Ge Mian, Yao Weifeng, Yuan Dongdong, Zhou Shaoli, Chen Xi, Zhang Yihan, Li Haobo, Xia Zhengyuan, Hei Ziqing
Department of Anesthesiology, Third Affiliated Hospital, Sun Yat-sen University, Guangzhou, Guangdong 510630, China.
Department of Anesthesiology, Li Ka Shing Faculty of Medicine, The University of Hong Kong, Hong Kong, China.
Cell Death Dis. 2017 Jun 1;8(6):e2841. doi: 10.1038/cddis.2017.236.
Cytoprotective gene heme oxygenase 1 (HO-1) could be induced by nuclear factor E2-related factor 2 (Nrf2) nuclear translocation. The purpose of this study was to determine the role of Brahma-related gene 1 (Brg1), a catalytic subunit of SWI2/SNF2-like chromatin remodeling complexes, in Nrf2/HO-1 pathway activation during hepatic ischemia-reperfusion (HIR). Our results showed that hepatic Brg1 was inhibited during early HIR while Brg1 overexpression reduced oxidative injury in CMV-Brg1 mice subjected to HIR. Moreover, promoter-driven luciferase assay showed that overexpression of Brg1 by adenovirus transfection in AML12 cells selectively enhanced HO-1 gene expression after hypoxia/reoxygenation (H/R) treatment but did not affect the other Nrf2 target gene NQO1. Furthermore, inhibition of HO-1 by the selective HO-1 inhibitor zinc protoporphyria could partly reverse the hepatic protective effects of Brg1 overexpression while HO-1-Adv attenuated AML12 cells H/R damage. Further, chromatin immunoprecipitation analysis revealed that Brg1 overexpression, which could significantly increase the recruitment of Brg1 protein to HO-1 but not NQO1 promoter, was recruited by Nrf2 to the HO-1 regulatory regions in AML12 hepatocytes subjected to H/R. In conclusion, our results demonstrated that restoration of Brg1 during reperfusion could enhance Nrf2-mediated inducible expression of HO-1 during HIR to effectively increase antioxidant ability to combat against hepatocytes damage.
细胞保护基因血红素加氧酶1(HO-1)可由核因子E2相关因子2(Nrf2)的核转位诱导。本研究的目的是确定SWI2/SNF2样染色质重塑复合物的催化亚基婆罗门相关基因1(Brg1)在肝缺血再灌注(HIR)期间Nrf2/HO-1通路激活中的作用。我们的结果表明,在早期HIR期间肝脏中的Brg1受到抑制,而Brg1过表达可减轻接受HIR的CMV-Brg1小鼠的氧化损伤。此外,启动子驱动的荧光素酶测定表明,在AML12细胞中通过腺病毒转染过表达Brg1可在缺氧/复氧(H/R)处理后选择性增强HO-1基因表达,但不影响其他Nrf2靶基因NQO1。此外,选择性HO-1抑制剂锌原卟啉对HO-1的抑制可部分逆转Brg1过表达的肝脏保护作用,而HO-1腺病毒可减轻AML12细胞的H/R损伤。此外,染色质免疫沉淀分析显示,Brg1过表达可显著增加Brg1蛋白与HO-1启动子的结合,但不与NQO1启动子结合,在接受H/R的AML12肝细胞中,Nrf2可将Brg1募集至HO-1调控区域。总之,我们的结果表明,再灌注期间Brg1的恢复可增强HIR期间Nrf2介导的HO-1诱导表达,从而有效提高抗氧化能力以对抗肝细胞损伤。