Beaulieu A D, Lang F, Belles-Isles M, Poubelle P
Department of Medicine, Université Laval, Québec, Canada.
J Rheumatol. 1987 Aug;14(4):656-61.
We have investigated protein synthesis and release by polymorphonuclear leukocytes (PMN) to compare the protein biosynthetic activity of peripheral blood PMN and inflammatory synovial fluid (SF) PMN from patients with inflammatory arthropathies. We analyzed and compared the protein profiles produced by these cells, using patient matched peripheral blood and SF PMN as well as peripheral blood PMN from normals. Twenty-five patients with either rheumatoid arthritis, psoriatic arthritis or gout were studied. Fluorographs of SDS-polyacrylamide slab gels, performed using cell supernatants from metabolically labelled cells, revealed an increased release of de novo synthesized proteins by inflammatory SF PMN compared to peripheral blood PMN. Under reducing conditions, 4 clearly distinguishable high molecular mass products were observed (Mr 230,000, 185,000, 170,000 and 95,000). Two of the protein bands were found to be gelatin binding (Mr 230,000 and Mr 95,000). By Western blot, the Mr 230,000 protein was found to be fibronectin and the Mr 95,000 protein was shown to be identical to a recently described gelatinase. Thus, the activation of PMN in inflammation is accompanied by an increased release of a number of de novo synthesized proteins, including fibronectin. Our studies directly pertain to the in vivo inflammatory process since the PMN were not activated artificially in vitro.
我们研究了多形核白细胞（PMN）的蛋白质合成与释放，以比较炎症性关节病患者外周血PMN和炎性滑液（SF）PMN的蛋白质生物合成活性。我们分析并比较了这些细胞产生的蛋白质谱，使用与患者匹配的外周血和SF PMN以及正常人的外周血PMN。对25例类风湿关节炎、银屑病关节炎或痛风患者进行了研究。使用代谢标记细胞的细胞上清液进行SDS-聚丙烯酰胺平板凝胶荧光显影，结果显示与外周血PMN相比，炎性SF PMN中从头合成蛋白质的释放增加。在还原条件下，观察到4种明显可区分的高分子量产物（分子量分别为230,000、185,000、170,000和95,000）。发现其中两条蛋白带能结合明胶（分子量为230,000和95,000）。通过蛋白质印迹法，发现分子量为230,000的蛋白质是纤连蛋白，分子量为95,000的蛋白质与最近描述的一种明胶酶相同。因此，炎症中PMN的激活伴随着包括纤连蛋白在内的多种从头合成蛋白质释放的增加。我们的研究直接涉及体内炎症过程，因为PMN并非在体外人工激活的。
