Erythropoietin prevents zinc accumulation and neuronal death after traumatic brain injury in rat hippocampus: in vitro and in vivo studies.
Erythropoietin (Epo) has been gaining great interest for its potential neuroprotective effect in various neurological insults. However, the molecular mechanism underlying how Epo exerts the function is not clear. Recent studies have indicated that Zn(2+) may have a key role in selective cell death in excitotoxicity after injury. In the present study, we studied the effect of recombinant human Epo (rhEpo) in zinc-induced neurotoxicity both in vitro and in vivo. Exposure of cultured hippocampal neurons to 200 muM ZnC1(2) for 20 min resulted in remarkable neuronal injury, revealed by assessing neuronal morphology. By measuring mitochondrial function using MTT assay, we found that application of rhEpo (0.1 U/ml) 24 h before zinc exposure resulted in a significant increase of neuronal survival (0.6007+/-0.2280 Epo group vs 0.2333+/-0.1249 in control group; n=4, p<0.01). Furthermore, we demonstrated that administration of rhEpo (5,000 IU/kg, intraperitoneal) 30 min after traumatic brain injury (TBI) in rats dramatically protected neuronal death indicated by ZP4 staining, a new zinc-specific fluorescent sensor which has been widely used to indicate neuronal damage after excitotoxic injury (n=5/group, p<0.05). Neuronal damage was also assessed by Fluoro-Jade B (FJB) staining, a highly specific fluorescent marker for the degenerating neurons. Consistent with ZP4 staining, we found the beneficial effects of rhEpo on neuronal survival in hippocampus after TBI (n=5/group, p<0.05). Our results suggest that rhEpo can significantly reduce the pathological Zn(2+) accumulation in rat hippocampus after TBI as well as zinc-induced cell death in cultured cells, which may potentially contribute to its neuronal protection after excitotoxic brain damage.
Zhu
Lin
L
Department of Neurosurgery, Jinling Hospital, Nanjing, Jiangsu Province, China.
Wang
Han-dong
HD
Yu
Xin-guang
XG
Jin
Wei
W
Qiao
Liang
L
Lu
Ting-jia
TJ
Hu
Ze-lan
ZL
Zhou
Jing
J
D000704:Analysis of Variance; D000818:Animals; D001930:Brain Injuries / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D002452:Cell Count; D016923:Cell Death / Q000187:drug effects*; D002478:Cells, Cultured; D002712:Chlorides / Q000633:toxicity; D004305:Dose-Response Relationship, Drug; D004921:Erythropoietin / Q000494:pharmacology / Q000627:therapeutic use*; D005452:Fluoresceins / Q000378:metabolism; D006624:Hippocampus / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D007150:Immunohistochemistry; D008297:Male; D009474:Neurons / Q000187:drug effects / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D011994:Recombinant Proteins; D015032:Zinc / Q000378:metabolism; D017967:Zinc Compounds / Q000633:toxicity
D002712:Chlorides; D005452:Fluoresceins; D011994:Recombinant Proteins; D017967:Zinc Compounds; C501988:Zinpyr-4; D004921:Erythropoietin; C016837:zinc chloride; D015032:Zinc