RIP3在necroptosis信号通路中的胞内定位

The intracellular localization of RIP3 in necroptosis signaling pathways

目的 研究necroptosis信号通路中受体相互作用蛋白3（RIP3）的胞内定位.方法 SD大鼠原代皮质神经元细胞培养12天,zVAD 20 μmol/L预处理30 min,TNFα 30 ng/ml处理不同时间,Western blot检测胞质、胞核中RIP3蛋白水平,结合免疫荧光观察RIP3的胞内定位.结果 随着TNFα作用时间的延长,胞质、胞核中RIP3蛋白水平均呈上升趋势,胞质中RIP3蛋白水平在8 h达高峰,随后下降,胞核中RIP3蛋白水平在12 h达高峰,即在12 h出现核转位的高峰（P＜0.05）.结论 正常细胞中RIP3存在于胞质,necroptosis时发生核转位.

Objective To investigate the intracellular localization of receptor - interacting protein 3 （ RIP3 ） in necroptosis and its function in this signal passage. Methods Primary cerebrocortical neurons were cultured for 12 days, followed by pretreatment with zVAD for 30 mn to block apoptosis and treatment with TNFα for different durations. Then, the cytoplasmic and nuclear protein levels of RIP3 were analyzed by Western blot and the intracellular localization of RIP3 was observed by immunofluorescenee. Results In signaling pathways of necroptosis, the cytoplasmic protein level of RIP3 gradually decreased with prolonged TNFα exposure, with a gradual corresponding rise in the nuclear protein level and its peak of nuclear translocation at 12 h of TNFα treatment （ P 〈 0.05 ）. Conclusion In normal ceils, RIP3 is norreally located in the cytoplasm, while nuclear translocation takes place in necroptosis.