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Effects of Erythropoietin on Expression of BrdU and GFAP in Brain of Neonatal Rat with Hypoxic-ischemic Brain Damage

Author: WangXuQin
Tutor: CaoYunTao
School: Zunyi Medical College,
Course: Pediatrics
Keywords: Hypoxic-ischemic brain damage rats newborn Erythropoietin 5-bromo-2-deoxyuridine Glial fibrillary acidic protein Morris water maze
CLC: R722.1
Type: Master's thesis
Year: 2011
Downloads: 70
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Abstract

Objective:In this study we investigate the effects of erythropoietin(EPO) on the ex-pression of 5-bromo-deoxyuridine(BrdU) in dentate gyrus(DG) region and glial fibri-llary acidic protein(GFAP) in CA1 region after hypoxic-ischemic brain damage(HIB D) in neonatal rats,to explore the neuroprotective effects of erythropoietin according to the regeneration of neurocytes and the improvement of learning and memory ca-pability by using the Morris water maze, in order to obtain the experimental evide-nces of the suitable therapeutic dose of the EPO for hypoxic-ischemic encephalopat-hy ofneonates.Methods:One hundred sixty neonatal Wistar rats were divided into five groups ran-domly:the group of the HIBD,the control group of normal sodium and the trial g-roup of EPOⅠ,Ⅱ,Ⅲ, each group were thirty-two, the HIBD models of neonatal Wistar rats were made by shearing right arteria carotis communis and then breathi-ng 8%O2+92%N2 for two hours. the trial groups of EPOⅠ、Ⅱ、Ⅲwere injected intraperitonally separate rhEPO 5u/g、10u/g、15u/g for fourteen days since operation, the control group of the HIBD was injected intraperitonally 0.5ml NS as a pair,The other forty rats were taken into the sham operation group. Before every rat wassac-rificed,5-bromodeoxyuridine(BrdU) was injected intraperitonally three times in order to label newly generated neurocytes exclude the rats for spatial cognitive capability by using the Morris water maze in each groups. The expressions of BrdU and GF AP in dentate gyrus(DG) region and the CA1 region were examined with immuno-histochemical staining and image quantitative analysis 14d,21d,28d after the oper-ation. Pathological change of brain was investigated with microscope. Each of five group’s spatial cognitive capability was evaluated by using the Morris water maze also.Results:1.the expression of 5-bromo-deoxyuridine(BrdU) in dentate gyrus(DG) regio-n:The number of BrdU-positive cells in hippocampal region was gradually decrees- ing accompany with the increasing age in each groups(P<0.05);The BrdU-positive c-ells were a significantly difference of the five groups(P<0.05), The expression of BrdU-positive cells of the EPO trial groups were higher than the control group,and the sham operation group was least compare with other groups (there is a significa-ntly difference in five groups at the 14th and 21th days after the operation, but no significantly difference at the 28th days after the operation);The number of BrdU-positive cells in dentate gyrus(DG) region had decreasing tendency accompany with the increase therapeutic dose of the EPO at all points in the trial groups,but there is no significantly difference(P>0.05).2. the expression of GFAP in CA1 region:The number of GFAP in CA1 region was gradually decreasing accompany with the inc-reasing days,there is a significantly difference(P<0.05);The expression of GFAP-posi-tive cells was the most in the trial group of EPO, second is the control group of HIBD,the sham operation group was least (there is a significantly difference in five groups at the 14th and 21th days after the operation, but no significantly difference at the 28th days after the operation);The number of GFAP-positive cells in CA1 re-gion had decreasing tendency accompany with the increase therapeutic dose of the EPO at all points in the trial groups,but there is no significantly difference(P>0.05). 3. Determination of Morris water maze:①Learning ability:The latency to escape (LE) was significantly difference in five groups(P<0.05), the sham group is the sh-orter compared with the EPOⅠ,Ⅱ,Ⅲtreated groups, the control group of HIBD is the longest.②Memory capacity:the swimming time in platform surrounding area NE was a significantly difference in five groups; there is a significantly difference in the sham operation group and the EPOⅠ,Ⅱ,Ⅲtreated group compared with the control group of HIBD(P<0.05), but there was no significant difference between the sham operation group and the EPOⅠ,Ⅱ,Ⅲtreated group(P>0.05). and also no sig-nificant difference in the EPOⅠ,the EPO II and the EPOIII groups too(P>0.05).Conclusions:1. Exogenous EPO could increases the expression of BrdU and GFAP in hippocampal region of neonatal rats with HIBD, it means that EPO promotes the regeneration of neurocytes damaged by hypoxia-ischemia and it may be involved in the regeneration of neurocytes and play an important role of neuroprotective effects in neonatal rats with HIBD;2. EPO early intervention can improve the ability of learning and memory in the neonatal rats with HIBD;3. Hypoxic-ischemic brain damage could improve the proliferation differentiation and regeneration of the endogenous neural stem cells in the neonatal rats with HIBD;4.The result show there is a decreasing tendency of the marks that represents the regeneration of neurocytes accompany with increased therapeutic dose of the EPO in the neonatal rats with HIBD,it could not improve the neuroprotective effects by increasing dose of EPO.

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