Haralson M A, Jacobson H R, Hoover R L
Department of Pathology, Vanderbilt University School of Medicine, Nashville, Tennessee.
Lab Invest. 1987 Nov;57(5):513-23.
Studies have been performed to evaluate both the relative amounts and the molecular forms of the collagens synthesized by cultured rat kidney mesangial (RKM) cells. The collagens secreted into the culture medium and extracted from the cell layers of cultured RKM cells were isolated after limited pepsin digestion and differential salt fractionation. Polyacrylamide gel electrophoresis under denaturing conditions of the RKM cell collagens indicated the presence of components corresponding to the chains present in types I, III, IV, and V collagen. Analysis of each fraction by carboxymethyl-trisacryl chromatography revealed that approximately 95% of the total collagen synthesized by the cultured mesangial cells was type I and that approximately one-half of this genetic type of collagen was present as type I trimers. The type IV molecules synthesized by RKM cells exhibited the molecular structure [alpha 1(IV)]3, whereas the type V molecules had the molecular composition [alpha 1(V)]2 alpha 2(V). Furthermore, in contrast to the type I and V collagens which were identified in both the secreted and cell-associated fractions, no type III or type IV components were detected among the collagens retained by the cells. These data establish the relative proportions of the collagens synthesized by cultured mesangial cells and suggest that the process of cell culture may induce a wound-healing or sclerosing phenotype in the glomerular mesangial cell.
已有研究评估培养的大鼠肾系膜(RKM)细胞合成的胶原蛋白的相对含量和分子形式。在有限的胃蛋白酶消化和分级盐析后,分离出分泌到培养基中以及从培养的RKM细胞层中提取的胶原蛋白。在变性条件下对RKM细胞胶原蛋白进行聚丙烯酰胺凝胶电泳,结果表明存在与I、III、IV和V型胶原蛋白中的链相对应的成分。通过羧甲基三丙烯酸酯色谱法对每个组分进行分析,结果显示培养的系膜细胞合成的总胶原蛋白中约95%为I型,且该遗传类型的胶原蛋白中约一半以I型三聚体形式存在。RKM细胞合成的IV型分子呈现[α1(IV)]3分子结构,而V型分子的分子组成为[α1(V)]2α2(V)。此外,与在分泌组分和细胞相关组分中均鉴定出的I型和V型胶原蛋白不同,在细胞保留的胶原蛋白中未检测到III型或IV型成分。这些数据确定了培养的系膜细胞合成的胶原蛋白的相对比例,并表明细胞培养过程可能诱导肾小球系膜细胞出现伤口愈合或硬化表型。