AMPK激活剂减轻TNF-α诱导的爆发性肝损伤

AMPK Activator Alleviates TNF-α-induced Fulminant Liver Injury in Mice

探讨腺苷酸活化蛋白激酶(AMP activated protein kinase,AMPK)激活剂A-769662对肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)诱导的爆发性肝损伤中肝细胞凋亡的影响及机制。本研究在雄性BALB/c小鼠经腹腔内注射肿瘤坏死因子α(tumor necrosis factor alpha,TNF-α)及D氨基半乳糖胺(D-galactosamine,D-Gal),诱导爆发性肝炎模型。实验分为4组:正常对照组、A-769662单独处理组、模型组和A-769662干预组。采用比色法检测血浆转氨酶活性及肝组织中caspase-8、caspase-9、caspase-3的相对活性,Western blotting法检测肝组织中激活型caspase-3蛋白水平,TUNEL法检测肝细胞凋亡,HE染色观察肝组织病理变化,并记录各组小鼠生存情况。A-769662干预抑制了TNF-α/D-Gal诱导的肝组织中caspase-8、caspase-9、caspase-3的活性和激活型caspase-3蛋白表达水平的升高,显著减少肝细胞凋亡数目,明显下调血浆中天冬氨酸氨基转氨酶(aspartate transaminase,AST)与丙氨酸氨基转氨酶(alanine transaminase,ALT)水平,减轻肝组织病理学改变,提高小鼠生存率。AMPK激活剂A-769662在TNF-α诱导的爆发性肝损伤中,可发挥抗凋亡保护效应,这可能是其在保肝效应的新机制。

To investigate the potential effects of adenosine 5＇-monophosphate-activated protein kinase （AMPK） activatorA-69662 on tumor necrosis factor alpha （TNF-α）-induced fulminant hepatic injury. TNF-α and D-galactosamine （D-Gal） were intraperitoneally injected in male BALB/c to inducefulminant liver injury in this study. Mice were randomly divided into four groups： nomal control group, A-769662 control gruop, model group and A-769662 intervention group. Plasma transaminase activity and liver caspase-8, caspase-9, caspase-3 relative activity wereanalyzed with Colorimetric kits. The level ofcleaved caspase-3 proteinin liver was detected by West- ern blotting, hepatocyte apoptosis wasdetermined by TUNEL, liver tissue HE staining was used to observe the pathological changes. In addition, the survival rate of the experimental animals was monitored. A-769662 intervention suppressed TNF-α/D-Gal-inducedelevatedactivity of caspase-8, caspase-9, caspase-3, downregnlated the level of cleaved caspase-3 protein in liver tissue, decreased the increasing aspartate transaminase （AST） and alanine transaminase （ALT） levels in plasma, and also significantly reducedthe count of apoptotic hepatocytes and alleviatethe histopatholo-gical abnormalities in liver tissue, improved the survival rate of mice. The AMPK activator A-769662 could provide anti-apoptotic and protective benefits in TNF-α-induced fulminant liver injury, which might be a novel mechanism underlying its hepatoprotective actions.