Top

Graefe's Archive for Clinical and Experimental Ophthalmology

Published in:

01-02-2009 | Basic Science

The effect of clonidine on VEGF expression in human retinal pigment epithelial cells (ARPE-19)

Authors: Kazuhiko Watanabe, Xue-Yun Zhang, Kiyotaka Kitagawa, Tatsuya Yunoki, Atsushi Hayashi

Published in: Graefe's Archive for Clinical and Experimental Ophthalmology | Issue 2/2009

Abstract

Background

The purpose of this study was to investigate the effect of clonidine, an alpha₂-adrenergic receptor (α₂-ADR) agonist, on vascular endothelial growth factor (VEGF) expression and secretion in the human retinal pigment epithelial cell line (ARPE-19) stimulated with interleukin-1β (IL-1β).

Methods

Alpha₂-ADRs (α₂A, α₂B, and α₂C) mRNA expression in ARPE-19 cells was examined by semiquantitative reverse transcription polymerase chain reaction (RT-PCR). Clonidine and inhibitors against protein kinases that are involved in the regulation of the intracellular signal transduction were added to serum-free medium before stimulation of IL-1β. The α₂-ADR antagonist, Yohimbine, was loaded 30 min before the addition of clonidine. The expression of VEGF mRNA and protein was measured by real-time PCR and enzyme-linked immunosorbent assay.

Results

Alpha₂A-ADR, α₂B-ADR, and α₂C-ADR mRNA was expressed in RPE cells. Clonidine, an inhibitor of p38MAPK and MEK1/2, inhibited the expression of VEGF protein and mRNA in the RPE cells stimulated with IL-1β. The inhibitory effect of clonidine on the secretion of VEGF protein stimulated with IL-1β was blocked by α₂-ADR antagonists.

Conclusions

The effect of clonidine on the expression of VEGF may be via suppression of the p38MAPK and MEK1/2 signal transduction pathways activated with IL-1β.

Henkind P, Gartner S (1983) The relationship between retinal pigment epithelium and the choriocapillaris. Trans Ophthalmol Soc U K 103:444–447PubMed

Yang QR, Smets RME, Neetens A, Berghe DV (1993) Human retinal pigment epithelial cells from different donors continuously produce a vascular endothelial cell-stimulating factor into serum-free medium. J Cell Sci 104:211–218PubMed

Strauss O (2005) The retinal pigment epithelium in visual function. Physiol Rev 85:845–881. doi:10.1152/physrev.00021.2004 PubMedCrossRef

Marano RJ, Rakoczy PE (2005) Treatments for choroidal and retinal neovascularization: a focus on oligonucleotide therapy and delivery for the regulation of gene function. Clin Experiment Ophthalmol 33:81–89. doi:10.1111/j.1442-9071.2005.00952.x PubMedCrossRef

Leung DW, Cachianes G, Kuang WJ, Goeddel DV, Ferrara N (1989) Vascular endothelial growth factor is a secreted angiogenic mitogen. Science 246:1306–1309. doi:10.1126/science.2479986 PubMedCrossRef

Ferrara N (1999) Vascular endothelial growth factor: molecular and biological aspects. Curr Top Microbiol Immunol 237:1–30PubMed

Aiello LP (1997) Vascular endothelial growth factor. 20th-century mechanisms, 21st-century therapies. Invest Ophthalmol Vis Sci 38:1647–1652PubMed

Schlingemann RO, van Hinsbergh VWM (1997) Role of vascular permeability factor/vascular endothelial growth factor in eye disease. Br J Ophthalmol 81:501–512PubMedCrossRef

Mousa SA, Lorelli W, Campochiaro PA (1999) Role of hypoxia and extracellular matrix-integrin binding in the modulation of angiogenic growth factors secretion by retinal pigmented epithelial cells. J Cell Biochem 74:135–143. doi:10.1002/(SICI)1097-4644(19990701)74:1<135::AID-JCB15>3.0.CO;2-# PubMedCrossRef

10.

Lu M, Kuroki M, Amano S, Tolentino M, Keough K, Kim I, Bucala R, Adamis AP (1998) Advanced glycation end products increase retinal vascular endothelial growth factor expression. J Clin Invest 101:1219–1224. doi:10.1172/JCI1277 PubMedCrossRef

11.

Oh H, Takagi H, Takagi C, Suzuma K, Otani A, Ishida K, Matsumura M, Ogura Y, Honda Y (1999) The potential angiogenic role of macrophages in the formation of choroidal neovascular membranes. Invest Ophthalmol Vis Sci 40:1891–1898PubMed

12.

Dinarello CA (1994) The interleukin-1 family: 10 years of discovery. FASEB J 8:1314–1325PubMed

13.

Awad BE, Kreft B, Wolber E-M et al (2000) Hypoxia and IL-1β stimulate vascular endothelial growth factor production in human proximal tubular cells. Kidney Int 58:43–50. doi:10.1046/j.1523-1755.2000.00139.x PubMedCrossRef

14.

Ben-Av P, Crofford LJ, Wilder RL et al (1995) Induction of vascular endothelial growth factor expression in synovial fibroblasts by prostaglandin E and LL-1: a potential mechanism for inflammatory angiogenesis. FEBS Lett 372:83–87. doi:10.1016/0014-5793(95)00956-A PubMedCrossRef

15.

Jung YD, Liu W, Reinmuth N et al (2001) Vascular endothelial growth factor is upregulated by IL-1β in human vascular smooth muscle cells via the p38 mitogen-activated protein kinase pathway. Angiogenesis 4:155–162. doi:10.1023/A:1012291524723 PubMedCrossRef

16.

Kern TS (2007) Contributions of inflammatory processes to the development of the early stages of diabetic retinopathy. Exp Diabetes Res 2007:95103. doi:10.1155/2007/95103 PubMed

17.

Kowluru RA, Odenbach S (2004) Role of IL-1β in the development of retinopathy in rats: effect of antioxidants. Invest Ophthalmol Vis Sci 45:4161–4166. doi:10.1167/iovs.04-0633 PubMedCrossRef

18.

Tatar O, Adam A, Shinoda K, Yoeruek E, Szurman P, Bopp S, Eckardt C, Bartz-Schmidt KU, Grisanti S (2007) Influence of verteporfin photodynamic therapy on inflammation in human choroidal neovascular membranes secondary to age-related macular degeneration. Retina 27:713–723. doi:10.1097/IAE.0b013e318042d3b0 PubMedCrossRef

19.

Patel M, Chan CC (2008) Immunopathological aspects of age-related macular degeneration. Semin Immunopathol 30:97–110. doi:10.1007/s00281-008-0112-9 PubMedCrossRef

20.

Gavras I, Manolis AJ, Gavras H (2001) The α₂-ADRs in hypertension and heart failure: experimental and clinical studies. J Hypertens 19:2115–2124. doi:10.1097/00004872-200112000-00001 PubMedCrossRef

21.

Pollack IP, Brown RH, Crandall AS, Robin AL, Stewart RH, White GL (1988) Effectiveness of apraclonidine in preventing the rise in intraocular pressure after neodymium:YAG posterior capsulotomy. Trans Am Ophthalmol Soc 86:461–472PubMed

22.

Elena PP, Kosina-Boix M, Denis P, Moulin G, Lapalus P (1989) Alpha2-adrenergic receptors in rat and rabbit eye: a tritium-sensitive film autoradiography. Ophthalmic Res 21:309–314PubMedCrossRef

23.

Szelenyi J, Kiss JP, Puskas E, Szelenyi M, Vizi ES (2000) Contribution of differently localized α₂- and β-ADRs in the modulation of TNF-α and IL-10 production in endotoxemic mice. Ann N Y Acad Sci 917:145–153PubMedCrossRef

24.

Liu B, Eisenach JC (2006) Perineural clonidine reduces p38 mitogen-activated protein kinase activation in sensory neurons. Neuroreport 17:1313–1317. doi:10.1097/01.wnr.0000227995.45917.f5 PubMedCrossRef

25.

Tanaka T, Kanai H, Sekiguchi K, Aihara Y, Yokoyama T, Arai M (2000) Induction of VEGF gene transcription by IL-1β is mediated through stress-activated MAP kinases and Sp1 sites in cardiac myocytes. J Mol Cell Cardiol 32:1955–1967. doi:10.1006/jmcc.2000.1228 PubMedCrossRef

26.

Philipp M, Brede ME, Hadamek K, Gessler M, Lohse MJ, Hein L (2002) Placental alpha(2)-adrenoceptors control vascular development at the interface between mother and embryo. Nat Genet 31:311–315. doi:10.1038/ng919 PubMedCrossRef

27.

Jackson JR, Minton JA, Ho ML, Wei N, Winkler JD (1997) Expression of vascular endothelial growth factor in synovial fibroblasts is induced by hypoxia and IL-1β. J Rheumatol 24:1253–1259PubMed

28.

Jung YJ, Isaacs JS, Sunmin L, Trepel J, Neckers L (2003) IL-1β-mediated up-regulation of HIF-1α via an NFκB/COX-2 pathway identifies HIF-1 as a critical link between inflammation and oncogenesis. FASEB J 17:2115–2117PubMed

29.

Roberge FG, Caspi RR, Nussenblatt RB (1988) Glial retinal Mueller cells produce II-1 activity and have a dual effect on autoimmune T helper lymphocytes. J Immunol 140:2193–2196PubMed

30.

Helbig H, Kittredge KL, Gurley RC, Thurau SR, Palestine AG, Nussenblatt RB (1990) Endotoxin-induced production of inflammatory mediators by cultured ciliary epithelial cells. Curr Eye Res 9:501–505. doi:10.3109/02713689008999616 PubMedCrossRef

31.

Vinores SA, Xiao WH, Zimmerman R, Whitcup SM, Wawrousek EF (2003) Upregulation of vascular endothelial growth factor (VEGF) in the retinas of transgenic mice overexpressing IL-1β in the lens and mice undergoing retinal degeneration. Histol Histopathol 18:797–810PubMed

32.

Demircan N, Safran BG, Soylu M, Ozcan AA, Sizmaz S (2006) Determination of vitreous IL-1 and TNF-α levels in proliferative diabetic retinopathy. Eye 20:1366–1369. doi:10.1038/sj.eye.6702138 PubMedCrossRef

33.

Yoshino Y, Aoyagi M, Tamaki M, Duan L, Morimoto T, Ohno K (2006) Activation of p38 MAPK and/or JNK contributes to increased levels of VEGF secretion in human malignant glioma cells. Int J Oncol 29:981–987PubMed

34.

Kable JW, Murrin LC, Bylund DB (2000) In vivo gene modification elucidates subtype-specific functions o f α₂-ADRs. J Pharmacol Exp Ther 293:1–7PubMed

35.

Platts KE, Benson MT, Rennie IG, Sharrard RM, Rees RC (1995) Cytokine modulation of adhesion molecule expression on human retinal pigment epithelial cells. Invest Ophthalmol Vis Sci 36:2262–2269PubMed

36.

Campochiaro PA, Sugg R, Grotendorst G, Hjelmeland LM (1989) Retinal pigment epithelial cells produce PDGF-like proteins and secrete them into their media. Eye Exp Res 49:217–227. doi:10.1016/0014-4835(89)90092-4 CrossRef

37.

Elner VM, Strieter RM, Elner SG (1990) Neutrophil chemotactic factor (IL-8) gene expression by cytokine-treated retinal pigment epithelial cells. Am J Pathol 136:745–750PubMed

38.

Holtkamp GM, De Vos AF, Peek R, Kijlsta A (1999) Analysis of the secretion pattern of monocyte chemotactic protein-1 (MCP-1) and transforming growth factor-beta 2 (TGF-β2) by human retinal pigment epithelial cells. Clin Exp Immunol 118:35–40. doi:10.1046/j.1365-2249.1999.01016.x PubMedCrossRef

39.

Kanga BY, Leeb SW, Kim TS (2003) Stimulation of interleukin-12 production in mouse macrophages via activation of p38 mitogen-activated protein kinase by α₂-ADRs agonists. Eur J Pharmacol 467:223–231. doi:10.1016/S0014-2999(03)01628-5 CrossRef

40.

Romero-Sandoval A, Eisenach JC (2006) Perineural clonidine reduces mechanical hypersensitivity and cytokine production in established nerve injury. Anesthesiology 104:351–355. doi:10.1097/00000542-200602000-00022 PubMedCrossRef

41.

Xu B, Makris A, Thornton C, Ogle R, Horvath J, Hennessy A (2006) Antihypertensive drugs clonidine, diazoxide, hydralazine and furosemide regulate the production of cytokines by placentas and peripheral blood mononuclear cells in normal pregnancy. Pregnancy 24:915–922

Title: The effect of clonidine on VEGF expression in human retinal pigment epithelial cells (ARPE-19)
Authors: Kazuhiko Watanabe
Xue-Yun Zhang
Kiyotaka Kitagawa
Tatsuya Yunoki
Atsushi Hayashi
Publication date: 01-02-2009
Publisher: Springer-Verlag
Published in: Graefe's Archive for Clinical and Experimental Ophthalmology / Issue 2/2009
Print ISSN: 0721-832X
Electronic ISSN: 1435-702X
DOI: https://doi.org/10.1007/s00417-008-0990-5

Keynote webinar | Spotlight on medication adherence

Springer Medicine

The effect of clonidine on VEGF expression in human retinal pigment epithelial cells (ARPE-19)

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 2/2009

Minimally invasive strabismus surgery (MISS) for inferior obliquus recession

Intravitreal administration of the anti-TNF monoclonal antibody Infliximab in the rabbit

Physiology of vitreous surgery

Quantitative analysis of central visual field defects in macular edema using three-dimensional computer-automated threshold Amsler grid testing

Diabetic retinopathy and health-related quality of life

Influence of cosmetically tinted soft contact lenses on higher-order wavefront aberrations and visual performance