Glycocalyx on the surface of endothelium has been suggested to be involved in vascular permeability and anticoagulation. In the present study, we demonstrated that fluid laminar shear stress enhanced a glycosaminoglycan (GAG) synthesis in porcine aortic endothelial cells, in vitro. Shear stress (15, 40 dyn/cm2) for 24 hours significantly increased GAG synthesis, assayed by [35S]sulfate incorporation, in "medium" fraction and "trypsinated" fraction which includes GAGs derived from the cell surface and from the solubilized matrix. Increased GAGs in the trypsinated and medium fractions consisted of mainly heparan sulfate and chondroitin/dermatan sulfate, respectively. Both heparan and chondroitin/dermatan sulfate increases are required to expose the cells to shear stress for more than 24 hours. Shear-stress-induced increase in GAG synthesis was concomitant with a decrease in DNA synthesis and an increase in protein synthesis. These data indicate that relatively high shear stress may suppress atherogenesis by changing endothelial GAG synthesis.