A Research Of Therapeutical Effect And Mechanism On G-CSF To Ischemia-reperfusion Injury In Rats Brain

Cerebral vascular disease(CVB)is one of three most threaten disease to people's lives and health. As so far, there is no effective method to cure the death of neuron. It's important to protect the ischemic neuron and make neuron regenerate. It provide a new method to cure the cerebral infarction by studying the protective effect and mechanism of granulocyte colony-stimulating factor(G-CSF) to ischemia-reperfusion injury in rats brain. We use proteomics to research difference protein of Ischemia-reperfusion Injury in Rats Brain after curing by G-CSF, which further indicate the therapeutical effect mechanism of G-CSF.At first, we testify the protective effect to Ischemia-reperfusion Injury in Rats Brain. The transient middle cerebral artery occlusion models were applied and reperfused after 2 hours. G-CSF (50ug/kg/d) is injected subcutaneously on rats'backs for successive 5 days. Rats was executed and decapitated receptively after 24h,7d and 14d after reperfusion to get the brain. The results of TTC stain show that and there is cerebral infarction in both brain of ischemia-reperfusion injured rats (I/R group) and the rats cured by G-CSF (G group). There is significant difference between the I/R group and G group, which testify the therapeutic effect of G-CSF. The results of HE stain confirms the results above again.Secondly, immunohistochemical method and TUNEL stain are used to test Bcl-2,Caspase-3,IL-1β,NF-κBp65 and apoptosis. Results: It is hardly to see positive cells of each kind and apoptotic cells in N and S group. All kinds of positive cells and apoptotic cells are seen in I/R group, and there is significant difference between I/R and S group. All kinds of positive cells and apoptotic cells are significantly reduced in G group by contrast to I/R group. At last, to research difference protein of ischemia-reperfusion injury in rats brain after curing by G-CSF and to screen relative protein of neuron protective effect by proteomics technology. Result: Compared with I/R group, the S group gained 56 differential protein spots, 17 spots expressed lowly, and 39 spots high, identified 19 protein spots,found out the relative cerebral ischemic proteins. Compared with I/R group, the G group gained 15 differential protein spots,3 spots expressed lowly,and 13 spots high,identified 6 protein spots, It's found out the relative protective proteins of G-CSF such as GFAP, endomucin and DRP-2 etc.Conclusion: In this experiment, it is proved that the neuron protective effect of G-CSF exists and G-CSF has a anti-apoptotic and anti-inflammatory effect to ischemia-reperfusion Injury in Rats Brain. By using proteomics, we know some proteins about cerebral ischemia , which further explore the mechanism of therapeutic effect of G-CSF .