van de Bovenkamp M, Groothuis G M M, Meijer D K F, Olinga P
Department of Pharmacokinetics and Drug Delivery, Groningen University Institute for Drug Exploration, University of Groningen, A. Deusinglaan 1, 9713 AV Groningen, The Netherlands.
Toxicol In Vitro. 2008 Apr;22(3):771-8. doi: 10.1016/j.tiv.2007.11.019. Epub 2007 Dec 8.
Cell culture models have contributed significantly to the study of liver fibrosis, but cannot accurately incorporate in vivo cell-cell and cell-extracellular matrix interactions or account for the heterogeneity of the fibrogenic cell population involved in fibrosis development. Thus, there persists a need for an in vitro model that mimics the in vivo situation more closely, which may be provided by using precision-cut liver slices. In the present study we evaluated human liver slices as a tool to study fibrogenesis and test anti-fibrotic drugs. In this study we examined the responses of fibrogenic cells in human liver slices during control incubation and studied the effect of the anti-fibrotic compound pentoxifylline both during control incubation and after induction of early hepatic stellate cell (HSC) activation by carbon tetrachloride. After prolonged (>24 h) incubation, alphaSMA and pro-collagen 1a1 mRNA expression in human liver slices started to increase. Analysis of synaptophysin and fibulin-2 mRNA expression indicated that both activated HSC and other (myo)fibroblasts may be involved in this process. This response of fibrogenic cells to prolonged incubation of the liver slices was accompanied by an increased collagen protein content and could be inhibited by pentoxifylline. Early HSC activation, which was reflected by increased HSP47 and alphaB-crystallin mRNA expression, was not inhibited by pentoxifylline. Preparation and/or culturing of human liver slices induces fibrogenesis, which may be mediated by both activated HSC and resident liver (myo)fibroblasts and may represent a simple and rapid method to test the effects of potential anti-fibrotic drugs on fibrogenic cells in human liver.
细胞培养模型对肝纤维化的研究做出了重大贡献，但无法准确纳入体内细胞间和细胞与细胞外基质的相互作用，也无法解释参与纤维化发展的致纤维化细胞群体的异质性。因此，仍然需要一种能更紧密模拟体内情况的体外模型，使用精密肝切片可能会提供这样的模型。在本研究中，我们评估了人肝切片作为研究纤维化发生和测试抗纤维化药物的工具。在这项研究中，我们检测了人肝切片在对照孵育期间致纤维化细胞的反应，并研究了抗纤维化化合物己酮可可碱在对照孵育期间以及在四氯化碳诱导早期肝星状细胞（HSC）活化后的作用。长时间（>24小时）孵育后，人肝切片中αSMA和前胶原1a1 mRNA表达开始增加。对突触素和纤连蛋白-2 mRNA表达的分析表明，活化的HSC和其他（肌）成纤维细胞可能都参与了这一过程。肝切片长时间孵育后致纤维化细胞的这种反应伴随着胶原蛋白含量的增加，并且可以被己酮可可碱抑制。早期HSC活化表现为HSP47和αB-晶状体蛋白mRNA表达增加，未被己酮可可碱抑制。人肝切片的制备和/或培养会诱导纤维化，这可能由活化的HSC和驻留肝（肌）成纤维细胞介导，并且可能代表一种简单快速的方法来测试潜在抗纤维化药物对人肝中致纤维化细胞的作用。
