Department of Gastroenterology, St. Luc University Hospital, Catholic University of Louvain, Brussels, Belgium.
Eur J Clin Invest. 2010 Jul;40(7):575-84. doi: 10.1111/j.1365-2362.2010.02295.x.
BACKGROUND/AIMS: Conflicting observations exist concerning the role of nuclear factor kappa B (NFjB) in alcoholic liver disease (ALD) in animal models. To date no studies have examined this aspect in human liver tissue. We here assessed cytokines and toll-like receptors (TLRs) expressions in conjunction with NFkappaB activation in non-active end-stage human ALD compared with normal livers and hepatitis C virus (HCV) related end-stage disease.
mRNA and protein expression were examined by quantitative PCR and Western blotting, DNA-binding by electrophoretic mobility shift assays and NFkappaB sub-cellular localization by immunofluorescent staining of livers.
NFkappaB mRNA and protein expression as well as strong DNA-binding were preserved in ALD but significantly down-regulated in HCV compared with normal livers. P50 immunofluorescence was found in hepatocytes and bile ducts in ALD and normal livers, whereas a shift was observed in p65 staining from non-parenchymal cells in normal livers to hepatocytes in ALD. NFkappaB responsive genes mRNA levels IkBalpha and interleukin 6 were significantly higher in ALD compared with HCV. Tumour necrosis factor alpha (TNFalpha), TLRs 3 and 7 mRNA were up-regulated in ALD and HCV compared with normal liver with TNFalpha and TLR7 being the highest in HCV. Strong induction of interferon beta was found in HCV but not in ALD or normal liver tissue.
Persistent NFkappaB activation together with high pro-inflammatory cytokine expression and upregulation of TLR3 and TLR7 is associated with end-stage ALD in humans and could contribute to disease progression even in absence of alcohol intake.
背景/目的：在动物模型中，核因子 kappa B（NFjB）在酒精性肝病（ALD）中的作用存在相互矛盾的观点。迄今为止，尚无研究在人类肝组织中对此进行研究。我们在此评估了非活动终末期人类 ALD 与正常肝脏和丙型肝炎病毒（HCV）相关终末期疾病相比，细胞因子和 Toll 样受体（TLRs）的表达以及 NFkappaB 激活情况。
通过定量 PCR 和 Western blot 检测 mRNA 和蛋白表达，通过电泳迁移率变动分析检测 DNA 结合，通过免疫荧光染色检测 NFkappaB 亚细胞定位。
NFkappaB mRNA 和蛋白表达以及强 DNA 结合在 ALD 中得到保留，但与正常肝脏相比，在 HCV 中明显下调。在 ALD 和正常肝脏中均发现 P50 免疫荧光在肝细胞和胆管中，而在正常肝脏中非实质细胞中的 p65 染色则发生了转移，在 ALD 中转移到了肝细胞中。与 HCV 相比，NFkappaB 反应基因 mRNA 水平 IkBalpha 和白细胞介素 6 在 ALD 中显着升高。TNFalpha、TLRs 3 和 7 在 ALD 和 HCV 中均上调，与正常肝脏相比，TNFalpha 和 TLR7 最高。在 HCV 中发现干扰素 beta 强烈诱导，但在 ALD 或正常肝组织中则没有。
持续的 NFkappaB 激活以及高促炎细胞因子表达和 TLR3 和 TLR7 的上调与人类终末期 ALD 相关，即使在没有饮酒的情况下，也可能导致疾病进展。
