Removal of heparan sulfate by heparinase treatment inhibits FGF-2-dependent smooth muscle cell proliferation in injured rat carotid arteries
Removal of heparan sulfate by heparinase treatment inhibits FGF-2-dependent smooth muscle cell proliferation in injured rat carotid arteries
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DOI：
10.1016/j.atherosclerosis.2004.01.045
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发表时间：
2004-07-01
期刊：
影响因子：
5.3
通讯作者：
Wight, TN
中科院分区：
文献类型：
--
作者：
Kinsella, MG;Irvin, C;Wight, TN
在新生内膜形成过程中，大鼠颈动脉中膜平滑肌细胞（SMC）增生和迁移。在损伤部位释放的成纤维细胞生长因子（FGF-2）与硫酸肝素蛋白多糖（HSPGs）的相互作用对于诱导信号传导是必要的，这引发了动脉平滑肌细胞对fgf依赖性的有丝分裂反应，也作为生长因子在细胞外基质内储存的机制。然而，这些相互作用在新生内膜形成过程中是否至关重要还没有直接测试。本研究采用球囊损伤大鼠颈动脉fgf -2依赖性内侧SMC有丝分裂反应模型，检测血管壁硫酸肝素降解对随后SMC增殖的影响。通过免疫过氧化物酶染色，用软骨素ABC裂解酶和/或肝素裂解酶处理球囊导管大鼠颈动脉可消除血管壁中的硫酸肝素。与此相反，大核蛋白在颈动脉血管壁的分布。结合FGF-2的主要血管壁HSPG没有减少。通过测量损伤后48小时含有5-溴-2'-脱氧尿苷（BrdU）的SMC核的百分比，原位糖胺聚糖消化对损伤后大量注射FGF-2后内侧SMC增殖的影响。损伤后48小时，酶去除硫酸肝素可使BrdU掺入内侧SMC减少60-70% （P < 0.001）。此外，在注射前将FGF-2与肝素预先孵育，可使损伤血管的SMC复制恢复到仅用缓冲液处理的水平（P < 0.01）。这些实验表明，内源性HSPGs对于促进损伤后fgf -2驱动的内侧SMC增殖至关重要，肝素酶治疗可以消除体内fgf -2依赖性反应。2004爱思唯尔爱尔兰有限公司版权所有。
Smooth muscle cells (SMC) of the rat carotid arterial media proliferate and migrate in response to injury during the formation of a neointima. The interaction of fibroblast growth factor (FGF-2), which is released at the site of injury, with heparan sulfate proteoglycans (HSPGs) is necessary to induce signaling, which elicits an FGF-dependent mitogenic response by arterial smooth muscle cells, and also serves as a mechanism for storage of the growth factor within the extracellular matrix. However, whether these interactions are critical during neointimal formation has not been directly tested. In this study, a model of FGF-2-dependent medial SMC mitogenic response in balloon-injured rat carotid artery was used to test the effect of degradation of vessel wall heparan sulfate on subsequent SMC proliferation. Treatment of balloon-catheterized rat carotid arteries with chondroitin ABC lyase and/or heparin lyases eliminated heparan sulfates in the vessel wall, as determined by immunoperoxidase staining. In contrast, the distribution in the carotid vessel wall of the large core protein of perlecan. a major vessel wall HSPG that binds FGF-2, is not decreased. The effect of glycosaminoglycan digestion in situ on medial SMC proliferation in response to a bolus injection of FGF-2 after injury was determined by measuring the percentage of SMC nuclei that incorporated 5-bromo-2'-deoxyuridine (BrdU) 48 h after injury. Enzymatic removal of heparan sulfate reduced BrdU incorporation into medial SMC by 60-70% (P < 0.001) at 48 h after injury. Moreover, pre-incubation of FGF-2 with heparin prior to injection restored SMC replication to the levels present in injured vessels treated with buffer alone (P < 0.01). These experiments indicate that endogenous HSPGs are essential to promote FGF-2-driven medial SMC proliferation following injury, and that heparinase treatment can abrogate FGF-2-dependent responses in vivo. (C) 2004 Elsevier Ireland Ltd. All rights reserved.