Department of Cell Biology, Vrije Universiteit, Laarbeeklaan 103, Brussels, Belgium.
J Hepatol. 2010 Mar;52(3):389-97. doi: 10.1016/j.jhep.2009.12.007. Epub 2009 Dec 24.
BACKGROUND & AIMS: Advanced glycation end products are known to play an important role in the metabolic syndrome and were recently suggested to contribute to liver fibrosis development. However, little is known about the effect of advanced glycation end products on hepatic stellate cells, the major contributors to liver fibrosis development. We therefore studied the effect of advanced glycation end products on reactive oxygen species generation, a main feature for the activation hepatic stellate cells.
Three different types of advanced glycation end products were generated by BSA incubation with different substrates. The presence of advanced glycation end product receptors was examined by RTq-PCR, immunofluorescence and western blotting. Reactive oxygen species production was measured using DCFH-DA.
Hepatic stellate cells express five advanced glycation end product receptors: Galectin-3, CD36, SR-AI, SR-BI and RAGE. All receptors, except SR-BI, showed up-regulation during HSC activation. All three advanced glycation end product types induced reactive oxygen species generation. DPI and NSC, a NADPH oxidase and a Rac1 inhibitor respectively, inhibited reactive oxygen species production. Rottlerin, a molecule often used as a PKCdelta inhibitor, also abrogated reactive oxygen species production. SiRNA mediated knockdown of p47(phox), Rac1 and PKCdelta decreased reactive oxygen species production induced by advanced glycation end products, establishing a role for these proteins in reactive oxygen species induction.
The demonstration of advanced glycation end product-induced reactive oxygen species generation in hepatic stellate cells unveils a potential new route through which advanced glycation end products induce liver fibrosis in the metabolic syndrome.
已知糖基化终产物在代谢综合征中起着重要作用，最近有研究表明其有助于肝纤维化的发展。然而，关于糖基化终产物对肝星状细胞（肝纤维化发展的主要贡献者）的影响知之甚少。因此，我们研究了糖基化终产物对活性氧（ROS）生成的影响，ROS 是肝星状细胞激活的主要特征之一。
通过 BSA 与不同底物孵育生成三种不同类型的糖基化终产物。通过 RTq-PCR、免疫荧光和 Western blot 检测糖基化终产物受体的存在。使用 DCFH-DA 测量 ROS 生成。
肝星状细胞表达五种糖基化终产物受体：半乳糖凝集素-3、CD36、SR-AI、SR-BI 和 RAGE。除了 SR-BI 之外，所有受体在 HSC 激活期间均上调。三种糖基化终产物类型均诱导 ROS 生成。DPI 和 NSC，分别是 NADPH 氧化酶和 Rac1 抑制剂，抑制 ROS 生成。作为 PKCδ抑制剂常用的分子罗特林也阻断了 ROS 的生成。p47(phox)、Rac1 和 PKCδ 的 siRNA 介导的敲低减少了糖基化终产物诱导的 ROS 生成，证实了这些蛋白在 ROS 诱导中的作用。
证明糖基化终产物诱导肝星状细胞中 ROS 的生成揭示了糖基化终产物在代谢综合征中诱导肝纤维化的新途径。
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