Genetic deletion of COX-2 diminishes VEGF production in mouse retinal Müller cells.
Non-steroidal anti-inflammatory drugs (NSAIDs), which inhibit COX activity, reduce the production of retinal VEGF and neovascularization in relevant models of ocular disease. We hypothesized that COX-2 mediates VEGF production in retinal Müller cells, one of its primary sources in retinal neovascular disease. The purpose of this study was to determine the role of COX-2 and its products in VEGF expression and secretion. These studies have more clearly defined the role of COX-2 and COX-2-derived prostanoids in retinal angiogenesis. Müller cells derived from wild-type and COX-2 null mice were exposed to hypoxia for 0-24 h. COX-2 protein and activity were assessed by western blot analysis and GC-MS, respectively. VEGF production was assessed by ELISA. Wild-type mouse Müller cells were treated with vehicle (0.1% DMSO), 10 microM PGE(2), or PGE(2) + 5 microM H-89 (a PKA inhibitor), for 12 h. VEGF production was assessed by ELISA. Hypoxia significantly increased COX-2 protein (p < 0.05) and activity (p < 0.05), and VEGF production (p < 0.0003). COX-2 null Müller cells produced significantly less VEGF in response to hypoxia (p < 0.05). Of the prostanoids, PGE(2) was significantly increased by hypoxia (p < 0.02). Exogenous PGE(2) significantly increased VEGF production by Müller cells (p < 0.0039), and this effect was inhibited by H-89 (p < 0.055). These data demonstrate that hypoxia induces COX-2, prostanoid production, and VEGF synthesis in Müller cells, and that VEGF production is at least partially COX-2-dependent. Our study suggests that PGE(2), signaling through the EP(2) and/or EP(4) receptor and PKA, mediates the VEGF response of Müller cells.
Yanni
Susan E
SE
Department of Cell and Developmental Biology, Vanderbilt University School of Medicine, Nashville, TN 37232-8808, USA. Susan.e.yanni@gmail.com
McCollum
Gary W
GW
Penn
John S
JS
D000818:Animals; D000831:Animals, Newborn; D015153:Blotting, Western; D002478:Cells, Cultured; D017868:Cyclic AMP-Dependent Protein Kinases / Q000037:antagonists & inhibitors; D051546:Cyclooxygenase 2 / Q000502:physiology*; D015232:Dinoprostone / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D008401:Gas Chromatography-Mass Spectrometry; D017353:Gene Deletion; D000860:Hypoxia / Q000378:metabolism; D007546:Isoquinolines / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D017628:Microglia / Q000378:metabolism*; D011453:Prostaglandins / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D055351:Retinal Neurons / Q000378:metabolism*; D013449:Sulfonamides / Q000494:pharmacology; D042461:Vascular Endothelial Growth Factor A / Q000096:biosynthesis*
D007546:Isoquinolines; D011453:Prostaglandins; D047428:Protein Kinase Inhibitors; D013449:Sulfonamides; D042461:Vascular Endothelial Growth Factor A; C467480:vascular endothelial growth factor A, mouse; C077623:Ptgs2 protein, mouse; D051546:Cyclooxygenase 2; D017868:Cyclic AMP-Dependent Protein Kinases; D015232:Dinoprostone; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide
Invest Ophthalmol Vis Sci. 2006 Nov;47(11):5047-56
17065526
Exp Eye Res. 1990 Aug;51(2):119-29
2387332
Cancer Metastasis Rev. 2007 Dec;26(3-4):525-34
17763971
Invest Ophthalmol Vis Sci. 2003 Jan;44(1):409-15
12506103
Mol Vis. 2006 Apr 20;12:405-14
16688111
Life Sci. 2003 Dec 5;74(2-3):143-53
14607241
Cancer Res. 2006 Jan 1;66(1):143-50
16397226
Invest Ophthalmol Vis Sci. 2001 Oct;42(11):2616-24
11581208
J Biol Chem. 2003 Oct 3;278(40):38607-16
12874281
J Biol Chem. 2000 Jan 28;275(4):2837-44
10644750
Growth Factors. 1997;14(4):229-41
9386988
Curr Eye Res. 1999 Oct;19(4):300-4
10520225
J Biol Chem. 1997 Jan 3;272(1):601-8
8995303
Jpn J Pharmacol. 2001 Nov;87(3):181-8
11885966
Proc Soc Exp Biol Med. 1983 Feb;172(2):214-8
6572402
Eur J Pharmacol. 2003 Jan 5;458(3):283-9
12504784
Brain Res Bull. 2010 Feb 15;81(2-3):310-9
19897019
J AAPOS. 1999 Feb;3(1):26-32
10071898
Biochem Biophys Res Commun. 2001 Sep 7;286(5):923-8
11527387
Neoplasia. 2001 Jan-Feb;3(1):53-61
11326316
J Biol Chem. 2005 Aug 26;280(34):29993-30000
15970595
Int J Biochem Cell Biol. 1999 May;31(5):551-7
10399316
Arch Ophthalmol. 1995 Dec;113(12):1538-44
7487623
Biochem Biophys Res Commun. 2004 Dec 10;325(2):461-6
15530415
Growth Factors. 1998;16(1):1-9
9777366
Pharmacol Ther. 2004 Aug;103(2):147-66
15369681
Hum Cell. 2002 Mar;15(1):13-24
12126060
J Clin Invest. 2000 Jun;105(11):1589-94
10841517
Cancer Res. 1998 Feb 1;58(3):409-12
9458081
J Neurosci Res. 1999 Oct 15;58(2):254-61
10502281
Ophthalmology. 1996 Nov;103(11):1721-6
8942862
Mol Carcinog. 2007 Nov;46(11):912-23
17427962
Invest Ophthalmol Vis Sci. 1994 Nov;35(12):3974-9
7525506
J Pathol. 2009 Dec;219(4):446-54
19768732
Exp Eye Res. 2004 Aug;79(2):275-85
15325574
Mol Pharmacol. 2005 Aug;68(2):317-29
15883203
Eur J Pharmacol. 2004 Sep 13;498(1-3):275-8
15364005
FASEB J. 1998 Sep;12(12):1063-73
9737710
Inflammopharmacology. 2007 Oct;15(5):214-7
17943254
Cancer Res. 2006 Jul 1;66(13):6683-91
16818642
Am J Respir Cell Mol Biol. 1997 Dec;17(6):748-56
9409562
Histol Histopathol. 1994 Oct;9(4):807-43
7534514
Proc Natl Acad Sci U S A. 1995 Jan 31;92(3):905-9
7846076
Invest Ophthalmol Vis Sci. 2006 Mar;47(3):1149-60
16505053
Neurosci Lett. 2007 Jun 21;421(1):22-6
17548159
Invest Ophthalmol Vis Sci. 2003 Aug;44(8):3606-13
12882814
Retina. 2005 Dec;25(8):1054-64
16340537
Invest Ophthalmol Vis Sci. 2003 Mar;44(3):974-9
12601017
Invest Ophthalmol Vis Sci. 2009 Nov;50(11):5479-86
19494202
J Comp Neurol. 2002 Oct 28;452(4):392-9
12355421
Circulation. 2003 Jul 15;108(2):198-204
12821538
Invest Ophthalmol Vis Sci. 1998 Mar;39(3):581-91
9501870
Arthritis Rheum. 2004 Aug;50(8):2441-9
15334456
Biochem Biophys Res Commun. 2003 Jul 11;306(4):887-97
12821125
Invest Ophthalmol Vis Sci. 1997 Aug;38(9):1647-52
9286253
J Natl Cancer Inst. 1982 Aug;69(2):475-82
6180207
Proc Natl Acad Sci U S A. 2004 Jan 13;101(2):415-6
14707264
Oncogene. 1999 Dec 20;18(55):7908-16
10630643
Surv Ophthalmol. 1998 Nov-Dec;43(3):245-69
9862312
Am J Ophthalmol. 1997 Oct;124(4):521-9
9323943
Exp Eye Res. 2001 Jun;72(6):611-21
11384149
J Immunol. 2004 Jan 15;172(2):1227-36
14707101