Neuroprotection Effect Of Combined Application Of Erythropoietin And Granulocyte Colony-stimulating Factor In The Rat Cerebral Ischemia-reperfusion Injury

Objective:To research the neuroprotection effect of combined application of erythropoietin and granulocyte colony-stimulating factor in the rat cerebral ischemia-reperfusion injury.Methods:120 rats were randomly divided into four groups (Sham operation control group, reperfusion IR, reperfusion IR+G-CSF, reperfusion IR+G-CSF+EPO).Each group has 30 rats. Each respectively was divided one, three, seven days,namely three subgroups. Each subgroup has ten rats. The rat model of focal cerebral ischemia reperfusion was established by improved Zea Longa. Based on single-blind method, Bederson standard for evaluation is employed to determine the success of model making. Observe the neurological symptoms after the treatment of combined application of erythropoietin and granulocyte colony-stimulating factor in the rats cerebral ischemia-reperfusion.With HE staining,observe brain tissue pathological form change. Compare rats'death rate of each group, brain obstruction area, and routine blood changes. Compare apoptotic cells quantity of each group with TUNEL staining. Immunohistochemistry detect the expression of caspase-3.Results:1 Sham operation group showed no neural deficiency symptoms. Reperfusion IR group showed neurological symptoms of varying degrees. Observation indicates, reperfusion IR+G-CSF and reperfusion IR+G-CSF+EPO are able to improve neural deficiency symptoms of cerebral ischemia-reperfusion injury to varying degrees.2 HE staining showed that the neurons morphological structure of sham group was normal. In reperfusion IR group, ischemia side organization disappeared or was abnormal remarkablely. Cell number was reduced and cell arrangement was disordered. Reperfusion IR+G-CSF and reperfusion IR+G-CSF+EPO are able to prevent a reduction in brain tissue necrosis. Under the light microscope, results of prevention group was slightly worse.3 Cerebral infarction area:Obvious focus of infarct appeared when reperfuse on the first day, and mainly located in the cerebral cortex. Reperfuse on the third day. cerebral infarction area was the largest and it was reduced when reperfuse on the seventh day. Reperfuse one more day, there was no difference between the treatment groups and model groups. When reperfuse three,seven days, compared with model groups, infarct size of each treatment group decreased and infarct size of combined treatment groups decreased evidently.4 Death rate:Both the single treatment groups could cut the death rate. Compated with the single treatement groups, combined treatment groups results slightly worse.5 Blut-routine-untersuchung:There was no differences in HB,RBC,PLT at different time points in each groups. Reperfused one more day, the number of WBC increased. On the seventh day, it increased gradually. But there was no disctinct difference between model groups and the two treatment groups(p>0.05). Drug dose used in this experiment had no influence on animal blood index in a short time and it was safe.6 TUNEL detection:On the third day, the number of apoptotic cells peaked. Between model groups and treatment groups, on the first day the number of Immune positive cells was no significant difference. While on the third day and seventh day, the number of Immune positive cells in the two treatment groups was smaller than model groups. Compared with G-CSF groups, on the third day and seventh day the number of positive cells was significantly reduced.7 Expression of caspase-3:The expression of positive cells in the two treatment groups reduced. Compared with G-CSF groups, the number of positive groups in the combined treatment groups was significantly reduced.Conclusions:This experiment was a success in the rats model of cerebral ischemia reperfusion. The combined treatment of G-CSF and EPO obviously improved neurological symptoms in the rats of cerebral ischemia reperfusion, reduced infarction aera and rats death rate, and reduced brain ultrastructure damage. Both G-CSF and the combination of G-CSF and EPO could reduce the rat number of apoptosis cells and the latter had more power. The combination of G-CSF and EPO played together as a antiapoptotic role through the reduction of expression of caspase-3. Drug dose used in the combination treatment of G-CSF and EPO was safe.