Infliximab reduces Zaprinast-induced retinal degeneration in cultures of porcine retinaReport as inadecuate




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Journal of Neuroinflammation

, 11:172

First Online: 10 October 2014Received: 07 June 2014Accepted: 25 September 2014

Abstract

BackgroundcGMP-degrading phosphodiesterase 6 PDE6 mutations cause around 4 to 5% of retinitis pigmentosa RP, a rare form of retinal dystrophy. Growing evidence suggests that inflammation is involved in the progression of RP. The aims of this study were to corroborate the presence of high TNFΑ concentration in the eyes of RP patients and to evaluate whether the blockade of TNFΑ with Infliximab, a monoclonal anti-TNFΑ antibody, prevented retinal degeneration induced by PDE6 inhibition in cultures of porcine retina.

MethodsAqueous humor from 30 patients with RP and 13 healthy controls were used to quantify the inflammatory mediators IL-6, TNFΑ, IL-1Β, IL-10 by a multiplex enzyme-linked immunosorbent assay ELISA system. Retinal explants from pig were exposed to Zaprinast, a PDE6 inhibitor, for 24 hours in the absence or the presence of Infliximab. Cell death was evaluated by TUNEL assay. The number and distribution of caspase-3 positive cells, indirect polyADPribose polymerase PARP activation and glial fibrillary acidic protein GFAP content were visualized by immunolabeling. Antioxidant total capacity, nitrites and thiobarbituric acid reactive substances TBARS formation were determined to evaluate antioxidant-oxidant status.

ResultsIL-6 and TNFΑ concentrations were higher in the aqueous humor of RP patients than in controls. Infliximab prevented retinal degeneration, as judging by the reduced presence of TUNEL-positive cells, the reduction of caspase-3 activation and also reduction of glial activation, in an ex vivo model of porcine retina. Additionally, Infliximab partially reduced oxidative stress in retinal explants exposed to Zaprinast.

ConclusionsInflammatory mediators IL-6 and TNFΑ were elevated in the aqueous humor of RP patients corroborating previous studies suggesting sustained chronic inflammation. Our study suggests that TNFΑ is playing an important role in cell death in an ex vivo model of retinal degeneration by activating different cell pathways at different cell layers of the retina that should be further studied.

KeywordsRetinal degeneration Inflammation Infliximab Oxidative stress TNFΑ PolyADP-ribose caspase-3 Retinitis pigmentosa Photoreceptor death AbbreviationsAIFapoptosis inducing factor

AMDage related macular degeneration

ANCOVAanalysis of variance

auarbitrary units

BCVAbest-corrected visual acuity

BCAbicinchoninic acid

BSAbovine serum albumin

cGMPcyclic guanosine monophosphate

DMSOdimethyl sulfoxide

ELISAenzyme-linked immunosorbent assay

GCLganglion nuclear layer

GFAPglial fibrillary acidic protein

ILinterleukin

INLinner nuclear layer

iNOXintracellular nitrite

IRischemic-reperfusion

MANCOVAmultivariate analysis of covariance

MDAmalonyldialdehyde

NOnitric oxide

NOXintracellular nitrates and nitrites

ONLouter nuclear layer

PARpolyADP-ribose

PARPpolyADP-ribose polymerase

PBSphosphate-buffered saline

PDE6phosphodiesterase 6

RPretinitis pigmentosa

TACtotal antioxidant capacity

TBAthiobarbituric acid

TBARSthiobarbituric acid reactive substances

TLRsToll-like receptors

TNFΑtumor necrosis factor alpha

TUNELterminal deoxynucleotidil transferase dUTP nick and labeling

VFvisual field

Electronic supplementary materialThe online version of this article doi:10.1186-s12974-014-0172-9 contains supplementary material, which is available to authorized users.

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Author: Cristina Martínez-Fernández de la Cámara - Lorena Olivares-González - David Hervás - David Salom - José M Millán - Re

Source: https://link.springer.com/







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