Stimulation of serglycin and CD44 mRNA expression in endothelial cells exposed to TNF-alpha and IL-1alpha.

Serglycin is a widely distributed proteoglycan, previously assumed to be hematopoietic cell specific. However, the results presented show that serglycin mRNA is expressed outside the hematopoietic cell system. High levels of serglycin mRNA were detected in endothelial cells and smooth muscle cells, whereas low levels were detected in skin fibroblasts. To further analyze the importance of serglycin in endothelial cells, the expression of serglycin mRNA was measured following activation of an endothelial cell line derived from human umbilical cord vein (HUV-EC-C), by the proinflammatory cytokines TNF-alpha and IL-1alpha. The level of serglycin mRNA increased in a time- and dose-dependent way. TNF-alpha (7 ng/ml) was the most potent inducer, increasing the level of serglycin mRNA 2.5 times after 24 h of stimulation. Serglycin has been shown to be a ligand for CD44, a membrane protein expressed in endothelial cells. Following stimulation of the endothelial cells, the level of CD44 mRNA also increased. Again, TNF-alpha (7 ng/ml) turned out to be the most potent inducer, increasing the level of CD44 mRNA 5.5 times after 24 h of stimulation. Both TNF-alpha and IL-1alpha stimulation of the endothelial cells resulted in an increase in the total incorporation of [(35)S]sulfate into macromolecules, which probably indicates an increase in the total production of proteoglycans. A stimulation of endothelial cells by proinflammatory agents resulted in an increase in both serglycin and CD44 mRNA expression, indicating that serglycin, as well as CD44, may participate in the inflammatory process of leukocyte migration.