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Negoro, H. et al.

Negoro, Soo Shin, Hakamada-Taguchi, Eguchi, Urade, Goto, Toyo-Oka, Fujita, Omata, Uehara,

Endogenous prostaglandin D2 synthesis reduces an increase in plasminogen activator inhibitor-1 following interleukin stimulation in bovine endothelial cells.

OBJECTIVE : We examined the role of prostaglandin D2 (PGD2) in the formation of plasminogen activator inhibitor (PAI)-1 following interleukin-1beta (IL-1) stimulation in bovine endothelial cells (EC) transfected with lipocaline-type PGD2 synthase (L-PGDS) genes. DESIGN AND METHODS : EC were isolated from bovine thoracic aorta and incubated with 20 U/ml IL-1 and various concentrations of authentic PGD2. The isolated EC were also transfected with L-PGDS genes by electroporation. The L-PGDS-transfected EC were used to investigate the role of endogenous PGD2 in IL-1 stimulated PAI-1 biosynthesis. We also used an anti-PGD2 antibody to examine whether an intracrine mechanism was involved in PAI-1 production. PGD2 and PAI-1 levels were determined by radio- and enzyme-immunoassay, respectively. PAI-1 mRNA was assessed by reverse transcription-polymerase chain reaction. RESULT : IL-1 stimulated PAI-1 production by EC was dose-dependently inhibited by authentic PGD2 at concentrations greater than 10-6 mol/l. L-PGDS gene-transfected EC produced more PGD2 than EC transfected with the reporter gene alone. IL-1 induced increases in PAI-1 production in EC transfected with reporter genes alone. However, this effect was significantly attenuated in the case of IL-1 stimulation of EC transfected with L-PGDS genes, and accompanied by an apparent suppression of PAI-1 mRNA expression. The effects of PGD2 on PAI-I formation were reversed to the basal levels by the inhibition of synthesis of endogenous PGD2. Neutralization of extracellular PGD2 by anti-PGD2 antibody influenced neither PAI-1 mRNA expression nor PAI-1 biosynthesis. CONCLUSION : EC transfected with L-PGDS genes increased PGD2 synthesis. This was associated with attenuation of both PAI-1 formation and PAI-1 mRNA expression. It is suggested that endogenous PGD2 decreases PAI-1 synthesis and PAI-1 mRNA expression, probably through an intracrine mechanism.[1]

References

Endogenous prostaglandin D2 synthesis reduces an increase in plasminogen activator inhibitor-1 following interleukin stimulation in bovine endothelial cells. Negoro, H., Soo Shin, W., Hakamada-Taguchi, R., Eguchi, N., Urade, Y., Goto, A., Toyo-Oka, T., Fujita, T., Omata, M., Uehara, Y. J. Hypertens. (2002) [Pubmed]

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