Iron-induced oxidative brain injury after experimental intracerebral hemorrhage
Part of the
Acta Neurochirurgica Supplementum
book series (NEUROCHIRURGICA, volume 96)
We investigated the occurrence of DNA damage in brain after intracerebral hemorrhage (ICH) and the role of iron in such injury.
Male Sprague-Dawley rats received an infusion of 100 µL autologous whole blood or 30 µL FeCl2 into the right basal ganglia and were sacrificed 1, 3, or 7 days later. 8-hydroxyl-2′-deoxyguanosine (8-OHdG) was analyzed by immunohistochemistry, while the number of apurinic/apyrimidinic abasic sites (AP sites) was also quantified. 8-OHdG and AP sites are two hallmarks of DNA oxidation. DNA damage was also examined using PANT and TUNEL labeling. Dinitrophenyl (DNP) was measured by Western blot to compare the time course of protein oxidative damage to that of DNA. DNA repair APE/Ref-1 and Ku-proteins were also measured by Western blot. Bipyridine, a ferrous iron chelator, was used to examine the role of iron in ICH-induced oxidative brain injury.
An increase in 8-OHdG, AP sites, and DNP levels, and a decrease in APE/Ref-1 and Ku levels were observed. Abundant PANT-positive cells were also observed in the perihematomal area 3 days after ICH. Bipyridine attenuated ICH-induced changes in PANT and DNP. These results suggest that iron-induced oxidation causes DNA damage in brain after ICH and that iron is a therapeutic target for ICH.
KeywordsIntracerebral hemorrhage iron oxidative DNA injury 8-OHdG AP sites DNP PANT APE/Ref-1 Ku-proteins brain edema
Bridges KR, Cudkowicz A (1984) Effect of iron chelators on the transferrin receptor in K562 cells. J Biol Chem 259: 12970–12977PubMedGoogle Scholar
Connor JR, Menzies SL, Burdo JR, Boyer PJ (2001) Iron and iron management proteins in neurobiology. Pediatr Neurol 25:118–129PubMedCrossRefGoogle Scholar
Graham SH, Chen J (2001) Programmed cell death in cerebral ischemia. J Cereb Blood Flow Metab 21: 99–109PubMedCrossRefGoogle Scholar
Ikeda Y, Ikeda K, Long DM (1989) Comparative study of different iron-chelating agents in cold-induced brain edema. Neurosurgery 24: 820–824PubMedCrossRefGoogle Scholar
Kasai H, Crain PF, Kuchino Y, Nishimura S, Ootsuyama A, Tanooka H (1986) Formation of 8-hydroxyguanine moiety in cellular DNA by agents producing oxygen radicals and evidence for its repair. Carcinogenesis 7: 1849–1851PubMedGoogle Scholar
Kim GW, Noshita N, Sugawara T, Chan PH (2001) Early decrease in DNA repair proteins, Ku70 and Ku86, and subsequent DNA fragmentation after transient focal cerebral ischemia in mice. Stroke 32: 1401–1407PubMedGoogle Scholar
Lewen A, Sugawara T, Gasche Y, Fujimura M, Chan PH (2001) Oxidative cellular damage and the reduction of APE/Ref-1 expression after experimental traumatic brain injury. Neurobiol Dis 8: 380–390PubMedCrossRefGoogle Scholar
Nagasawa T, Hatayama T, Watanabe Y, Tanaka M, Niisato Y, Kitts DD (1997) Free radical-mediated effects on skeletal muscle protein in rats treated with Fe-nitrilotriacetate. Biochem Biophys Res Commun 231: 37–41PubMedCrossRefGoogle Scholar
Nakamura T, Keep RF, Hua Y, Schallert T, Hoff JT, Xi G (2004) Deferoxamine-induced attenuation of brain edema and neurological deficits in a rat model of intracerebral hemorrhage. J Neurosurg 100: 672–678PubMedCrossRefGoogle Scholar
Shacter E, Williams JA, Lim M, Levine RL (1994) Differential susceptibility of plasma proteins to oxidative modification: examination by Western blot immunoassay. Free Radic Biol Med 17: 429–437PubMedCrossRefGoogle Scholar
Siesjo BK, Agardh CD, Bengtsson F (1989) Free radicals and brain damage. Cerebrovasc Brain Metab Rev 1: 165–211PubMedGoogle Scholar
Wu J, Hua Y, Keep RF, Schallert T, Hoff JT, Xi G (2002) Oxidative brain injury from extravasated erythrocytes after intracerebral hemorrhage. Brain Res 953: 45–52PubMedCrossRefGoogle Scholar
Zhang Z, Wei T, Hou J, Li G, Yu S, Xin W (2003) Iron-induced oxidative damage and apoptosis in cerebellar granule cells: attenuation by tetramethylpyrazine and ferulic acid. Eur J Pharmacol 467: 41–47PubMedCrossRefGoogle Scholar