Involvement of oxidative and nitrosative stress in promoting retinal vasculitis in patients with Eales' disease.
OBJECTIVE
Eales' disease (ED) is an idiopathic retinal vasculitis condition, which affects retina of young adult males. The histopathological hallmark in ED is the adhesion of leukocytes to the endothelium and the infiltration of these cells into the retinal parenchyma. Phagocyte generated free radicals have been implicated in mediating tissue damage associated with various inflammatory vasculopathies. In the present study, we have investigated the possible role of reactive oxygen species (ROS) and reactive nitrogen species (RNS) in causing retinal tissue damage in ED.


METHODS
35 patients with ED and 20 healthy control subjects were included in the study. Monocytes (MC) were separated from peripheral blood of the respective study participants. Inducible nitric oxide synthase (iNOS) protein expression was assessed using Western blot and 3 nitrotyrosine (3NTYR) by reversed phase high performance liquid chromatography (RP HPLC). Thiobarbituric acid reactive substances (TBARS) were determined by measuring malondialdehyde (MDA) formed. Superoxide dismutase (SOD) activity was assayed based on the ability of SOD to inhibit auto-oxidation of epinephrine. Iron, copper and zinc content were determined using Atomic Absorption Spectrophotometer. Immunolocalization of iNOS and 3NTYR was performed on the surgically excised epiretinal membranes (ERM) from patients with ED.


RESULTS
There was a significant increase in the expression of iNOS, as well as 3NTYR accumulation, diminished SOD activity, elevated lipid peroxides, iron, copper and decreased zinc content in the MC of patients with ED when compared with healthy control subjects. The elevated levels of ROS and RNS products correlated with diminished antioxidant status in patients with ED. Strong immunoreactivity for iNOS and 3NTYR was observed in inflammatory cells and endothelial cells in ERM obtained from patients with ED.


CONCLUSIONS
Our findings from this study clearly reveal the involvement of RNS and ROS in the development of retinal vasculitis in ED. Based on our present study and earlier studies we confirm the role of free radicals in mediating retinal tissue damage in ED. Hence we believe selective inhibition of iNOS or supplementation with antioxidants vitamin E and C might be beneficial in controlling retinal vasculitis in patients with ED.
Rajesh
Mohanraj
M
Biochemistry Research Department, Vision Research Foundation, Sankara Nethralaya, 600006 Chennai, India.
Sulochana
Konerirajapuram N
KN
Punitham
Ranganathan
R
Biswas
Jyotirmay
J
Lakshmi
Soundarajan
S
Ramakrishnan
Sivaramakrishnan
S
D000328:Adult; D015153:Blotting, Western; D002851:Chromatography, High Pressure Liquid; D003300:Copper / Q000032:analysis; D006801:Humans; D007501:Iron / Q000032:analysis; D008297:Male; D008315:Malondialdehyde / Q000032:analysis; D009000:Monocytes / Q000166:cytology / Q000378:metabolism; D009566:Nitrates / Q000737:chemistry / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000096:biosynthesis; D015538:Nitrosation; D010084:Oxidation-Reduction; D018384:Oxidative Stress / Q000502:physiology*; D031300:Retinal Vasculitis / Q000378:metabolism* / Q000503:physiopathology; D012171:Retinal Vessels / Q000473:pathology*; D013054:Spectrophotometry, Atomic / Q000379:methods; D013482:Superoxide Dismutase / Q000378:metabolism; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis; D015032:Zinc / Q000032:analysis
D009566:Nitrates; D017392:Thiobarbituric Acid Reactive Substances; C002744:3-nitrotyrosine; D014443:Tyrosine; D008315:Malondialdehyde; D003300:Copper; D007501:Iron; D019001:Nitric Oxide Synthase; D013482:Superoxide Dismutase; D015032:Zinc