| Objective: Recently with economic development, population aging, the incidence of cerebral infarction is increasing and showing an upward trend year after year. It is a serious threat to human health. So it has been a hot research that how to restore neuronal function effectively. In recent years, with the biological characteristics of stem cell research and stem cell technology, the rapid development of the application of stem cell biology involved in almost every area, including of cerebrovascular disease. As an improved Zea-longa-line intraluminal occlusion of middle cerebral artery infarction model, which of SD rats were injected bone marrow stromal cells through the tail vein and intraperitoneal injection of granulocyte colony stimulating factor and compared both kinds of methods of treatment of cerebral infarction in rats. We studied neurological function and infarction after ischemia in rats, as well as the number of cell survival, migration, distribution of transplantation and stem cell mobilization. We studied the possible mechanisms of neuronal repair as well.Methods:①Animals: Adult healthy male Sprague Dawley (SD) rats of closed colony and clean grade were purchased from Laboratory Animal Center of Hebei Medical University. The treatment of animals in the experimental process was in accordance with the criteria of Animal Ethics.②Experimental Methods: 100 normal adult SD rats, were ischemia by Zea-Longa-line intraluminal middle cerebral artery occlusion model (MCAO) for 2 hours and unpluged a length of Nylons thread for blood flow reperfusion. Application neuronal assessment score after rats regained consciousness with reference to Longa method for five scoring. The model Rating 1 ~ 3 were successfully choozen. Select the successful model into the group. All rats were respectively given G-CSF10μg/kg intraperitoneally and were injected 1ml BMSCs via tail vein (containing about 1×106 bone marrow stromal cells) in 24 hours .Control groups were in the same way to give the same amount of PBS. Normal group received no treatment.③Experimental Evaluation: Intraperitoneal injection of G-CSF and intravenous injection of BMSCs in 7 day and 14 day, respectively. Morris water maze test was used to evaluate cognitive function in the same time in 7 day and 14 day. 6 rats of group of G-CSF and BMSCs were continuous perfusional paraffin section and immunohistochemically stained to detect infarctional area and BrdU-positive cells.Results:①MCAO rats changed in motor function: Swimming speed of BMSCs group in 7 day and 14 day after transplantation were with statistical significance compared with PBS groups in 7day and in14 day (P<0.05), and were higher than the PBS group. Swimming speed was statistically significant different between G-CSF groups in 7 day and in 14 day after intraperitoneal injection with the PBS group in 7day and in 14 day (P<0.05), and were both higher than the PBS group.There are significant differences in swimming speed of G-CSF group and BMSCs group in7 day and in 14 day (P<0.05), and G-CSF group was higher than BMSCs group .②Cognitive function change: Time of the search platform of groups of BMSCs and G-CSF were statistically significantly different compared with the Normal group and PBS group in 7 day (P<0.05), and were shorter than those in both latter groups. Time of the search platform of groups of BMSCs and G-CSF were statistically significantly different compared with the Normal group and PBS group in 14 day (P<0.05) and shorter than those in the control group. There were no statistically significant difference in BMSCs group and the PBS group in 14 day.③The percentage of infarction area: The percentage of infarction area of BMSCs group and G-CSF group (17.7±2.4%), (11.2±3.2%) were statistically significant difference with the PBS group (32.5±2.6%) (P<0.05) and were smaller than PBS group in 7 day. The percentage of infarction area of G-CSF group (7.0±2.1) and BMSCs group, PBS group (11.1±2.7%) (13.3±2.4%) were statistically significantly different in 14 day (P<0.05), and lower than both latter groups. BMSCs group and PBS group were no statistical difference in 14 day.④BrdU positive cells: The numbers of BrdU-positive cells of G-CSF group in 7 day and 14 day (174±32), (140±24) were statistically significantly different respectively with BrdU-positive cells of BMSCs group (88±23), (45±13) (P<0.05) in 7 day and 14 day, and were higher than the latter.Conclusion:①G-CSF of intraperitoneal injection and BMSCs of tail vein injection could improve motor function in MCAO rats. Motor function of MCAO rats of G-CSF were improved significantly higher than that of BMSCs.②cognitive function could be improved by G-CSF of intraperitoneal injection and BMSCs of tail vein injection in rats. The improvement of cognitive function of MCAO rats of G-CSF were significantly higher than those of BMSCs by the tail vein injection.③Intraperitoneal injection of G-CSF can mobilize bone marrow stem cells, including BMSCs migrated to the surrounding cerebral ischemia. G-CSF can promote BMSCs to migrate to the corpus callosum. The results suggest that G-CSF may be better than stem cell transplantation for treatment of cerebral ischemia by mobilized bone marrow stem cells to cerebral ischemia area for clinical treatment. |
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