| ObjectiveErythropoietin(EPO) as a acid glycoprotein containing acetylneuraminate hadbeen regarded as a body fluid factor concerning erythropoiesis and benefition of differentiation and proliferation. Recent research showed astrocyte secreted EPO by paracrine in the central nervous system(CNS), which can bind the receptor on the adjacent neuron membrane and produce protective effect. The possible mechanism might include inhibiting the neurotoxicity of glutaminic acid, increasing the interflow of calcium ion then inhibiting NO synthesis , preventing apoptosis,and alleviate peroxidation reaction by decreased oxygen-derived free radidical level.This objective was to investigate whether EPO has protective effect to this injury and the possible mechanism of EPO by detecting MDA and SOD level in brain tissue of Mongolian gerbil after ischemic reperfusion. Material and Methods: 1 .Animal and groups:Animal: 40 healthy mature Mongolian gerbilGroups: All animals were divided into 5 groups randomly,8 in each group. (1) pseudooperation group:operation as same as control group occlusion of bloodflow.(2) control group: inject 2ml saline through abdominal cavity at the beginning of reperfusion(3) EPO1500IU/kg(epo 15) group(4) EPO3000IU/kg(epo30) group(5)EP06000IU/kg(epo60) group(3)(4)(5)groups :inject 2ml saline solution containing EPO 1500IU/kg , 3000IU/kg ,6000IU/kg through abdominal cavity at the beginning of reperfusion respectively.2. reagent and drugs:MDA Test Kit, SOD Test Kit, recombinant human erythropoietin(rHu-EPO), et al.3. instruments: Mechanical horrogenizer ,low temperature centrifuger, UV2755B-t-ype ultraviolet-visible spectrophotometer.4.methods:4.1 Ischemic-reperfusion animal model:The method was to anesthetize Mongolian gerbil with Chloral Hydrate by abdominal cavity injection, segregate bilateral common carotid artery and clip two vessel with micro-vascular clamp after the animal waked up, occlude the bloodflow for 10m-in, then take the clamp away and suture Incisions.4.2 The criteria of model:the animals immediately appeared tachypnea ,cramp,and lost consiousness within 5~60s as soon as we occulded the bilateral common carotid artery.When the clamps were taken away,it came round in 3min.4.3 Four hours after ischemic-reperfusion, take out of the brain quickly,separate the part above the tentorium on a ice board, then make 10% tissue homogenate, centrifu-gation under 4h circumstance,measure the content of MDA and SOD in the supernate.5.Statistical methods: all data were processed through SAS6.12 for windows andSPSS11.0;the method include one-way ANOVA analysis,Least-Significant Differeceand Bivariate Correlate analysis.Results1.The study made the model succesfully with high stability (control group 72.7% vs EPO group 82.8%).2.The change of MDA in tissue of brainAfter ischemic-reperfusion, the MDA level increased significantly than pseudo-ooperation group(P<0.05). With the intervention of recombinant human erythropoiet-in(rHu-EPO),the MDA level decreased than control group(P<0.05).The MDA level d-ecreased with the increase of dosage EPO in each group,there were statistic significance among the groups(P<0.05). The dosages of rHu-Epo in three groups have negativ-e correlation to the MDA level in the tissue of brain (r=-0.851) ,indicating the dose-effect relation between the two variances. 3. The change of SOD in tissue of brainAfter ischemic-reperfusion, the SOD vitality decreased(P<0.05),The SOD vitality of each rHu-Epo treatment group increased significantly than control group (P<0.05).The SOD in epo60 group increased slightly than epo30 group (P>0.05).The SOD in other groups increased with the dose increasing of rHu-EPO(P<0.05). The dosages of rHu-Epo in three groups have positive correlation to the SOD vitality in the tissue of brain (r=0.813) .indicating the dose-effect relation between the two variances. Conclusion1. The oxidation injury and the disequilibrium... |
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