抄録
For the simple examination of the change in vascular permeability response of endothelial cells to various stimuli, we attempted to establish an in vitro experimental model using Boyden's chamber. This chamber has both upper and lower compartments with a partition filter between them. We succeeded in obtaining a monolayer of vascular endothelial cells on the surface of the filter by using a gelatin coated nucleopore filter. Both upper and lower compartments were filled with culture medium. When bovine serum albumin was added only to the upper compartment, the albumin was time-dependently leaked from the upper compartment to the lower compartment though the pores of the filter. The leakage was inhibited greatly when the surface of the filter was covered with entothelial cells.
By monitoring the amount of albumin leaked into the lower compartment after the addition of various stimuli, such as histamine, serotonin, bradykinin, PAF (Platelet activating factor) etc., to the upper compartment, we could judge which stimulus could better influence vascular permeability. PAF was found to be the most active stimulus in increasing vascular permeability.
During the course of the investigation, we found that such substances as having cytotoxic activity, i.e., lipid peroxides, antiserum in the presence of the complement etc., could also increase vascular permeability. Therefore, by measuring cell viability at the same time, this model can be used as an in vitro assay system for measuring vascular endothelial cell injury. This model may also be a useful in vitro assay system for the evaluation of cytoprotective drugs. By using this system we found that 15-HPETE (15-hydroxyeicosatetraenoic acid), one of the lipoxygenase metabolites of arachidonic acid, has a strong cytolitic activity on vascular endothelial cells and that MCI-186 (3-methyl-1-phenyl-2-pyrazolin-5-one) could prevent the cells from such injury.
