BETA-1 INTEGRIN-MEDIATED COLLAGEN GEL CONTRACTION IS STIMULATED BY PDGF
BETA-1 INTEGRIN-MEDIATED COLLAGEN GEL CONTRACTION IS STIMULATED BY PDGF
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DOI：
10.1016/0014-4827(90)90305-t
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发表时间：
1990-02-01
影响因子：
3.7
通讯作者：
RUBIN, K
中科院分区：
文献类型：
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作者：
GULLBERG, D;TINGSTROM, A;RUBIN, K
原代大鼠肝细胞和成纤维细胞与I型胶原的附着由非RGD依赖性β 1整联蛋白基质受体介导。在这份报告中，我们描述了一种新的96孔微量滴定板测定成纤维细胞介导的收缩浮动胶原I型凝胶的定量。胎牛血清和血小板衍生生长因子（PDGF），而不是转化生长因子-β 1，刺激原代大鼠心脏成纤维细胞和正常人二倍体成纤维细胞（AG 1518）在24小时温育期内使胶原凝胶收缩至小于初始凝胶体积的10%。针对大鼠肝细胞整联蛋白β 1链的兔多克隆抗体抑制PDGF刺激的胶原凝胶收缩。抗β 1整联蛋白IgG对收缩的抑制活性可以通过加入更高剂量的PDGF来克服。收缩过程不被阻断的抗纤连蛋白IgG，也不是由合成肽含有三肽精氨酸-甘氨酸-天冬氨酸（RGD），在浓度很容易阻止成纤维细胞附着到纤连蛋白包被的平面基板。自体纤连蛋白或含有三肽Arg-Gly-Glu的对照肽没有影响。在胶原凝胶内生长的成纤维细胞上的免疫荧光显微镜显示β 1整联蛋白的点状分布和缺乏可检测水平的内源性产生的纤连蛋白。总的来说，这些数据表明整合素胶原蛋白受体与胶原纤维的亲和力，不同于先前描述的RGD依赖性纤连蛋白受体，在纤连蛋白非依赖性PDGF刺激的胶原蛋白凝胶收缩过程中的作用。
The attachment of primary rat hepatocytes and fibroblasts to collagen type I is mediated by non=RGD-dependent .beta.1 integrin matrix receptors. In this report we describe a novel 96-well microtiter plate assay for the quantification of fibroblast-mediated contraction of floating collagen type I gels. Fetal calf serum and platelet-derived growth factor (PDGF), but not transforming growth factor-.beta.1, stimulated primary rat heart fibroblasts and normal human diploid fibroblasts (AG 1518) to contract collagen gels to less than 10% of the initial gel volume within a 24-h incubation period. Rabbit polyclonal antibodies directed to the rat hepatocyte integrin .beta.1-chain inhibited the PDGF-stimulated collagen gel contraction. The inhibitory activity on contraction of the anti-.beta.1 integrin IgG could be overcome by adding higher doses of PDGF. The contraction process was not blocked by anti-fibronectin IgG nor by synthetic peptides containing the tripeptide Arg-Gly-Asp (RGD), in concentrations that readily blocked fibroblast attachment to fibronectin-coated planar substrates. Autologous fibronectin or control peptides containing the tripeptide Arg-Gly-Glu were without effect. Immunofluorescence microscopy on fibroblasts grown within collagen gels revealed a punctate distribution of the .beta.1 integrin and a lack of detectable levels of endogenously produced fibronectin. Collectively these data suggest a role for integrin collagen receptors with affinity for collagen fibers, distinct from the previously described RGD-dependent fibronectin receptors, in the fibronectin-independent PDGF-stimulated collagen gel contraction process.