Flupirtine attenuates sodium nitroprusside-induced damage to retinal photoreceptors, in situ.
Flupirtine has been shown to function as a neuroprotectant and is presently used in man to treat a number of conditions. The aim of this study was to investigate the specific antioxidant properties of flupirtine in relation to oxidant-induced damage to retinal photoreceptors. Initial in vitro studies on brain membranes showed that flupirtine was approximately 20 times more potent than trolox (vitamin E analogue) and 8 times more potent than metipranolol at attenuating lipid peroxidation caused by the nitric oxide donor, sodium nitroprusside (SNP). Subsequent immunohistochemical studies revealed that following an intraocular injection of SNP, retinal photoreceptors are the only retinal cell types that appear to be clearly affected. This was supported by electroretinogram (ERG) recordings which showed both the a- and b-wave amplitudes to be significantly reduced. Western blotting techniques showed that SNP caused a significant decrease in photoreceptor-specific markers (RET-P1, rhodopsin kinase), an increase in cleaved caspase-3, Bcl-2, and cleaved PARP proteins that are associated with apoptosis and no change in the ganglion cell specific marker, neurofilament (NF-L). This was supported by RT-PCR data where rhodopsin (photoreceptor specific) mRNA was reduced while Thy-1 and NF-L (ganglion cell specific) mRNAs were unaffected. In addition SNP caused an elevation of glial cell response mRNAs primarily associated with Müller cells (GFAP, CNTF, bFGF) as well as caspase-3 and Bcl-2. Importantly, when flupirtine was co-injected, the effects to the retina caused by SNP on retinal proteins and mRNAs were in most cases significantly blunted. The conclusion reached from this study is that flupirtine is a powerful antioxidant and when injected into the eye with SNP attenuates the detrimental influence of SNP to retinal photoreceptors. Since oxidative stress has been implicated in retinal diseases like age-related macular degeneration (AMD) this study provides "proof of principle" for the idea that flupirtine may help individuals suffering from such retinal diseases.
Fawcett
R J
RJ
Nuffield Laboratory of Ophthalmology, Oxford University, Walton Street, Oxford OX2 6AW, UK.
Osborne
N N
NN
D000631:Aminopyridines / Q000378:metabolism*; D000700:Analgesics / Q000378:metabolism*; D000818:Animals; D000959:Antihypertensive Agents / Q000378:metabolism; D000975:Antioxidants / Q000378:metabolism; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002839:Chromans / Q000378:metabolism; D020934:Ciliary Neurotrophic Factor / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004596:Electroretinography; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism; D006801:Humans; D020287:In Situ Nick-End Labeling; D015227:Lipid Peroxidation; D014290:Metipranolol / Q000378:metabolism; D018696:Neuroprotective Agents / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000378:metabolism*; D009599:Nitroprusside / Q000378:metabolism*; D010786:Photoreceptor Cells / Q000166:cytology / Q000378:metabolism* / Q000473:pathology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D012243:Rhodopsin / Q000235:genetics / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism
D000631:Aminopyridines; D000700:Analgesics; D000959:Antihypertensive Agents; D000975:Antioxidants; D002839:Chromans; D020934:Ciliary Neurotrophic Factor; D005904:Glial Fibrillary Acidic Protein; D018696:Neuroprotective Agents; D020030:Nitric Oxide Donors; D019253:Proto-Oncogene Proteins c-bcl-2; D017392:Thiobarbituric Acid Reactive Substances; D016222:Fibroblast Growth Factor 2; D009599:Nitroprusside; D012243:Rhodopsin; D053148:Caspase 3; C034161:flupirtine; C010643:6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid; D014290:Metipranolol