| Objectives This study through the animal experiment preliminary exploration of repeat bone marrow mesenchymal stem cells transplantation to treat cerebral infarction can improve its neural functional prognosis, reduce the cerebral infarction area of experimental animals, is a more effective cell therapy, as well as the possible mechanism of action, clinical application of cell therapy for treatment of cerebral infarction.Methods 1 The young SD rat bone marrow were cultured, using the whole bone marrow adherence screening method, separation, purification of BMSCs training. Take the third or 4 generation cells for transplantation, transplantation, 24 hours before the final concentration of 10umol/l labeled by Brd U.2 The establishment of rat model of cerebral infarction with modified Longa suture method, according to the m NSS selected 60 rats, were randomly divided into control group and single group, with 20 rats in each group, repeat group after modeling in 24 h, the control group were injected with 1ml saline group, single and repeat group were injected with 3×106 BMSCs for 7 days after modeling, control group and single group were injected with 1ml saline injection 3×106, repeat group BMSCs were injected through the tail vein.3 The results of immunohistochemical staining, the expression of VEGF levels lower than control group in a single group and reinsurance, statistically significant difference(P<0.05), single and repeat group significantly(P<0.05);Control group is lower than the single group of brain Olig- 2 and repeat group and a set of relative to the control group(P<0.01), single and repetitive control group(P<0.01), statistically significant difference, the control brain the Bcl- 2 expression level lower than that of single group and reinsurance, statistically significant difference(P<0.05), single and repeat group significantly(P< 0.05);Control group Caspase- 3 protein expression is higher than single and repeat groups, statistically significant difference(P<0.05).4 After modeling rats were sacrificed at 14 d, immunohistochemical staining was performed to observe the Brdu labeling rate, single staining observation of VEGF, Olig-2, Bcl-2, Caspase-3 protein, observe the neovascular endothelial cells, oligodendrocyte cell proliferation, cell apoptosis.6 Western blot:protein content of Bcl-2 and Caspase-3.Results 1 mNSS:3d after modeling, a single group and repeat group than in the control group m NSS, the difference was statistically significant(P<0.05), repeat group and single m NSS group, the difference was statistically significant(P<0.05), model 7d, single and repeat group than in the control group, m NSS low, the difference was statistically significant(P<0.01), model 14 d, single group, repeat m NSS group were significantly lower than the control group, the difference was statistically significant(P<0.01), single repeat group than in group m NSS, the difference was statistically significant(P<0.05).2 7 days after modeling, the single group and repeat group relative infarct volume was less than that of the control group, the difference was statistically significant(P<0.05), the difference between the single and repeat group; 14 days aftermodeling, the single group and the control group, the relative infarct volume comparison, the difference was statistically significant(P<0.05), repeat group and control group, the difference was statistically significant(P<0.01), repeat group compared with the single group, the difference was statistically significant(P<0.05).4 Western blot showed: the control group was lower than that of Bcl-2 protein in brain tissue of a single group and repeat group, the difference was statistically significant(P<0.01), single and repeat groups was statistically significant(P<0.05); the control group was higher than that of Caspase-3 protein in brain tissue of a single group and repeat group, the difference was statistically significant(P<0.01) a single group, and repeat groups was statistically significant(P<0.05), suggesting that BMSCs may promote cell proliferation, vascular stimulation of oligodendrocyte regeneration, reduce apoptosis in treatment of cerebral infarction.Conclusions 1 The whole bone marrow adherent method can rapidly proliferate, differentiate and purify BMSCs. 2 Allogeneic BMSCs has the therapeutic effect for cerebral infarction in rats, and the effect of repeated transplantation is better than that of single transplantation. 3 The forward treatment effect of BMSCs is better. 4 The model of permanent cerebral infarction was relatively simple, repeatable and less damage. 5 Brd U labeled BMSCs, simple operation, high labeling rate, the cell morphology, growth, etc. 6 BMSCs can migrate to cerebral ischemia region and no adverse reactions were found. 7 BMSCs may stimulate VEGF protein secretion, promote blood vessel regeneration, improve the blood circulation of brain tissue, Olig-2 protein secretion increased, stimulation of oligodendrocyte cell proliferation, enhanced axonal conduction ability, and up regulate the expression of Bcl-2 protein, down caspase-3 protein inhibits apoptosis and less infarction volume mechanism in the treatment of cerebral infarction. |
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