PHF20介导NF-κB核稳定性和持续活化调控脑出血继发损伤的作用机制研究

Brain edema, inflammation and cell death lead to neurological deterioration after intracerebral hemorrhage (ICH). However, the molecular pathological mechanisms are still unclear. The dearth of clinically effective treatment options makes ICH the least treatable form of stroke. Nuclear factor kappa B (NF-κB) is a transcriptional factor. It is a key regulator of multiple signaling pathways and pathological processes involved in secondary injury of ICH. It has been proved that NF-κB is tightly associated with prognosis of ICH. We firstly indicate that a nuclear protein PHF20 highly expresses in hemorrhage periphery. PHF20 mediates nuclear stability and constitutive activation of NF-κB, which is positively related to activation of microglial cells and expression of cytokines after ICH. Knockdown of PHF20 dramatically increases cell viability after ICH. Therefore, the present study aims to systematically investigate mechanisms of PHF20 on modulating brain edema, inflammation and apoptosis through NF-κB pathway after ICH. Various techniques will be used, including in vivo and in vitro models, gene delivery, immunofluorescence, EMSA, luciferase assay and so on. The results will expand biological functions of PHF20, and will be conducive to improving understand the occurrence and progress of secondary injury of ICH. The study will provide theoretical and experimental basis for designing and verifying new therapeutic strategies against secondary injury of ICH.

脑出血后神经功能恶化与继发水肿、炎性反应和脑细胞凋亡坏死等因素有关，其分子病理机制不清，临床尚缺乏有效治疗方法。核转录因子NF-κB是多种脑出血继发损伤信号转导途径和病理过程的关键调节枢纽，与脑出血预后密切相关。我们率先发现，一种在出血灶周组织高表达的核蛋白PHF20，能够介导NF-κB核稳定性和持续活化，与胶质细胞的活化、炎性因子的表达成正相关，敲除PHF20可显著提高细胞存活率。基于这一发现，本项目拟利用在体、离体模型，结合基因调控、多重免疫荧光、EMSA、报告基因等技术，系统阐述PHF20通过NF-κB信号通路调控脑出血后脑水肿、炎性损伤和神经细胞凋亡的机制。研究结果不仅有望拓展对PHF20蛋白功能的新认识，更有利于完善对脑出血继发损伤发生、发展核心过程的了解，为设计和验证脑出血继发损伤的预防和治疗新策略提供理论基础和实验依据。

脑出血后神经功能恶化与继发水肿、炎性反应和脑细胞凋亡坏死等因素有关，其分子病理机制不清，临床尚缺乏有效治疗方法。核转录因子NF-κB是多种脑出血继发损伤信号转导途径和病理过程的关键调节枢纽，与脑出血预后密切相关。本研究利用了在体、离体模型，结合基因调控、多重免疫荧光、EMSA、报告基因等技术。本研究主要分为三部分1，确立PHF20在ICH损伤不同时相的变化趋势及分布特点；2，阐明PHF20在ICH后继发炎性反应、脑水肿和细胞凋亡中的重要作用；3，阐明PHF20激活NF-κB通路，调控ICH继发损伤的分子作用机制。研究发现脑出血继发损伤发生过程中，PHF20主要定位于神经胶质细胞，脑损伤后表达升高，敲除PHF20后可以显著抑制脑损伤，减轻脑水肿提高神经细胞存活率，降低炎性因子的水平。PHF20主要通过NF-κB/p65的核转运调节炎性反应，本研究基本阐明PHF20蛋白其在脑出血继发损伤中的重要作用和分子机制。我们在研究过程中发现PHF20在多种生物学进程中发挥重要作用，我们以基因芯片为技术载体，通过差异基因分析、信号通路富集和信号网络分析，阐明以PHF20为核心的分子信号网络。本研究系统阐述PHF20通过NF-κB信号通路调控脑出血后脑水肿、炎性损伤和神经细胞凋亡的机制。研究结果不仅有望拓展对PHF20蛋白功能的新认识，更有利于完善对脑出血继发损伤发生、发展核心过程的了解，为设计和验证脑出血继发损伤的预防和治疗新策略提供理论基础和实验依据。