Endogenous prostaglandin D2 synthesis inhibits e-selectin generation in human umbilical vein endothelial cells

DOI: 10.4236/health.2011.35053   PDF   HTML     4,922 Downloads   8,268 Views  


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.

Share and Cite:

Negoro, H. , Kobayashi, H. and Uehara, Y. (2011) Endogenous prostaglandin D2 synthesis inhibits e-selectin generation in human umbilical vein endothelial cells. Health, 3, 304-311. doi: 10.4236/health.2011.35053.

Conflicts of Interest

The authors declare no conflicts of interest.


[1] 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.
[2] 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
[3] 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
[4] 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
[5] 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.
[6] 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.
[7] 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
[8] 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.
[9] 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
[10] 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.
[11] Chen, C. and Okayama, H. (1987) High-efficiency transformation of mammalian cells by plasmid DNA. Molecular and Cellular Biology, 7, 2745-2752.
[12] 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
[13] 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
[14] 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
[15] 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.
[16] 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
[17] 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.
[18] 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.
[19] 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
[20] 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.
[21] 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
[22] 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
[23] 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
[24] 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
[25] 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

comments powered by Disqus

Copyright © 2020 by authors and Scientific Research Publishing Inc.

Creative Commons License

This work and the related PDF file are licensed under a Creative Commons Attribution 4.0 International License.