Olerud J E, Chiu D S, Usui M L, Gibran N S, Ansel J C
Department of Medicine, University of Washington, Seattle, USA.
J Invest Dermatol. 1998 Oct;111(4):565-72. doi: 10.1046/j.1523-1747.1998.00330.x.
In a study initially designed to evaluate reinnervation of human cutaneous wounds using an antibody to the neuroneal marker protein gene product (PGP) 9.5, we observed marked immunostaining of cells with morphologic features of fibroblasts in the wounds. PGP 9.5 has recently been shown to be an important enzyme in the highly conserved ubiquitin system of proteolysis. Because the ubiquitin system is known to play an important role in regulating the cell cycle, the presence of PGP 9.5 in cells at a wound site was of considerable interest. Our objectives were to clarify the time frame for the appearance of PGP 9.5 and ubiquitin in wounds, to verify that PGP 9.5 is produced by wound fibroblasts, and to evaluate a potential role for these proteins in the tissue repair process. Standard incisional human wounds were stained with antibodies specific for PGP 9.5 and ubiquitin. At 7 d, stellate cells with morphologic features of fibroblasts stained for PGP 9.5, whereas earlier wounds were generally negative. In 14 and 21 d incised wounds and in chronic granulation tissue from nonhealing ulcers there was strong cellular staining for PGP 9.5 and for ubiquitin. These stellate cells also showed expression of mRNA for PGP 9.5 by reverse transcriptase-polymerase chain reaction in situ hybridization. PGP 9.5 was detected in cultured fibroblasts both by reverse transcriptase-polymerase chain reaction and by northern blot analysis. Confocal microscopy showed colocalization of antibodies to PGP 9.5 and prolyl-4-hydroxylase (a fibroblast marker) as well as colocalization of PGP 9.5 and the platelet derived growth factor beta receptor. We conclude that ubiquitin and PGP 9.5 were expressed by fibroblasts during the granulation tissue and remodeling phases wound healing. The mRNA for PGP 9.5 was demonstrated in stellate cells in chronic wounds and in fibroblasts in culture. The appearance of these degradative proteins in later wounds suggests a downregulation function in the wound healing response.
在一项最初旨在使用针对神经元标记蛋白基因产物（PGP）9.5的抗体评估人类皮肤伤口再神经化的研究中，我们观察到伤口中具有成纤维细胞形态特征的细胞有明显的免疫染色。最近发现PGP 9.5是高度保守的泛素蛋白水解系统中的一种重要酶。由于已知泛素系统在调节细胞周期中起重要作用，伤口部位细胞中PGP 9.5的存在备受关注。我们的目标是明确伤口中PGP 9.5和泛素出现的时间框架，验证PGP 9.5是由伤口成纤维细胞产生的，并评估这些蛋白质在组织修复过程中的潜在作用。标准的人类切开伤口用针对PGP 9.5和泛素的特异性抗体进行染色。在第7天，具有成纤维细胞形态特征的星状细胞对PGP 9.5染色，而早期伤口通常为阴性。在切开14天和21天的伤口以及不愈合溃疡的慢性肉芽组织中，PGP 9.5和泛素均有强烈的细胞染色。这些星状细胞通过逆转录聚合酶链反应原位杂交也显示出PGP 9.5的mRNA表达。通过逆转录聚合酶链反应和Northern印迹分析在培养的成纤维细胞中检测到了PGP 9.5。共聚焦显微镜显示PGP 9.5抗体与脯氨酰-4-羟化酶（一种成纤维细胞标记物）共定位，以及PGP 9.5与血小板衍生生长因子β受体共定位。我们得出结论，在肉芽组织形成和伤口愈合的重塑阶段，成纤维细胞表达泛素和PGP 9.5。在慢性伤口的星状细胞和培养的成纤维细胞中证实了PGP 9.5的mRNA。这些降解蛋白在后期伤口中的出现表明在伤口愈合反应中具有下调功能。
