石斑鱼RIP1在细胞凋亡和NF-κB激活中的作用机制

当机体或组织被病原感染或受到炎症因子刺激的时候，细胞会通过各种信号途径激活NF-κB、IRFs或者丝裂原活化蛋白激酶（MAPKs）等来上调表达一些抗凋亡蛋白或炎症因子来保护细胞和机体；相反地，细胞也可以通过凋亡或者坏死来清除受损的细胞或“同归于尽”的清除侵入细胞的病原体。大量的研究表明受体相关蛋白激酶（RIP）家族作为一类基本的细胞内“传感器”决定了多种细胞压力的结果。迄今为止，鱼类中RIP功能报道较少。我们前期工作中成功克隆了石斑鱼RIP1基因。本项目拟在此基础上，利用基因克隆、荧光定量PCR、免疫荧光、免疫共沉淀、免疫印迹、报告基因、点突变、基因过表达等技术，开展如下研究：1）RIP1表达模式和细胞定位；2）RIP1在NF-κB激活中的作用；3）RIP1在细胞凋亡中的作用 。研究结果将为理解鱼类RIP1相关信号通路的基本功能和RIP1相关疾病的潜在免疫调控机制提供新的认识。

When the body or tissue is pathogenic or stimulated by inflammatory factors, cells will up-regulate the expression of some anti-apoptotic proteins or inflammatory factors to protect the cells and the body by a variety of signaling pathway activation of NF-κB, IRFs or mitogen-activated protein kinase (MAPKs) and so on; On the contrary, the cells can also remove the damaged cells or "die" to remove the invasive pathogens by the apoptosis or necrosis. Many studies have shown that receptor-associated protein kinase (RIP) families as a class of basic intracellular "sensors" determine the results of multiple cell pressures. But so far, little related information of RIP1 has been reported in fish. In our previous research, a RIP1 sequence from grouper (EcRIP1) was identified. Overexpression of EcRIP1 inhibited NF-κB, ISRE and IFN-βactivation. In this project, the mechanisms of EcRIP1in apoptosis and activation of NF-κB will be thoroughly studied by using molecular and cellular techniques, including gene cloning, real-time quantitative PCR, immunofluorescence, Co-Immunoprecipitation, western blot, luciferase reporter assay, single base substitution and gene overexpression/silencing. This investigation will focus on: 1)The expression patterns and intracellular localization of EcRIP1. 2)The regulations of NF-κB activation by EcRIP1. 3) The mechanism of EcRIP1in apoptosis. The findings will provide a new understanding of the basic functions of the RIP1-related signaling pathway and the underlying immune regulatory mechanisms of RIP1-related diseases.