Neuroprotection by melatonin against ischemic neuronal injury associated with modulation of DNA damage and repair in the rat following a transient cerebral ischemia.
In the present study, double fluorescence staining combined with confocal laser scanning microscopy analysis were used to examine the effects of melatonin on ischemia-induced neuronal DNA strand breaks and its possible mechanisms in a transient middle cerebral artery (MCA) occlusion model. Results showed that melatonin dose-dependently reduced infarct areas and decreased both DNA double and single strand breaks (DSB and SSB) and enhanced cell viability in the peri-ischemic brain regions. Furthermore, Bcl-2 induction in the ischemic brain was further enhanced by melatonin treatment. Double staining analysis indicated that the cells costained for Bcl-2 and TdT-mediated-deoxyuridine triphosphate (dUTP) nick-end labeling (TUNEL), a DSB marker, displayed a relative regular morphology compared with the cells only stained with TUNEL. Transient ischemia induced an expression of excision repair cross-complementing factor 6 (ERCC6) mRNA, a gene essential for the preferential repair of nuclear excision repair, in the injured neurons. Double labeling showed that ERCC6 only co-localized with proliferating cell nuclear antigen (PCNA), a member of the nuclear excision repair complex, but not with TUNEL. Melatonin further and statistical significantly up-regulated ERCC6 mRNA expression in the peri-ischemic region of rat brains. The results suggest that neuroprotection by melatonin against ischemic injury may be related to modulation of apoptosis and DNA repair capacity.
National Key Laboratory of Medical Neurobiology, Medical Center of Fudan University, Shanghai, China. fysun@shmu.edu.cn
Feng-Yan
FY
Sun
Xiang
X
Lin
Li-Zhen
LZ
Mao
Wei-Hong
WH
Ge
Ling-Mei
LM
Zhang
Ya-Lin
YL
Huang
Jin
J
Gu
D000818:Animals; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology; D002545:Brain Ischemia / Q000473:pathology / Q000517:prevention & control*; D002536:Cerebral Arteries; D004247:DNA / Q000187:drug effects; D004249:DNA Damage / Q000187:drug effects*; D004265:DNA Helicases / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D004260:DNA Repair / Q000187:drug effects*; D004277:DNA, Single-Stranded / Q000187:drug effects; D004195:Disease Models, Animal; D020287:In Situ Nick-End Labeling; D008297:Male; D008550:Melatonin / Q000494:pharmacology*; D018613:Microscopy, Confocal; D009474:Neurons / Q000187:drug effects / Q000473:pathology*; D018696:Neuroprotective Agents / Q000494:pharmacology*; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D004277:DNA, Single-Stranded; D018696:Neuroprotective Agents; D019253:Proto-Oncogene Proteins c-bcl-2; D004247:DNA; D004265:DNA Helicases; D008550:Melatonin