| ObjectivePreparation of rat MCAO (Middle cerebral artery occlusion, MCAO) model, BMSCs(Bone marrow mesenchymal stem cells, BMSCs) transplantation in treatment of ischemicstroke rats, neural function of rats are observed,and the changes of inflammatory factorsIL-l0, TGF-β1in serum and brain tissue are detected, study of BMSCs transplantation onrats during cerebral ischemia and reperfusion injury repair effect and the possiblemechanismMethodChoose weight about100g male SD rats and break its neck to death,then extractits long bone marrow cells, through the whole bone marrow culture method to cultivateBMSCs, select three to five generation of cell diluted with saline water and constantvolume made of2x106/ml cell suspension liquid for later use.Using the improved Longe bolt legal system for Middle cerebral artery occlusion(MCAO) model of SD rat. The36healthy adult male SD rats are randomly divided intothree groups, group A as normal control group (n=4), gave no treatment; group B as modelgroup (n=16), after the success of modeling to isodose physiological saline injection; groupC as cell transplantation group (n=16),after the success of modeling, BMSCstransplantation, ischemia2h, reperfusion after24hours,2×106MSCs through tail veintransplantation into the MCAO rats. In1,3,7,14days after transplantation to observeneurobehavioral score, by elisa kit and Western blot method to detect the change ofinflammation factor IL-l0、TGF-β1in the rats serum and brain tissue. Result1. Successfully developed the SD rat BMSCs, with optical microscope, the originalgeneration of BMSCs attached quantity is less, the majority are round or oval, afterpurification, amplification, adherent cells gradually increased, show the colony growth,long spindle shaped; With the increase of number of batches, to6~7generation, cell aginggradually, slowing growth, form gradually converge, is given priority to with flat cells, Thecell of body is full. and the immunofluorescence for detection of BMSCs surface specificmarker training: CD34(-), CD54(+).2. The score of nerve behavior: group A0+0.00，each time point of1days: group B3.50+0.27, group C3.21±0.57;3days: group B3.45+0.45, group C3.01±0.35;7days:group B3.21+0.25, group C2.72±0.34;14days: group B2.83+0.52, group C2.31±0.24.groups B and C score was significantly higher than group A (P <0.01).1days in group Band group C show no significant difference (P>0.05).3days,7days and14days in thegroup C is significantly lower than group B, the difference is statistically significant(P<0.05).3. Changes of serum IL-10of MCAO rats: group A10.11±0.55, at different timepoints:1days: group B35.23±1.27, group C55.51±2.51;3days: group B28.82±0.78,group C:46.55±0.91;7days: group B25.18±0.53, group C37.12±1.38;14days; groupB16.09±1.26, group C28.43±0.62. groups of B and C of serum IL-10level issignificantly higher in group A (P <0.01).1,3,7,14days group, the expression of IL-10down gradually, but the serum level of IL-10,C group is obviously higher than that ofgroup B (P <0.01), the difference is statistically significant.4. Changes of serum TGF-β1of MCAO rats: group A16.00±0.81, at different timepoints:1days: group B69.21±1.25, group C88.38±0.67;3days: group B52.32±1.24,group C:73.77±1.02;7days: group B42.71±0.54, group C54.62±1.72;14days; group B33.32±1.24, group C40.72±0.99.1,3,7,14days in group B and C, the serum TGF-β1levels are significantly higher than that in group A (P<0.01). In group C, the serum TGF-β 1levels is significantly higher than that in group B (P<0.01), the difference is statisticallysignificant. And with1,3,7,14days time, group of B,C of serum TGF-β1expression isgradually lower5. The brain tissue of rats with MCAO IL-10, TGF-β1expression: Group A is lowexpression.group of B C, the expression of rats brain IL-10、TGF-β1is significantlyhigher than that group A (P<0.01),1,3,7,14days to take group B and group C rat braintissue to detect IL-10, TGF-β1expression is visible, with the passage of time, theexpression of IL-10, TGF-β1gradually reduced, but group B of IL-10, TGF-β1lowerthan group C obviously (P<0.01), the difference was statistically significant.Conclusion1．Bone marrow mesenchymal stem cells can treating ischemic brain injury;2．The mechanism of bone marrow mesenchymal stem cells to repair the damagedischemic brain injury may be through the secretion of TGF-β1、 IL-10and otherAnti-inflammatory factors. |
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