Effects of tumor necrosis factor on prostacyclin production and the barrier function of human endothelial cell monolayers

The endothelium controls the influx of macromolecules into the tissues, a process that may be disturbed at sites of inflammation and in atherosclerotic plaques. In this article, we report our evaluations of the effects of the inflammatory mediator, tumor necrosis factor-alpha (TNF-alpha), on the production of prostacyclin and the barrier function of human endothelial cell monolayers in an in vitro model. TNF-alpha (500 units/ml) had no direct effect on the passage of sucrose, peroxidase, and low density lipoprotein through monolayers of human aortic endothelial cells. On the other hand, during the first hours after addition 500 units/ml TNF-alpha induced a reduction of the permeability of umbilical artery endothelial cell monolayers. Within 10 minutes TNF-alpha induced an increase in prostacyclin production by primary cultures of umbilical artery endothelial cells. However, the reduction in permeability was not caused by a change in prostacyclin production or by a change in cyclic AMP concentration because 1) the effect of TNF-alpha on permeability was not prevented by aspirin, 2) no change in the cellular cyclic AMP concentration could be observed after addition of TNF-alpha, and 3) TNF-alpha was still able to reduce the passage rate in the presence of 25-mu-M forskolin. The reduction in permeability was accompanied by a decrease of F-actin in stress fibers. With prolonged incubation with TNF-alpha, the permeability of umbilical artery endothelial cell monolayers increased, and F-actin was found again in stress fibers. However, these effects of TNF-alpha were only significant at high concentrations of TNF-alpha. Because high TNF-alpha concentrations only persist in vivo for short periods and no increase in the permeability of human aortic endothelial permeability was observed after incubation with TNF-alpha, it is unlikely that TNF-alpha plays a role in the increased permeability that has been found in human arteriosclerotic lesions