The role of AKT1 and autophagy in the protective effect of hydrogen sulphide against hepatic ischemia/reperfusion injury in mice
The role of AKT1 and autophagy in the protective effect of hydrogen sulphide against hepatic ischemia/reperfusion injury in mice
复制标题
AKT1和自噬在硫化氢对小鼠肝缺血/再灌注损伤的保护作用中的作用
DOI：
10.4161/auto.19927
复制
发表时间：
2012-06-01
期刊：
影响因子：
13.3
通讯作者：
Jiang, Hongchi
中科院分区：
文献类型：
--
作者：
Wang, Dawei;Ma, Yong;Jiang, Hongchi
硫化氢（H2S）对肝缺血再灌注（I/R）损伤具有保护作用。然而，H2S作用的确切机制在很大程度上仍然未知。本研究旨在探讨PtdIns3K-AKT1通路和自噬在H2S对肝I/R损伤的保护作用中的作用。原代培养的小鼠肝细胞和NaHS （H2S供体）预处理的肝脏分别暴露于缺氧/再氧化（a /R）和I/R。在某些组中，还单独或同时使用LY294002 （akt1特异性抑制剂）、3-甲基腺嘌呤（3MA，自噬抑制剂）或雷帕霉素（自噬增强剂）进行预处理。检测细胞活力、P-AKT1、T-AKT1、LC3和BECN1的表达。通过血清转氨酶、炎性细胞因子、细胞凋亡及组织学检查判断肝损伤程度。采用GFP-LC3再分布和透射电镜检测自噬活性。H2S预处理激活肝细胞PtdIns3K-AKT1信号。LY294002可抑制AKT1的激活，减弱H2S对肝细胞A/R和肝I/R损伤的保护作用。H2S在体外和体内均能抑制肝自噬。3MA进一步降低自噬也降低了H2S的保护作用，而雷帕霉素可以逆转自噬抑制作用，增强H2S对肝细胞A/R和肝I/R损伤的保护作用。综上所述，H2S至少部分通过AKT1激活而不是自噬来保护肝细胞A/R和肝I/R损伤。自噬激动剂可以通过逆转H2S对自噬的抑制来增强这种肝保护作用。
Hydrogen sulphide (H2S) exerts a protective effect in hepatic ischemia-reperfusion (I/R) injury. However, the exact mechanism of H2S action remains largely unknown. This study was designed to investigate the role of the PtdIns3K-AKT1 pathways and autophagy in the protective effect of H2S against hepatic I/R injury. Primary cultured mouse hepatocytes and livers with or without NaHS (a donor of H2S) preconditioning were exposed to anoxia/reoxygenation (A/R) and I/R, respectively. In certain groups, they were also pretreated with LY294002 (AKT1-specific inhibitor), 3-methyladenine (3MA, autophagy inhibitor) or rapamycin (autophagy enhancer), alone or simultaneously. Cell viability, expression of P-AKT1, T-AKT1, LC3 and BECN1 were examined. The severity of liver injury was measured by the levels of serum aminotransferase and inflammatory cytokine, apoptosis and histological examination. GFP-LC3 redistribution and transmission electron microscopy were used to test the activity of autophagy. H2S preconditioning activated PtdIns3K-AKT1 signaling in hepatocytes. LY294002 could abolish the AKT1 activation and attenuate the protective effect of H2S on hepatocytes A/R and hepatic I/R injuries. H2S suppressed hepatic autophagy in vitro and in vivo. Further reducing autophagy by 3MA also diminished the protective effect of H2S, while rapamycin could reverse the autophagy inhibitory effect and enhance the protective effect of H2S against hepatocytes A/R and hepatic I/R injuries, consequently. Taken together, H2S protects against hepatocytic A/R and hepatic I/R injuries, at least in part, through AKT1 activation but not autophagy. An autophagy agonist could be applied to potentiate this hepatoprotective effect by reversing the autophagy inhibition of H2S.