A Experimental Study On Neuroprotective Effects Of Granulocyte Colony-Stimulating Factor In Cerebral Ischemia/Reperfusion Injury

Cerebrovascular diseases are a sort of diseases that harm humans health seriously.They have high mortality and disability and bring huge load to society. Cerebrovascular disease is the second cause of death worldwide.In our country,the number of Cerebrovascular disease is the first cause of death every city.However,no effective treatment currently exists.Thus,the basic and clinical study on Cerebrovascular disease are the main task of medical study.It was considered before that the neurous of adult was final stage and could not regenerate again when injuried.But more and more evidence indicate that the brain has the function of self-repairing.In adult brain,there have nerve stem cells(NSCs) in Subventricular zone(SVZ) and dentate gyrus hippocampus Subgranular zone(SGZ).These NSCs have the potentialty of self-renewing and multidifferentiation.Cerebral ischemia can lead endogenous neurogenesis,but the amount is few.Its capacity of self-repairing is limited.Although NSCs transplantation bring us hope to treat ischemic injury,but a lot of factor such as ethics, origin,transplantation injury,reject reaction limited its clinical apply.Mobilize bone marrow stem cells is a new method of cell transplantation,the stem cells which in bone marrow enter into peripheral blood making use of cell factor and arrive injuried organization.Granulocyte Colony-Stimulating Factor(G-CSF) is a strong mobilization reagent to bone marrow stem cells.It can increase the amount of stem cells in peripheral blood ,which move to cerebral ischemia area,differentiate to nerve cells in cerebral iechemia situation,repair injuried brain and protect nerve cells from damage. PartⅠAntiapoptosis effects of recombinant human granulocyte colony-stimulating factor in focal cerebral ischemia ratsObjective To study Recombinant Human Granulocyte Colony-Stimulating Factor(rhG-CSF) has antiapoptosis effects in focal cerebral ischemia Rats.Methods A model of focal cerebral ischemia /reperfusion in rats was performed with the intraluminal filament occlusion.To investigate the effects of rhG-CSF on neurological severity score and infarct volumes. Reverse transcriptional polymerase chain reaction(RT-PCR) was used to detect Caspase-3 mRNA expression in cerebral ischemia zone.Flow cytometry(FCM) was used to detect the rate of apoptosis in cerebral ischemia zone.Results Compare with contrast group, rhG-CSF significantly reduced the neurological severity scores and infarct volumes. rhG-CSF decreased the expression of Caspase-3 mRNA and the rate of apoptosis in cerebral ischemia zone.Conclusion rhG-CSF may reduced infarct volumes , decreased Caspase-3 mRNA expression and the rate of apoptosis in cerebral ischemia zone, rhG-CSF has antiapoptsis effects in focal cerebral ischemia/reperfusion injury.PartⅡAntiinflammatory effects of recombinant human granulocyte colony-stimulating factor in focal cerebral ischemia ratsObjective To study Recombinant Human Granulocyte Colony-Stimulating Factor(rhG-CSF) has antiinflammatory effects in focal cerebral ischemia Rats.Methods A model of focal cerebral ischemia /reperfusion in rats was performed with the intraluminal filament occlusion. Using immunohistochemistry to measure the expression of myeloperoxidase(MPO).Reverse transcriptional polymerase chain reaction(RT-PCR) was used to detect IL-1βmRNA和TNF-αmRNA expression in cerebral ischemia zone.Results Compare with contrast group, the expression of MPO was no deference between two group.But rhG-CSF decreased the expression of IL-1βmRNA和TNF-αmRNA .Conclusion rhG-CSF may decrease IL-1βmRNA和TNF-αmRNA expression and has antiinflammatory effects in focal cerebral ischemia/reperfusion injury.PartⅢInducing neuronal differentiation and angiogenesis effects of recombinant human granulocyte colony-stimulating factor in focal cerebral ischemia ratsObjective To investigate Recombinant Human Granulocyte Colony-Stimulating Factor(rhG-CSF) induce neuronal differentiation and angiogenesis in focal cerebral ischemia Rats.Methods A model of focal cerebral ischemia /reperfusion in rats was performed with the intraluminal filament occlusion.To evaluate neurological severity score of rhG-CSF treated group and contrast group in different reperfusion time,and using immunohistochemistry to measure the expression of neuroepithelial stem protein(Nestin),5-bromodeoxyuridine(Brdu),glial fibrilary acidic protein(GFAP),Neuronal Nuclei(NeuN),von Willebrand factor(vWF).Results Compare with contrast group, rhG-CSF significantly reduced the neurological severity scores and increased the expression of Nestin,Brdu, GFAP, NeuN, vWF in peri-infarcted zones. Conclusion the rhG-CSF treatment can induce neuronal differentiation and angiogenesis, can enhance the repairment of neuronal cells and have neuroprotective effects.