Wydanie 2/2015
str. 18

Zastosowanie testu kometowego w celu oceny poziomu uszkodzeń oksydacyjnych DNA u chorych na jaskrę pierwotną otwartego kąta

Endonuclease Modified Comet Assay for Oxidative DNA Damage in Primary Open Angle Glaucoma Patients

Katarzyna Szymanek1, Jacek P. Szaflik1, Jerzy Szaflik1, Karolina Przybyłowska-Sygut2, Ireneusz Majsterek2, Patrycja Banaś-Leżańska2

1Katedra i Klinika Okulistyki II Wydziału Lekarskiego Uniwersytetu Medycznego w Warszawie
Kierownik: prof. dr hab. n. med. Jacek P. Szaflik
2 Zakład Chemii i Biochemii Klinicznej Międzywydziałowej Katedry Chemii i Biochemii Medycznej
Uniwersytetu Medycznego w Łodzi
Kierownik: prof. dr hab. n. med. Ireneusz Majsterek


Summary: Purpose:The aim of this study was to determine the amount of single-stranded DNA breaks in lymphocytes of patients with primary open angle glaucoma.
Material and methods:The material used for the study were human peripheral blood lymphocytes. Study group consisted of 5 patients with primary open angle glaucoma and 5 patients without signs of disease, matched for age. To assess the level of DNA damage and the effectiveness of the repair process used version of the alkaline comet assay of enzymes Fpg and endonuclease III (Endo III).
Results: Lymphocytes from glaucoma patients had significantly higher amount of DNA strand breaks observed in the comet tail, compared to the control group. Cells were isolated from the blood of patients with primary open angle glaucoma were more susceptible to oxidative DNA damage induced by enzymes Fpg and Nth than in the control group. In glaucoma patients, a statistically higher level of DNA damage than the control group (p <0.05). Percentage was significantly reduced oxidative damage repair glaucoma group compared to the control group.
Conclusions: Glaucoma patients exhibit significantly higher levels of DNA damage compared with healthy subjects. Oxidative stress showed a pathogenic role in primary open angle glaucoma. Evaluation of DNA damage can enrich our understanding of the pathology of eye diseases, including glaucoma, open angle and ultimately lead to finding new therapeutic directions.

Słowa kluczowe: jaskra, oksydacyjne uszkodzenia DNA, enzymy antyoksydacyjne, test kometowy.

Keywords: glaucoma, oxidative DNA damage, antioxidant enzymes, comet assay.


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