Oxidative DNA injury after experimental intracerebral hemorrhage.
Our previous studies have demonstrated that DNA injury occurs in the brain after intracerebral hemorrhage (ICH). DNA damage can result from at least two pathways, either endonuclease-mediated DNA fragmentation or oxidative injury. The present study investigated the occurrence of the latter after ICH and the role of iron in such injury. Male Sprague-Dawley rats received an infusion of autologous whole blood or ferrous iron into the right basal ganglia. Control rats just had a needle insertion (sham). The rats were sacrificed 1, 3, or 7 days later. 8-Hydroxyl-2'-deoxyguanosine (8-OHdG) was analyzed by immunohistochemistry while the number of apurnic/apyrimidinic abasic sites (AP sites) was also quantified. 8-OHdG and AP sites are two hallmarks of DNA oxidation. Dinitrophenyl (DNP) was measured by Western blotting to compare the time course of protein oxidative damage to that of DNA. DNA repair Ku proteins were measured by Western blot analysis. DNA damage was also examined using DNA polymerase I-mediated biotin-dATP nick translation (PANT) labeling. An increase of 8-OHdG, AP sites and DNP levels and a decrease of Ku levels were observed. Abundant PANT-positive cells were also observed in the perihematomal area 3 days after ICH. In addition, intracerebral infusion of iron increased brain DNP levels and resulted in DNA injury. These results suggest that oxidative stress contributes to DNA damage and brain injury after ICH. Reducing DNA oxidative damage (for example, through iron chelation) may be a therapeutic target for ICH.
Nakamura
Takehiro
T
Department of Neurosurgery, University of Michigan, 5550 kresge I, Ann Arbor, MI 48109-0532, USA.
Keep
Richard F
RF
Hua
Ya
Y
Hoff
Julian T
JT
Xi
Guohua
G
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D001479:Basal Ganglia / Q000187:drug effects / Q000378:metabolism*; D002543:Cerebral Hemorrhage / Q000188:drug therapy / Q000378:metabolism*; D004249:DNA Damage / Q000187:drug effects / Q000502:physiology*; D045643:DNA Repair Enzymes / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000378:metabolism; D003849:Deoxyguanosine / Q000031:analogs & derivatives / Q000378:metabolism; D004195:Disease Models, Animal; D007501:Iron / Q000008:administration & dosage*; D008297:Male; D008845:Microinjections; D018696:Neuroprotective Agents / Q000008:administration & dosage*; D018384:Oxidative Stress / Q000187:drug effects / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley
D004277:DNA, Single-Stranded; D018696:Neuroprotective Agents; D000080242:8-Hydroxy-2'-Deoxyguanosine; D007501:Iron; D045643:DNA Repair Enzymes; D003849:Deoxyguanosine