Characterization of Tumor Necrosis Factor Alpha-Induced Alteration of Glycosaminoglycans in Cultured Cells : Comparison among Vascular Smooth-Muscle Cells, Vascular Endothelial Cells, Chang Liver Cells and LLC-PK1 Cells

Abstract

We investigated the alteration of glycosaminoglycans (GAGs) induced by recombinant human tumor necrosis factor alpha (rhTNFα) using confluent cultures of bovine aortic smooth-muscle cells, bovine aortic endothelial cells, Chang liver cells and porcine kidney LLC-PK1 cells. It was found that the incorporation of both [³⁵S]sulfate and [³H]glucosamine into GAGs in the trypsinate fraction of the cell layer was significantly decreased by rhTNFα in vascular smooth-muscle cells and vascular endothelial cells; the incorporation of [³⁵S]sulfate was increased but that of [³H]glucosamine was unchanged in Chang liver cells; the incorporation of both [³⁵S]sulfate and [³H]glucosamine was increased by rhTNFα in LLC-PK1 cells. In the conditioned medium, the incorporation of both [³⁵S]sulfate and [³H]glucosamine was not greatly changed by rhTNFα in all tested cell types. Characterization of GAGs revealed that each cell type uniquely altered its GAGs after rhTNFα treatment; the cytokine-induced alteration of each GAG component was not necessarily the same among different cell types. It was therefore concluded that rhTNFα-induced alteration of GAGs is dependent upon cell type.