Connective tissue growth factor expression and action in human corneal fibroblast cultures and rat corneas after photorefractive keratectomy
Connective tissue growth factor expression and action in human corneal fibroblast cultures and rat corneas after photorefractive keratectomy
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DOI：
10.1167/iovs.02-0860
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发表时间：
2003-05-01
影响因子：
4.4
通讯作者：
Schultz, GS
中科院分区：
文献类型：
--
作者：
Blalock, TD;Duncan, MR;Schultz, GS
目的.结缔组织生长因子（CTGF）与几种组织中的纤维化有关。在这项研究中，CTGF和转化生长因子（TGF）-β之间的相互作用进行了评估在人角膜成纤维细胞，和CTGF的蛋白和mRNA的水平和位置进行了测量在大鼠角膜准分子激光消融伤口愈合过程中。将人角膜成纤维细胞与TGF-β 1、TGF-β 2和TGF-β 3亚型孵育，并测量CTGF mRNA和蛋白。CTGF通过使用特异性抗体在培养的成纤维细胞中进行免疫定位。在人角膜成纤维细胞中用中和抗体和CTGF的反义寡核苷酸评估TGF-β和CTGF对胶原合成的调节。CTGF mRNA和蛋白质在大鼠角膜中测量到21天后，准分子切削角膜。CTGF蛋白免疫定位于大鼠角膜屈光性角膜切除术（PRK）后，并建立CTGF mRNA和蛋白在离体大鼠角膜刮片中的存在。所有三种TGF-β亚型刺激CTGF在人角膜成纤维细胞中的表达，并且CTGF在细胞中免疫定位。TGF-β和CTGF均增加角膜成纤维细胞中的胶原合成。此外，CTGF抗体或反义寡核苷酸阻断TGF-β刺激的胶原合成。CTGF蛋白和mRNA在大鼠角膜中增加，直到角膜移植后21天。CTGF的表达也检测到离体大鼠角膜刮片。这些数据表明，角膜细胞在TGF-β刺激后表达CTGF，角膜伤口愈合期间CTGF表达显著增加，并且CTGF介导TGF-β诱导角膜成纤维细胞合成胶原的作用。这些数据支持CTGF促进角膜瘢痕形成的假设，并暗示调节CTGF合成和作用可能是减少角膜瘢痕形成的重要目标。
PURPOSE. Connective tissue growth factor (CTGF) has been linked to fibrosis in several tissues. In this study, the interactions between CTGF and transforming growth factor (TGF)-beta were assessed in human corneal fibroblasts, and the levels and location of CTGF protein and mRNA were measured during healing of excimer laser ablation wounds in rat corneas.METHODS. Human corneal fibroblasts were incubated with TGF-beta1, -beta2, and -beta3 isoforms, and CTGF mRNA and protein were measured. CTGF was immunolocalized in the cultured fibroblasts by using, a specific antibody. Regulation of Collagen synthesis by TGF-beta and CTGF was assessed in human corneal fibroblasts with a neutralizing antibody and an antisense oligonucleotide to CTGF. CTGF mRNA and protein were measured in rat corneas up to day 21 after excimer ablation of the cornea. CTGF protein was immunolocalized in rat corneas after photorefractive keratectomy (PRK), and the presence of CTGF mRNA and protein in ex vivo rat corneal scrapings was established.RESULTS. All three TGF-beta isoforms stimulated expression of CTGF in human corneal fibroblasts, and CTGF was immunolocalized in the cells. Both TGF-beta and CTGF increased Collagen synthesis in corneal fibroblasts. Furthermore, CTGF antibody or antisense oligonucleotide blocked TGF-beta-stimulated collagen synthesis. CTGF protein and mRNA increased in rat corneas through day 21 after PRK. CTGF expression was also detected in ex vivo scrapings of rat corneas.CONCLUSIONS. These data demonstrate that CTGF is expressed by corneal cells after stimulation by TGF-beta, that CTGF expression increases significantly during corneal wound healing, and that CTGF mediates the effects of TGF-beta induction of Collagen synthesis by corneal fibroblasts. These data support the hypothesis that CTGF promotes corneal scar formation and imply that regulating CTGF synthesis and action may be an important goal for reducing corneal scarring.