Nicotine increases the VEGF/PEDF ratio in retinal pigment Epithelium: A possible mechanism for CNV in passive smokers with AMD

Marianne Pons, Maria E. Marin-Castaño

Research output: Contribution to journalArticle

48 Citations (Scopus)

Abstract

Purpose. Cigarette smoking is the strongest environmental risk factor for wet age-related macular degeneration (AMD). Inappropriate expression of proangiogenic vascular endothelial growth factor (VEGF) and antiangiogenic pigment epithelium derived factor (PEDF) may cause choroidal neovascularization (CNV), a key event in wet AMD, resulting in vision loss. Nicotine (NT), a potent angiogenic agent abundant in second-hand smoke, may play a major role in the pathogenesis of wet AMD. The purpose of this study was to evaluate the expression of nicotinic acetylcholine receptors (nAchR) in retinal pigment epithelium (RPE) and determine the effects of NT on RPE-derived VEGF and PEDF expression in the context of passive smoking. Methods. Human RPE cells were treated with NT (10 -8 M), with or without the nAchR-nonspecific antagonist hexamethonium (HXM) (10 -5 M) for 72 hours. RPE sheets were microdissected from rats exposed to NT in drinking water (100 μg/mL), with or without HXM (40 mg/kg/d, intraperitoneally), for 72 hours. Cell death was determined by cell count and proliferation by Western blot for proliferating cell nuclear antigen (PCNA). nAchR expression was examined by real-time PCR and Western blot. ERK activation was evaluated by Western blot analysis. VEGF and PEDF expression was assessed by ELISA, Western blot, and real-time PCR. Results. Cultured RPE cells constitutively expressed the nAchR α3, α10, and β1 subunits, with β1 being the most prevalent. The nAchR α4, α5, α7, and β2 subunits were detected in RPE sheets from rats, among which α4 is the predominant subtype. NT, which did not result in either cell death or proliferation, induced β1 nAchR, upregulated VEGF, and downregulated PEDF expression through nAChR in ARPE-19 cells. Transcriptional activation of the nAchR α4 subunit and nAChR-mediated upregulation of VEGF and PEDF were observed in RPE from rats exposed to NT. Conclusions. NT increased the VEGF-to-PEDF ratio in the RPE through nAchR in vitro and in vivo. This alteration in the ratio may play a key role in the progression to wet AMD in passive smokers.

Original languageEnglish
Pages (from-to)3842-3853
Number of pages12
JournalInvestigative Ophthalmology and Visual Science
Volume52
Issue number6
DOIs
StatePublished - May 1 2011
Externally publishedYes

Fingerprint

Choroidal Neovascularization
Retinal Pigment Epithelium
Macular Degeneration
Nicotinic Receptors
Nicotine
Vascular Endothelial Growth Factor A
Western Blotting
Hexamethonium
Tobacco Smoke Pollution
Real-Time Polymerase Chain Reaction
Cell Death
Cell Proliferation
pigment epithelium-derived factor
Proliferating Cell Nuclear Antigen
Drinking Water
Transcriptional Activation
Up-Regulation
Down-Regulation
Cell Count
Smoking

ASJC Scopus subject areas

  • Ophthalmology
  • Sensory Systems
  • Cellular and Molecular Neuroscience
  • Medicine(all)

Cite this

Nicotine increases the VEGF/PEDF ratio in retinal pigment Epithelium : A possible mechanism for CNV in passive smokers with AMD. / Pons, Marianne; Marin-Castaño, Maria E.

In: Investigative Ophthalmology and Visual Science, Vol. 52, No. 6, 01.05.2011, p. 3842-3853.

Research output: Contribution to journalArticle

@article{ded0f4c4c3a047bcbbacf6988510a3bd,
title = "Nicotine increases the VEGF/PEDF ratio in retinal pigment Epithelium: A possible mechanism for CNV in passive smokers with AMD",
abstract = "Purpose. Cigarette smoking is the strongest environmental risk factor for wet age-related macular degeneration (AMD). Inappropriate expression of proangiogenic vascular endothelial growth factor (VEGF) and antiangiogenic pigment epithelium derived factor (PEDF) may cause choroidal neovascularization (CNV), a key event in wet AMD, resulting in vision loss. Nicotine (NT), a potent angiogenic agent abundant in second-hand smoke, may play a major role in the pathogenesis of wet AMD. The purpose of this study was to evaluate the expression of nicotinic acetylcholine receptors (nAchR) in retinal pigment epithelium (RPE) and determine the effects of NT on RPE-derived VEGF and PEDF expression in the context of passive smoking. Methods. Human RPE cells were treated with NT (10 -8 M), with or without the nAchR-nonspecific antagonist hexamethonium (HXM) (10 -5 M) for 72 hours. RPE sheets were microdissected from rats exposed to NT in drinking water (100 μg/mL), with or without HXM (40 mg/kg/d, intraperitoneally), for 72 hours. Cell death was determined by cell count and proliferation by Western blot for proliferating cell nuclear antigen (PCNA). nAchR expression was examined by real-time PCR and Western blot. ERK activation was evaluated by Western blot analysis. VEGF and PEDF expression was assessed by ELISA, Western blot, and real-time PCR. Results. Cultured RPE cells constitutively expressed the nAchR α3, α10, and β1 subunits, with β1 being the most prevalent. The nAchR α4, α5, α7, and β2 subunits were detected in RPE sheets from rats, among which α4 is the predominant subtype. NT, which did not result in either cell death or proliferation, induced β1 nAchR, upregulated VEGF, and downregulated PEDF expression through nAChR in ARPE-19 cells. Transcriptional activation of the nAchR α4 subunit and nAChR-mediated upregulation of VEGF and PEDF were observed in RPE from rats exposed to NT. Conclusions. NT increased the VEGF-to-PEDF ratio in the RPE through nAchR in vitro and in vivo. This alteration in the ratio may play a key role in the progression to wet AMD in passive smokers.",
author = "Marianne Pons and Marin-Casta{\~n}o, {Maria E.}",
year = "2011",
month = "5",
day = "1",
doi = "10.1167/iovs.10-6254",
language = "English",
volume = "52",
pages = "3842--3853",
journal = "Investigative Ophthalmology and Visual Science",
issn = "0146-0404",
publisher = "Association for Research in Vision and Ophthalmology Inc.",
number = "6",

}

TY - JOUR

T1 - Nicotine increases the VEGF/PEDF ratio in retinal pigment Epithelium

T2 - A possible mechanism for CNV in passive smokers with AMD

AU - Pons, Marianne

AU - Marin-Castaño, Maria E.

PY - 2011/5/1

Y1 - 2011/5/1

N2 - Purpose. Cigarette smoking is the strongest environmental risk factor for wet age-related macular degeneration (AMD). Inappropriate expression of proangiogenic vascular endothelial growth factor (VEGF) and antiangiogenic pigment epithelium derived factor (PEDF) may cause choroidal neovascularization (CNV), a key event in wet AMD, resulting in vision loss. Nicotine (NT), a potent angiogenic agent abundant in second-hand smoke, may play a major role in the pathogenesis of wet AMD. The purpose of this study was to evaluate the expression of nicotinic acetylcholine receptors (nAchR) in retinal pigment epithelium (RPE) and determine the effects of NT on RPE-derived VEGF and PEDF expression in the context of passive smoking. Methods. Human RPE cells were treated with NT (10 -8 M), with or without the nAchR-nonspecific antagonist hexamethonium (HXM) (10 -5 M) for 72 hours. RPE sheets were microdissected from rats exposed to NT in drinking water (100 μg/mL), with or without HXM (40 mg/kg/d, intraperitoneally), for 72 hours. Cell death was determined by cell count and proliferation by Western blot for proliferating cell nuclear antigen (PCNA). nAchR expression was examined by real-time PCR and Western blot. ERK activation was evaluated by Western blot analysis. VEGF and PEDF expression was assessed by ELISA, Western blot, and real-time PCR. Results. Cultured RPE cells constitutively expressed the nAchR α3, α10, and β1 subunits, with β1 being the most prevalent. The nAchR α4, α5, α7, and β2 subunits were detected in RPE sheets from rats, among which α4 is the predominant subtype. NT, which did not result in either cell death or proliferation, induced β1 nAchR, upregulated VEGF, and downregulated PEDF expression through nAChR in ARPE-19 cells. Transcriptional activation of the nAchR α4 subunit and nAChR-mediated upregulation of VEGF and PEDF were observed in RPE from rats exposed to NT. Conclusions. NT increased the VEGF-to-PEDF ratio in the RPE through nAchR in vitro and in vivo. This alteration in the ratio may play a key role in the progression to wet AMD in passive smokers.

AB - Purpose. Cigarette smoking is the strongest environmental risk factor for wet age-related macular degeneration (AMD). Inappropriate expression of proangiogenic vascular endothelial growth factor (VEGF) and antiangiogenic pigment epithelium derived factor (PEDF) may cause choroidal neovascularization (CNV), a key event in wet AMD, resulting in vision loss. Nicotine (NT), a potent angiogenic agent abundant in second-hand smoke, may play a major role in the pathogenesis of wet AMD. The purpose of this study was to evaluate the expression of nicotinic acetylcholine receptors (nAchR) in retinal pigment epithelium (RPE) and determine the effects of NT on RPE-derived VEGF and PEDF expression in the context of passive smoking. Methods. Human RPE cells were treated with NT (10 -8 M), with or without the nAchR-nonspecific antagonist hexamethonium (HXM) (10 -5 M) for 72 hours. RPE sheets were microdissected from rats exposed to NT in drinking water (100 μg/mL), with or without HXM (40 mg/kg/d, intraperitoneally), for 72 hours. Cell death was determined by cell count and proliferation by Western blot for proliferating cell nuclear antigen (PCNA). nAchR expression was examined by real-time PCR and Western blot. ERK activation was evaluated by Western blot analysis. VEGF and PEDF expression was assessed by ELISA, Western blot, and real-time PCR. Results. Cultured RPE cells constitutively expressed the nAchR α3, α10, and β1 subunits, with β1 being the most prevalent. The nAchR α4, α5, α7, and β2 subunits were detected in RPE sheets from rats, among which α4 is the predominant subtype. NT, which did not result in either cell death or proliferation, induced β1 nAchR, upregulated VEGF, and downregulated PEDF expression through nAChR in ARPE-19 cells. Transcriptional activation of the nAchR α4 subunit and nAChR-mediated upregulation of VEGF and PEDF were observed in RPE from rats exposed to NT. Conclusions. NT increased the VEGF-to-PEDF ratio in the RPE through nAchR in vitro and in vivo. This alteration in the ratio may play a key role in the progression to wet AMD in passive smokers.

UR - http://www.scopus.com/inward/record.url?scp=79957746980&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=79957746980&partnerID=8YFLogxK

U2 - 10.1167/iovs.10-6254

DO - 10.1167/iovs.10-6254

M3 - Article

C2 - 21330654

AN - SCOPUS:79957746980

VL - 52

SP - 3842

EP - 3853

JO - Investigative Ophthalmology and Visual Science

JF - Investigative Ophthalmology and Visual Science

SN - 0146-0404

IS - 6

ER -