MLKL信号途径在调控鱼类鳃细胞坏死致烂鳃中的作用和机制研究

鱼类鳃非常重要，但容易遭受损伤（烂鳃），导致摄氧量和采食量下降、生长受阻、死亡率高，严重威胁产业安全。鳃损伤常与细胞坏死有关，但鱼类细胞坏死的调控机制不清楚。MLKL信号途径在调控人和鼠细胞坏死中起关键作用，但调控机制在不同物种间存在差异，在鱼上缺乏研究。因此，本项目拟在鉴定草鱼鳃MLKL、RIPK3、RIPK1、TRIF等信号分子的基础上，综合利用siRNA、基因重组、融合荧光蛋白、流式细胞、荧光标记等技术揭示RIPK1、TRIF结合RIPK3形成坏死体调控MLKL形成寡聚体转位细胞膜破坏脂膜结构或阳离子通透性介导鳃细胞坏死的作用和机制；进一步通过体内和体外研究揭示（TRIF、RIPK1）/RIPK3/MLKL信号途径在肌醇抑制鱼类鳃坏死中的作用和机制。通过系列研究揭示（TRIF、RIPK1）/RIPK3/MLKL信号途径在鱼类鳃细胞坏死中的作用和机制，有重要的学术意义和较大的应用价值。

Fish gills are very important, but they are vulnerable to injury (gill rot), resulting in decreases of oxygen intake and feed intake, depression of growth and caused high mortality, which seriously threaten the safety of aquaculture industry. Gill injure is often associated with cell necrosis, but the regulatory mechanisms is unclear in fish. MLKL signaling pathway plays a key role in regulating cell necrosis in humans and mice, but the regulatory mechanism is different among different species, and there is little research on fish. Therefore, on the basis of identifying signal molecules such as MLKL, RIPK3, RIPK1 and TRIF in the gills of grass carp, this project aims to reveal the roles and mechanisms of RIPK1 or TRIF binding RIPK3 to form necrosis bodies and thus regulates the formation of MLKL oligomer translocation to the plasma membrane, destroying membrane structure or cationic permeability mediating of gill cell necrosis by using siRNA, gene recombination, fusion fluorescent protein, flow cytometry and fluorescence labeling techniques. Furthermore, we will do in vivo and in vitro studies to reveal the roles and mechanisms of (TRIF or RIPK1)/RIPK3/MLKL signaling pathway in the inhibition of gill necrosis by myo-inositol. Together, we will reveal the roles and mechanisms of (TRIF or RIPK1)/RIPK3/MLKL signaling pathway in the gill cell necrosis of fish, which has important academic significance and great application value.