Maheshwari R K, Kedar V P, Bhartiya D, Coon H C, Kang Y H
Department of Pathology, Uniformed Services University of the Health Sciences, Bethesda, Maryland.
J Biol Regul Homeost Agents. 1990 Jul-Sep;4(3):117-24.
Fibronectin (FN), a normal plasma and extracellular matrix glycoprotein, plays a significant role in various phases of wound healing. At wound site FN is synthesized locally by various cell types involved in the healing process (viz. epithelial, endothelial, fibroblast and macrophage cells) or deposited from the plasma. The present study was undertaken to investigate the in vitro effect of IFN on FN synthesis as well as release in the culture medium by various cell types. Indirect immunofluorescence and immunoelectron microscopy studies, using specific antibodies, revealed that IFN treatment resulted in significantly more staining for FN as compared to untreated control cells. Metabolic labeling with 35S-methionine, immunoprecipitation and SDS-page studies showed an increase in FN synthesis and release by IFN treated cells. In addition, to determine whether this increased synthesis was reflected at mRNA levels, poly (A)+ RNA was isolated from human lung epithelial cells (A549) and probed with FN specific cDNA. We found that IFN treatment increased the level of FN mRNA.
纤连蛋白(FN)是一种正常的血浆和细胞外基质糖蛋白,在伤口愈合的各个阶段发挥着重要作用。在伤口部位,FN由参与愈合过程的各种细胞类型(即上皮细胞、内皮细胞、成纤维细胞和巨噬细胞)在局部合成,或从血浆中沉积。本研究旨在探讨干扰素对各种细胞类型FN合成以及在培养基中释放的体外作用。使用特异性抗体进行的间接免疫荧光和免疫电子显微镜研究表明,与未处理的对照细胞相比,干扰素处理导致FN染色显著增加。用35S-甲硫氨酸进行代谢标记、免疫沉淀和SDS-聚丙烯酰胺凝胶电泳研究表明,干扰素处理的细胞中FN合成和释放增加。此外,为了确定这种增加的合成是否在mRNA水平上得到体现,从人肺上皮细胞(A549)中分离出聚腺苷酸(poly (A)+)RNA,并用FN特异性cDNA进行探针杂交。我们发现,干扰素处理增加了FN mRNA的水平。