自从受体相互作用蛋白3（RIP3）介导细胞坏死的调控机制被发现之后，对于细胞坏死信号通路的研究就迅速成为生物医学领域的前沿热点。已有实验表明RIP3并不能作为理想的药物靶点，因此对于细胞坏死过程中调控蛋白的发现及其机制的深入研究并协助治疗药物的开发具有重要意义。系统生物学和质谱技术的发展为人们实现系统和定量研究生物学过程提供了重要保障。本项目拟针对TNF-α诱导小鼠L929细胞的细胞坏死信号通路，利用免疫沉淀的方法纯化相关的蛋白复合物，结合Group-DIA质谱技术对相应的复合物蛋白进行鉴定及定量分析；基于得到的质谱数据，利用系统生物学理论对相应的细胞坏死信号网络模型进行构建，并结合实验验证定量探讨细胞坏死的动力学机制及预测发现细胞坏死过程中的未知调控蛋白，该研究将为协助临床上与细胞坏死相关疾病的治疗制定最优策略及发现更多潜在的药物靶点提供一定的理论依据。

Since the regulatory mechanism of receptor-interacting protein 3 (RIP3) -mediated cell necrosis has been found, the study of necrosis signaling pathway has rapidly become a hotspot in the biomedical field. Experiments has shown that RIP3 can not be used as an ideal drug target, so it is important for the discovery of therapeutic drugs to study the proteins and their function in the necrosis process. The development of systems biology and mass spectrometry technology provides guarantees for systematic and quantitative studying the biological processes. In this study, based on the TNF-α-induced necrosis signaling pathway of L929 mouse cells , we first employ the immunoprecipitation method to purify the related protein complexes and analysis the complexes by using Group-DIA mass spectrometry. Then we apply the systems biology theory to construct the corresponding RIP3-mediated necrosis signaling network model according to the former analysis results. Combining with experiments, the kinetic mechanism of necrosis will be eventually investigated and the unknown regulatory proteins will be predicted. This study will provide some theoretical basis for developing clinical optimal treatment strategies and the potential drug targets discovery of the diseases associated with necrosis.