Borkham-Kamphorst Erawan, Alexi Pascal, Tihaa Lidia, Haas Ute, Weiskirchen Ralf
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, RWTH Aachen University Hospital, Germany.
Institute of Molecular Pathobiochemistry, Experimental Gene Therapy and Clinical Chemistry, RWTH Aachen University Hospital, Germany.
Biochem Biophys Res Commun. 2015 Feb 13;457(3):307-13. doi: 10.1016/j.bbrc.2014.12.106. Epub 2015 Jan 7.
Platelet-derived growth factor-D (PDGF-D) is a more recent recognized growth factor involved in the regulation of several cellular processes, including cell proliferation, transformation, invasion, and angiogenesis by binding to and activating its cognate receptor PDGFR-β. After bile duct ligation or in the carbon tetrachloride-induced hepatic fibrosis model, PDGF-D showed upregulation comparable to PDGF-B. Moreover, adenoviral PDGF-D gene transfer induced hepatic stellate cell proliferation and liver fibrosis. We here investigated the molecular mechanism of PDGF-D involvement in liver fibrogenesis. Therefore, the GRX mouse cell line was stimulated with PDGF-D and evaluated for fibrotic markers and PDGF-D signaling pathways in comparison to the other PDGF isoforms. We found that PDGF-D failed to enhance Col I and α-smooth muscle actin (α-SMA) production but has capacity to upregulate expression of the tissue inhibitor of metalloprotease 1 (TIMP-1) resulting in attenuation of MMP-2 and MMP-9 gelatinase activity as indicated by gelatinase zymography. This phenomenon was restored through application of a PDGF-D neutralizing antibody. Unexpectedly, PDGF-D incubation decreased both PDGFR-α and -β in mRNA and protein levels, and PDGF-D phosphorylated typrosines specific for PDGFR-α and -β. We conclude that PDGF-D intensifies fibrogenesis by interfering with the fibrolytic activity of the TIMP-1/MMP system and that PDGF-D signaling is mediated through both PDGF-α and -β receptors.
血小板衍生生长因子-D(PDGF-D)是一种最近才被认识的生长因子,它通过与同源受体PDGFR-β结合并激活该受体,参与多种细胞过程的调节,包括细胞增殖、转化、侵袭和血管生成。在胆管结扎或四氯化碳诱导的肝纤维化模型中,PDGF-D的上调程度与PDGF-B相当。此外,腺病毒介导的PDGF-D基因转移可诱导肝星状细胞增殖和肝纤维化。我们在此研究了PDGF-D参与肝纤维化形成的分子机制。因此,用PDGF-D刺激GRX小鼠细胞系,并与其他PDGF异构体相比,评估其纤维化标志物和PDGF-D信号通路。我们发现,PDGF-D未能增强I型胶原和α-平滑肌肌动蛋白(α-SMA)的产生,但有能力上调金属蛋白酶组织抑制剂1(TIMP-1)的表达,导致MMP-2和MMP-9明胶酶活性减弱,这通过明胶酶谱法得以证实。通过应用PDGF-D中和抗体,这种现象得以恢复。出乎意料的是,PDGF-D孵育降低了PDGFR-α和-β的mRNA和蛋白水平,以及PDGF-D对PDGFR-α和-β特异的酪氨酸磷酸化。我们得出结论,PDGF-D通过干扰TIMP-1/MMP系统的纤溶活性来加剧纤维化形成,并且PDGF-D信号传导是通过PDGF-α和-β受体介导的。