Persistence of scleroderma-like phenotype in normal fibroblasts after prolonged exposure to soluble mediators from mononuclear cells.
Persistence of scleroderma-like phenotype in normal fibroblasts after prolonged exposure to soluble mediators from mononuclear cells.
复制标题
长期暴露于单核细胞的可溶性介质后，正常成纤维细胞中硬皮病样表型的持续存在。
DOI：
10.1002/art.1780290108
复制
发表时间：
1986
影响因子：
--
通讯作者：
Rodnan,GP
中科院分区：
文献类型：
--
作者：
Worrall,JG;Whiteside,TL;Prince,RK;Buckingham,RB;Stachura,I;Rodnan,GP
单核细胞（MNC-SN）的上清液显示可增加培养的正常皮肤成纤维细胞的糖胺聚糖（GAG）合成。来自进行性系统性硬化症（PSS，硬皮病）患者皮肤的成纤维细胞反应迟钝。我们将从正常成人皮肤外植体中生长出来的成纤维细胞暴露于MNC‐SN培养长达30代。通过将正常MNC与伴刀豆球蛋白A孵育获得MNC-SN。通过胰蛋白酶消化和每7天1：2分离培养物，对4个实验系、4个正常对照系和3个PSS对照系进行传代。在第三和第五次传代时，从MNC-SN中取出部分实验成纤维细胞，然后在单独的培养基中传代。进行细胞计数、GAG测定和电子显微镜检查，并在第3、5和8代时测定短暂再暴露于MNC‐SN后的GAG增加。在正常皮肤成纤维细胞中，通过3 H-葡萄糖胺摄取测量的基线GAG产生较低，并且在重新暴露于MNC-SN后增加多达15倍。相比之下，实验和PSS系的产量都很高，并且再次暴露于MNC-SN后的增加一直很小。这种PSS样行为在第三代和第五代从MNC-SN中取出的实验成纤维细胞中持续存在。实验组和硬皮病组成纤维细胞的生长速度比对照组慢。在超微结构上，硬皮病和实验性真皮成纤维细胞与正常成纤维细胞的不同之处在于它们的椭圆形细胞形状、核膜中的凹陷、许多细胞器和微丝束、突出的高尔基体和核内包涵体。这些实验表明，在体外经受MNC-SN的正常成人真皮成纤维细胞获得了持续多代的硬皮病样表型。
Supernatants of mononuclear cells (MNC‐SN) were shown to increase synthesis of glycosaminoglycan (GAG) by cultured normal dermal fibroblasts. Fibro‐blasts from the skin of patients with progressive systemic sclerosis (PSS, scleroderma) were hyporesponsive. We exposed fibroblasts outgrowing from explants of normal adult skin to MNC‐SN for up to 30 generations in culture. MNC‐SN were obtained by incubating normal MNC with concanavalin A. Four experimental, 4 normal control, and 3 PSS control lines were passaged by trypsinizing and splitting the cultures 1:2 every 7 days. At the third and fifth passages, portions of the experimental fibroblasts were removed from MNC‐SN, then passaged in medium alone. Cell counts, assays for GAG, and electron microscopy were performed and increases in GAG after brief reexposure to MNC‐SN were determined at the third, fifth, and eighth passages. In normal dermal fibroblasts, baseline GAG production, measured by3H‐glucosamine uptake, was low and increased as much as 15 times after reexposure to MNC‐SN. In contrast, production was high in both experimental and PSS lines, and increases after reexposure to MNC‐SN were consistently small. This PSS‐like behavior persisted in experimental fibroblasts removed from MNC‐SN at the third and fifth passages. Growth of experimental and scleroderma fibroblasts was slower than that of control fibroblasts. Ultra‐structurally, both scleroderma and experimental dermal fibroblasts differed from normal fibroblasts by their oval cellular shape, indentations in nuclear membrane, numerous organelles and bundles of microfilaments, prominent Golgi, and intranuclear inclusions. These experiments indicate that normal adult dermal fibroblasts subjected to MNC‐SN in vitro acquire a scleroderma‐like phenotype that persists for many generations.