We examined the role of prostaglandin D2 (PGD2) in the formation of E-selectin following inter-leukin-1 (IL-1) stimulation in human umbilical vein endothelial cells (HUVEC) transfected with lipocaline-type PGD2 synthase (L-PGDS) genes. HUVEC were isolated from human umbilical vein and incubated with 20 U/mL IL-1 and various concentrations of authentic PGD2. The isolated HUVEC were also transfected with L-PGDS genes by electroporation. The L-PGDS-transfected HUVEC were used to investigate the role of endogenous PGD2 in IL-1-stimulated E-selectin biosynthesis. We also used an anti-PGD2 antibody to examine whether an intracrine mechanism was involved in E-selectin production. PGD2 and E-selectin levels were determined by radio-immunoassay and enzyme- immunoassay, respectively. E-selectin mRNA was assessed by real-time RT-PCR. IL-1-stimulated E-selectin production by HUVEC was dose-dependently inhibited by authentic PGD2 at concentrations greater than 10–6 mol/L. L-PGDS gene-transfected HUVEC produced more PGD2 than HUVEC transfected with the reporter gene alone. IL-1 induced increases in E-selectin production in HUVEC transfected with the reporter genes alone. However, this effect was significantly attenuated in the case of IL-1 stimulation of HUVEC trans-fected with L-PGDS genes, and accompanied by an apparent suppression of E-selectin mRNA expression. Neutralization of extracellular PGD2 by anti-PGD2- specific antibody influenced neither E-selectin mRNA expression nor E-selectin biosynthesis. HUVEC transfected with L-PGDS genes showed increased PGD2 synthesis. This increase was associated with attenuation of both E-selectin generation and E-selectin mRNA expression. The results suggest that endogenous PGD2 decreases E-selectin synthesis and E-selectin mRNA expression, probably through an intracrine mechanism.
Joris, I., Zand, T., Nuunari, J.J., Krolikowski, F.J. and Majno, G. (1983) Studies on the pathogenesis of atherosclerosis, I: Adhesion and emigration of mononuclear cells in the aorta of hypercholesterolemic rats. American Journal of Pathology, 113, 341-358.
Rao, R.M., Yang, L., Garcia-Cardena, G. and Luscinskas, F.W. (2007) Endothelial-development mechanisms of leukocytes recruitment to the vascular wall. Circulation Research, 101, 234-247. doi:10.1161/CIRCRESAHA.107.151860b
Cybulsky, M.I. and Gimbrone, M.A.Jr. (1991) Endothelial expression of a mononuclear leukocyte adhesion molecule, in rabbit aortic endothelium. Science, 251, 788-791. doi:10.1126/science.1990440
Tousoulis, D., Antoniades, C., Bosinakou, E., Kotu, M., Tsoufis C., Marinou, K., et al. (2007) Differences in inflammatory and thrombotic markers between unstable angina and acute myocardial infarction. International Journal of Cardiology, 115, 203-207. doi:10.1016/j.ijcard.2006.03.011
Wal, van der A.C., Das, P.K., Tigges, A.J. and Becker, A.E. (1992) Adhesion molecules on the endothelium and mononuclear cells in human atherosclerotic lesions. American Journal of Pathology, 141, 1427-1433.
Nagoshi, H., Uehara, Y., Kanai, F., Maeda, S., Ogura, T., Goto, A. and Omata, M. (1998) Prostaglandin D2 inhibits inducible nitric oxide synthase expression in rat smooth muscle cells. Circulation Research, 82, 204-209.
Negoro, H., Shin, W.S., Taguchi, R.H., Eguchi, N., Urade, Y., Toyo-oka, T., Fujita, T., Omata, M. and Uehara, Y., (2002) Endogenous prostaglandin D2 synthesis reduces an increase in plasminogen activator inhibitor-1 following interleukin stimulation in bovine endothelial cells. Journal of Hypertension, 20, 1347-1354. doi:10.1097/00004872-200207000-00021
Watanabe, T., Narumiya, S., Shimizu, T. and Hayaishi, O. (1982) Characterization of the biosynthetic pathway of prostaglandin D2 in human platelet-rich plasma. Journal of Biological Chemistry, 257, 14847-14853.
Eguchi, Y., Eguchi, N., Oda, H., Seiki, K., Kijima and Y., Matsu-ura, Y. (1997) Expression of lipocalin-type pros- taglandin D synthase (beta-trace) in human heart and its accumulation in the coronary circulation of angina pa- tients. Proceedings of National Academy of Sciences of the United States of America, 94, 14689-14694. doi:10.1073/pnas.94.26.14689
Urade, Y., Nagata, A., Suzuki, Y., Fujii, Y. and Hayaishi, O. (1989) Primary structure of rat brain prostaglandin D synthase deducted from cDNA sequence. Journal of Bio- logical Chemistry, 264, 1041-1045.
Chen, C. and Okayama, H. (1987) High-efficiency transformation of mammalian cells by plasmid DNA. Molecular and Cellular Biology, 7, 2745-2752.
Zimmerman, G.A., Whatley, R.E., McIntyre, T.M., Benson, D.E. and Prescott, S.M. (1990) Endothelial cells for studies of platelet-activating factor and arachidonate metabolites. Methods in Enzymology, 187, 520-535. doi:10.1016/0076-6879(90)87059-C
Ohtani, K., Nakamura, M., Saito, S., Nagata, K., Sugamura, K. and Hinuma, Y. (1989) Electroporation: Application to human lymphoid cell lines for stable introduction of a transactivator gene of human T-cell leukemia virus type 1. Nucleic Acid Research, 17, 1589-1604. doi:10.1093/nar/17.4.1589
Bonnerot, C., Rocancourt, D., Briant, P., Grimber, G. and Nicolas, J.F. (1987) A b-galactosidase hybrid protein tar- geted to nuclei as a marker for developmental studies. Proceedings of National Academy of Sciences of the United States of America, 84, 6795-6799. doi:10.1073/pnas.84.19.6795
Tobian, L., Uehara, Y. and Iwai, J. (1985) Prostaglandin alterations in barely hypertensive Dahl S rats. Trans Association of the American Physiology, 98, 378-383.
Uehara, Y., Tobian, L., Iwai, J., Ishii, M. and Sugimoto, T. (1987) Alterations of vascular prostacyclin and thromboxane A2 in Dahl genetical strain susceptible to salt-induced hypertension. Prostaglandins, 33, 727-738. doi:10.1016/0090-6980(87)90038-4
Koide, M., Kawahara, Y., Nakayama, I., Tsuda, T. and Yokoyama, M. (1993) Cyclic AMP-elevation agents induce an inducible type of nitric oxide synthase in cultured vascular smooth muscle cells. Journal of Biological Chemistry, 268, 24959-24966.
Tamasawa, N., Murakami, H., Matsui, J., Yamato, K., JingZhi, G., Imaizumi, T., Fujimoto, K., Yoshida, H., Satoh, K. and Suda, T. (2001) An oxidised derivative of cholesterol increase the release of soluble vascular cell adhesion molecule-1 from human umbilical vein endothelial cells in culture. Biochimica et Biophysica Acta-General Subjects, 1531, 178-187.
Negoro, H., Shin, W.S., Hakamada-Taguchi, R., Eguchi, N., Urade, Y., Goto, A., Toyo-Oka, T., Fujita, T., Omata, M. and Uehara, Y. (2005) Endogenous prostaglandin D(2) synthesis decreases vascular cell adhesion molecule-1 expression in human umbilical vein endothelial cells. Life Sciences, 78, 22-29. doi:10.1016/j.lfs.2005.02.037
Zidovetzki, R., Wang, J.L., Kim, J.A., Chen, P., Fisher, M. and Hofman, F.M. (1999) Endothelin-1 enhances plasminogen activator-1 production by human brain endothelial cells via protein kinase C-dependent pathway. Arteriosclerosis Thrombosis and Vascular Biology, 19, 1768- 1775.
Forman, B.M., Tontonoz, P., Chen, J., Brun, R.P., Spiegelman, B.M. and Evans, R.M. (1995) 15-Deoxy- D12,14-prostaglandin J2 is a ligand for the adipocyte determination factor PPAR. Cell, 83, 803-812. doi:10.1016/0092-8674(95)90193-0
Kliewer, S.A., Lenhard, J.M., Willson, T.M., Patel, I., Morris, D.C. and Lehmann, J.M. (1995) A prostaglandin J2 metabolite binds peroxysome proliferator-activator g and promotes adipocyte differentiation. Cell, 83, 813- 819. doi:10.1016/0092-8674(95)90194-9
Kikwa, Y., Narumiya, S., Fukushima, M., Wakatsuka, H. and Hayaish, O. (1984) 9-deoxy-d9,d12-13,14-dihydroprostaglandin D2, a metabolite of prostaglandin D2 formed in human plasma. Proceedings of National Academy of Sciences of the United States of America, 81, 1317-1321. doi:10.1073/pnas.81.5.1317
Ricote, M., Li, A.C., Willson, T.M., Kelly, C.J. and Glass, C.K. (1998) The peroxysome proliferator-activated receptor-g is a negative regulator of macrophage activation. Nature, 391, 79-82. doi:10.1038/34178
Yu, G., Rux, A.H., Ma, P., Bdeir, K. and Sachais, B.S. (2005) Endothelial expression of E-selectin is induced by the platelet-specific chemokine platelet factor 4 through LRP in an NF-kappaB-dependent manner. Blood, 105, 3545-3551. doi:10.1182/blood-2004-07-2617