注射實驗中,低劑量與高劑量rIL-1β加上病原菌感染之實驗組,可能因為過量之rIL-1β導致嚴重發炎反應,使得其死亡率高於僅以病原菌感染之實驗組。離體實驗中,所有病原菌感染組其頭腎白血球超氧陰離子強度皆高於控制組。額外添加rIL-1β之離體實驗,在控制組以及高劑量rIL-1β注射加上病原菌感染組,有顯著增加白血球超氧陰離子強度之作用。反之,單純感染組以及低劑量rIL-1β加上病原菌感染組則無增強作用。 本研究經離體實驗證明點帶石斑的rIL-1β確具活性,但其在活體的應用上,尚需進一步實驗檢驗其在產業應用之可利用性。
Abstract The objectives were to produce grouper (Epinephelus coioides) recombinant interleukin-1β (rIL-1β) and to investigate the effects of rIL-1β in the immune response of marine fish. Treatment with low dose of E. coli resulted in an increase in the expression of IL-1β in the head kidney and spleen of grouper. However, the expressions in high dose group were lower as compared to the low dose group. The expression in control and low dose group had no significant difference, whereas the expression in the high dose group was lower than that of the low dose group. In vitro experimental trials showed that a concentration of rIL-1β at 1×10-9 M could promote WBC superoxide anion production efficiently. Furthermore, when the concentration of rIL-1β reached to 1×10-7 M, it could recover the inhibitory effect of dexamethasone on WBC superoxide anion strength. In vivo experiments revealed high mortality rates in the Vibrio anguillarum infected group even in the presence of low or high dose rIL-1β. This may be due to tissue impairment induced by over-dose of rIL-1β. The superoxide anion strength was higher in the control as compared to any dose of the infected groups. After vivrio infection, different concentrations of rIL-1β were separately added to the extracted WBC from different groups. A concentration of 1×10-5 M could promote the superoxide anion strength alone in the control and high dose rIL-1β plus infection group but not in the infection only or low dose rIL-1β plus infection group. Thus the present study demonstrated the bioactivity of rIL-1β by in vitro experiments. However, further studies are needed to show the protective effects of rIL-1β in the infectious marine fish.
URI: http://ethesys.lib.ntou.edu.tw/cdrfb3/record/#G0M93340010
http://ntour.ntou.edu.tw/ir/handle/987654321/17402
Appears in Collections:[海洋生物研究所] 博碩士論文

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