| Cerebral arterial thrombosis is one of the leading causes of unnatural death and disability. Satisfactory results have been acquired from the transplantation of bone marrow stromal cells (BMSCs) in the treatment of experimental cerebral ischemia. But it is still a complicated manipulation with high chance of infection and possibility of missing the time window for therapy. Recombinant human granulocyte colony-stimulating factor (rhG-CSF) as a mobilizing agent for bone marrow stem cells can mobilize both hematopoietic stem cells and BMSCs into blood circulation. RhG-CSF has been proved to be of significant benefit in the treatment of cardiac and cerebral ischemia. Bone marrow stromal cells have the ability of continuously secreting fibronectin and brain-derived neurotrophic factor (BDNF). In the present study, we observed the influence of rhG-CSF on the neurological deficits in focal cerebral ischemic rats and the expression of CD34 (specific marker for hematopoietic stem cells), fibronectin, BDNF and Bcl-2 in the ischemic brain tissue. We employed hippocampal slice cultures treated with oxygen and glucose deprivation (OGD) followed by oxygen-glucose re-supply to mimic the ischemia-reperfusion injury in experimental animals. In order to further explore the benefit of rhG-CSF on ischemic neurons, we observed the direct influence of fibronectin on OGD hippocampal slices and investigated the influences of OGD hippocampal slices on the differentiation of BMSCs into cells of nerve tissue without direct contact with each other.Part â… The effect of rhG-CSF on the expression of CD34, fibronectin, BDNF and Bcl-2 in the ischemic brain tissueInjection of 10μg/kg.d rhG-CSF for 5 days was given subcutaneously to rats undergoing 2 hours of middle cerebral artery occlusion (MCAO) at the 3rd hour after reperfusion. Equal volum of saline was given to control group. Each group was subdivided into 7, 14, 21d group. Neurological Severity Scores (NSS) tests were performed at the first day after reperfusion and at the time just before sacrifice. The expression of CD34, fibronectin, BDNF and Bcl-2 were observed by immunohistochemistry. To visualize the cellular colocalization of Brdu and fibronectin, glial fibrillary acidic protein (GFAP) and fibronectin, nestin and BDNF, Brdu and BDNF, double fluorescent staining was used.The results showed that: NSS of the rhG-CSF treated groups at 7th day, 14th day and 21st day after reperfusion were significantly lower than that of the control groups (P<0.05). The numbers of CD34 immunopositive cells in the marginal zone of the infarction in the rhG-CSF treated groups at 7th, 14th and 21st day after reperfusion were 25.95±4.30,16.65±2.85,9.32±1.30 respectively, those in the control groups were 17.42±3.86,10.95±1.80,6.40±1.04. The numbers of CD34 immunopositive cells in the rhG-CSF treated groups were statistically higher than those of the control groups(P<0.05). The numbers of fibronectin immunopositive cells in the marginal zone of the infarction in the rhG-CSF treated groups at 7th, 14th day after reperfusion were 20.90±5.11,17.83±3.45 respectively, those in the control groups at the same time points were 14.07±3.23,11.83±4.12. The numbers of fibronectin immunopositive cells in the rhG-CSF treated groups were significantly higher than those of the control groups(P<0.05). No fibronectin immunopositive cells were observed in the control groups at 21st day after reperfusion, but there were still a few fibronectin immunopositive cells in the marginal zone of the rhG-CSF treated groups at this time point. The numbers of BDNF immunopositive cells in the marginal zone of the infarction in the rhG-CSF treated groups at 7th, 14th and 21st day after reperfusion were 25.10±5.70, 16.78±3.75, 12.13±3.67 respectively, those of the control groups were 18.20±4.50, 11.70±2.93, 6.92±2.53. The numbers of the rhG-CSF treated groups were statistically higher than those of the control groups(P<0.05). The number of Bcl-2 immunopositive cells in the marginal zone of the infarction of...
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