Advanced glycation end products induce production of reactive oxygen species via the activation of NADPH oxidase in murine hepatic stellate cells
Advanced glycation end products induce production of reactive oxygen species via the activation of NADPH oxidase in murine hepatic stellate cells
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DOI：
10.1016/j.jhep.2009.12.007
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发表时间：
2010-03-01
影响因子：
25.7
通讯作者：
van Grunsven, Leo A.
中科院分区：
文献类型：
--
作者：
Guimaraes, Eduardo L. M.;Empsen, Christophe;van Grunsven, Leo A.
背景与目的：已知晚期糖基化终产物在代谢综合征中起重要作用，最近被认为有助于肝纤维化的发展。然而，对于晚期糖基化终产物对肝星状细胞的影响知之甚少，肝星状细胞是肝纤维化发展的主要贡献者。因此，我们研究了晚期糖基化终产物对活性氧生成的影响，活性氧是激活肝星状细胞的主要特征。方法：用不同底物培养牛血清白蛋白生成三种不同类型的晚期糖基化终产物。采用RTq-PCR、免疫荧光和western blotting检测晚期糖基化终产物受体的存在。DCFH-DA法测定活性氧生成。结果：肝星状细胞表达5种晚期糖基化终产物受体：半乳糖凝集素-3、CD36、SR-Al、SR-BI和RAGE。除SR-BI外，所有受体在HSC活化过程中均呈上调。所有三种晚期糖基化终产物类型均诱导活性氧生成。DPI和NSC分别是NADPH氧化酶和Rac1抑制剂，它们抑制活性氧的产生。Rottlerin，一种常被用作PKC δ抑制剂的分子，也废除了活性氧的产生。SiRNA介导的p47（phox）、Rac1和PKC δ的敲低降低了晚期糖基化终产物诱导的活性氧的产生，从而确定了这些蛋白在活性氧诱导中的作用。结论：晚期糖基化终产物在肝星状细胞中诱导活性氧生成的证明揭示了晚期糖基化终产物诱导代谢综合征肝纤维化的潜在新途径。(C) 2010年欧洲肝脏研究协会。Elsevier B.V.版权所有。
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.Methods: 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.Results: Hepatic stellate cells express five advanced glycation end product receptors: Galectin-3, CD36, SR-Al, 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 PKC delta inhibitor, also abrogated reactive oxygen species production. SiRNA mediated knockdown of p47(phox), Rac1 and PKC delta decreased reactive oxygen species production induced by advanced glycation end products, establishing a role for these proteins in reactive oxygen species induction.Conclusions: 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. (C) 2010 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.