TNF-α 是具有多種功能的細胞激素，在哺乳類動物相關研究中發現可以促使發炎反應、巨噬細胞的吞噬、呼吸爆活性、細胞的分化等等，因此在抵抗病原菌病以及調控免疫機制上佔有重要的角色。目前已經有多種魚類已被發現存在不同數量的TNF-α。在實驗室過去的研究中，在點帶石斑魚上亦選殖到兩個不同的TNF-α，經由LPS的刺激周邊血液細胞後發現兩個TNF-α有著不同的表現量。為了探討此一不同現象的可能成因。本研究將首先以不同的免疫刺激分子刺激魚體探討兩個TNF-α 分子表現的影響並發現兩個TNF-α 對於不同的刺激物具有不同的表現模式。在本研究中，我們發現 (1) 在模擬細菌性 (LPS、 Vibrio amguillarum O1 serotype)或是病毒性的感染(Poly I:C和神經壞死病毒) 時，tnf-α1和tnf-α2的表現量在24小時內都會顯著的上升，並且tnf-α1的表現量高於tnf-α2的表現量。(2) 利用不同的重組蛋白做為免疫刺激物可以發現:經由rgIL-1β、rgIFN-γ刺激時，tnf-α1 的表現量都高於tnf-α2；經由rgCCL4刺激時，tnf-α2 的表現量都高於tnf-α1；經由rgIL-6刺激時，都會使tnf-α1和tnf-α2的表現量下降。(3)以rgTNFα1和rgTNFα2刺激時，可發現其實兩個TNF-α是會互相的影響。本研究的成果顯示tnf-α1和tnf-α2可能存在不同的調控機轉。但詳細內容仍需進一步的研究。

The cytokine, TNF-α, is an important pro-inflammatory cytokine released by several immune cells during infection or tissue damage and is involved in a diverse range of inflammatory, infectious, and malignant conditions. TNF-α also has many functions in mammal, such as induces inflammation, macrophage activation, cell proliferation, apoptosis, and so on. In the previous studies, the functions of teleost TNF-α of fish were simular as mammal, such as increased expression of inflammatory genes. But, more than one TNF-α isoforms were found in teleost fish. Some of this isoforms was different in their amino acid or DNA sequences. In our privious study, we had found 2 orange-spotted grouper TNF genes: gTNF-α1 and gTNF-α2, they were only 38% amino acid identical. And they expressed different mRNA pattern in peripheral blood leukocytes (PBLs) under LPS stimulated, the mechanism is still unknow. In this study, we try to understand the different function in vivo. We analysis the gene expression of immunostimulant stimulated fish by rea-ltime PCR. The results show (1) tnf-α1 has shown a much stronger expression relative to tnf-α2 after LPS, Poly I:C, vibrio and nervous necrosis virus infected. (2) In various recombinated grouper cytokines stimulated fish, tnf-α1 has shown stronger expression than tnf-α2 after injection rgIL-1β and rgIFN-γ, but tnf-α2 seems to be a more significant factor than tnf-α1 after rgCCL4 injection. However, tnf-α1 and tnf-α2 both down expression after rg-IL-6. (3)TNF-α1 and TNF-α2 have shown the cross activation in each other. Overall, the results indicate that maybe tnf-α1 has more important role on innate immunity, and tnf-α2 is a helper to induce tnf-α1expression.

方苡帆，應用重組介白素-6於點帶石斑魚之抗病機轉研究，國立成功大學生物科技研究所碩士論文，2013。

吳思穎，點帶石斑魚 (Epinephelus coioides) 二腫瘤壞死因子-α (TNF-α1、TNF-α2) 其基因、蛋白結構及表現差異性之探討，國立成功大學生物科技研究所碩士論文，2011。

林伯峰，點帶石斑魚兩腫瘤壞死因子-兩腫瘤壞死因子壞死、TNF-壞死因子蛋白功能性分析之探討，國立成功大學生物科技研究所碩士論文，2012。

蔡侑翰，Cathepsin L在點帶石斑魚上的功能性分析，國立成功大學生物科技研究所碩士論文，2012。

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