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首页> 外文期刊>Investigative ophthalmology & visual science >A Connective Tissue Growth Factor Signaling Receptor in Corneal Fibroblasts
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A Connective Tissue Growth Factor Signaling Receptor in Corneal Fibroblasts

机译：角膜成纤维细胞中的结缔组织生长因子信号受体

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Purpose.: To biochemically characterize the receptor for connective tissue growth factor (CTGF) of human corneal fibroblasts (HCF). Methods.: Radiolabeled recombinant human CTGF was used to determine the specificity and time course of binding to low-passage cultures of HCF. The affinity and number of receptors present were calculated by Scatchard and best-fit analyses. In vitro immunoprecipitation assays with radiolabeled CTGF and soluble mannose 6-phosphate/insulin-like growth factor 2 receptor (M6P/IGF-2-R) alone, or with CTGF-related growth factors were conducted. Additionally, 125I-CTGF-binding and CTGF-stimulated proliferation were measured in cultures of M6P/IGF-2-R knockout fibroblasts. Results.: Binding of 125I-CTGF to fibroblast cultures was significantly displaced by CTGF, but not by related growth factors. Scatchard plot analysis indicated the presence of both a high-affinity, low-abundance binding site, and a low-affinity, high-abundance binding site; whereas, the best-fit analysis suggests a single high-affinity, low-abundance binding site. A 280 kDa complex containing cross-linked 125I-CTGF was immunoprecipitated by antibodies to CTGF or M6P/IGF-2-R. M6P/IGF-2-R knockout cells have a reduced proliferative response to TGF-?2, and don't proliferate at all in response to CTGF. Conclusions.: CTGF binds to the M6P/IGF-2-R with high affinity, and the M6P/IGF-2-R is required for CTGF-stimulated proliferation in fibroblasts. These observations suggest that the M6P/IGF-2-R may be a new antifibrotic target.
机译：目的：生化表征人类角膜成纤维细胞（HCF）的结缔组织生长因子（CTGF）受体。方法：使用放射性标记的重组人CTGF来确定与HCF低传代培养物结合的特异性和时程。通过Scatchard和最佳拟合分析计算存在的受体的亲和力和数量。进行了仅用放射性标记的CTGF和可溶性甘露糖6-磷酸/胰岛素样生长因子2受体（M6P / IGF-2-R）或与CTGF相关的生长因子的体外免疫沉淀测定。另外，在M6P / IGF-2-R敲除成纤维细胞的培养物中测量了125I-CTGF结合和CTGF刺激的增殖。结果：125 I-CTGF与成纤维细胞培养物的结合被CTGF显着取代，但未被相关的生长因子取代。斯卡查德图分析表明存在高亲和力，低丰度的结合位点和低亲和力，高丰度的结合位点。而最佳拟合分析表明单个高亲和力，低丰度的结合位点。含有交联的125I-CTGF的280 kDa复合物被CTGF或M6P / IGF-2-R抗体免疫沉淀。 M6P / IGF-2-R敲除细胞对TGF-β2的增殖反应降低，并且根本不对CTGF增殖。结论：CTGF以高亲和力与M6P / IGF-2-R结合，而CTGF刺激的成纤维细胞增殖需要M6P / IGF-2-R。这些观察结果表明，M6P / IGF-2-R可能是新的抗纤维化靶标。

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