Brg1正调控STAT1/PUMA通路介导的细胞凋亡在肝缺血再灌注损伤中的作用研究

项目名称： Brg1正调控STAT1/PUMA通路介导的细胞凋亡在肝缺血再灌注损伤中的作用研究

项目编号： No.81501695

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 葛缅

作者单位： 中山大学

项目金额： 18万元

中文摘要： 肝缺血再灌注损伤（HIRI）是肝脏缺血手术病人不良预后的主要原因，细胞凋亡在HIRI发展中扮演重要角色，机制复杂仍未完全阐明。PUMA是促凋亡关键因子，STAT1是新近发现与凋亡有关核转录因子，但STAT1与PUMA关系仍不明确；Brg1是染色质重塑复合物重要亚基，因增强转录效能。我们前期研究证明小鼠HIR引起明显细胞凋亡；预实验发现HIR后肝Brg1、STAT1和PUMA表达上调而P53变化不显著，在Brg1基因敲除小鼠HIRI模型中这些现象明显改善；据此，我们设想Brg1正调控STAT1/PUMA介导的细胞凋亡是肝缺血再灌注损伤的重要机制。本项目拟用Brg1基因敲除鼠HIRI和肝细胞缺氧复氧模型，结合质粒转染过表达、基因沉默和Chip等技术，通过在体实验和细胞学研究论证三者的相互关系及对细胞凋亡的影响，以验证这一假说，阐明HIRI的机制，为防治HIRI提供新的干预靶点及理论依据。

中文关键词： 肝缺血再灌注；细胞凋亡；表观遗传；基因转录；Brg1

英文摘要： Hepatic Ischemia-reperfusion Injury (HIRI) is one of the main contributors to poor prognosis of patients who underwent hepatic surgery. The mechanisms of HIRI remains to be elucidated and effective therapeutic intervention are still being called for. Apoptosis plays an important role in HIRI. PUMA is a pro-apoptotic factor. STAT1 is a transcription factor which act as a pro-apoptotic signal. Brg1 associated chromatin remodeling complex can regulate the transcription and expression of multiple genes, and participate in many pathophysiological process. Our previous research has shown in the liver tissue of rats underwent HIRI, Brg1, STAT1 and PUMA were upregulated, along with elevated mRNA levels, poor liver function and pathological changes. In Brg1 KO mice, however, the liver function and pathological changes were significantly restored, and the protein expression and mRNA levels of Brg1, STAT1 and PUMA were all lowered. Therefore, we hypothesize that the regulation of STAT1/PUMA pathway-induced apoptosis by Brg1 is the key to HIRI. Our current research intend to apply certain genetic modification techniques such as adeno-virus transfection, Chip analysis and dual luciferase reporter assay, to establish in vitro and in vivo ischemia-reperfusion/hypoxia-redox injury model, and elucidate the mechanisms of Brg1 in regulating STAT1/PUMA induced apoptosis and the impact of this pathway on HIRI to provide new therapeutic target to HIRI.

英文关键词： hepatic ischemia reperfusion;apoptosis ;epigenetic;genetic transcription;Brg1