| Aims: To observe the expression of mi RNA-21 and Bcl-2 in cerebral tissue at different times of I/R injury in rats transplanted with BMSCs, and to investigate the therapeutic mechanism of BMSCs during cerebral I/R injury as well as its relationship with mi R-21 and Bcl-2 upregulation.Method: Establishment of a rat middle cerebral artery ischemia(MCAO) and reperfusion model.96 health Sprague-Dawley(SD) rats are randomly divided into the following groups. They are respectively sham-operation group(24 ones), namely blank control group which accepts molding process but do not conduct artery embolism; model group(24 ones) which is not processed after successful molding; PBS transplantation group(24 ones) which is injected with PBS through caudal vein within 6 hours after the successful molding; BMSCs transplantation group(24 ones) in which BMSCs(cell concentration of 1×109/L) are transplanted through caudal vein within 6 hours after successful molding. HE staining is conducted; the expression level of micro RNA-21 is detected with RT-PCR method; the Bcl-2 protein content is explored with immunohistochemical method. Moreover, the expression of m NSS score, micro RNA-21, Bcl-2 protein contents of each group is analyzed; the relationship of each experimental group is analyzed..Results:1.SD rat m NSS score: blank group:0;According to 12 h, 1d, 3d, 7d at different time points, model group: 15.17±3.60 、 15.50±1.64 、 14.33±2.16、13.50±3.09 PBS group: 15.17±0.75、14.00±2.61、13.17±2.48、13.00±1.40;BMSCs group: 47.29±4.78、 98.69±7.50、78.72±5.79、61.35±5.90. At 12 h, 1d three groups of rats m NSS scores were not statistically different(P>0.05), then each group scores gradually reduced,There was significant difference between BMSCs group before and after(P<0.05); at the same time 3d, 12 d, BMSCs group decreased more obviously, compared with PBS group, model group, the two groups had significant difference(P<0.01); there is no significant difference PBS Group compared with the model group(P>0.05).. 2. Under light microscope the hippocampus of the rat HE staining showed:blank group: There was no obvious pathological change,Neural cell number, neat, uniform, morphological integrity, the patina bright red, the nucleus round or oval. model group, PBS transplantation group:Massive necrosis of nerve cells,Cell swelling, loss, Obvious necrosis, the remaining cell shrinkage,the patina pale staining, the nucleus pyknosis. BMSCs group : Necrosis area to reduce.. 3.Bcl-2 immunohistochemical results:According to the time sequence of 12 h, 1d, 3d, 7d at different time points, to observe the brain tissue in the average number of each individual field of vision, the positive expression of Bcl-2 in blank group: 5.69±0.56, 5.16±0.78, 5.43±0.67, 5.17 ±0.66; model group: 24.51±3.65, 53.80±5.71, 45.14±3.13, 34.37±3.16; PBS group: 26.3± 2.56,57.62±6.14,43.56±2.30,34.38±2.63;BMSCsgroup:47.29±4.78,98.69±7.50, 78.72 ±5.79, 61.35±5.90. Through statistical testing found:In the 12 h to 1d range, Bcl-2 with the prolongation of the time each positive cell number were increased after 1d, Bcl-2 positive number began to decrease(P<0.05). The same time points in model group compared with the blank group, the number of Bcl-2 positive cells was more obvious(P<0.05); BMSCs group compared with model group, the number of Bcl-2 positive cells was more obvious (P<0.05); BMSCs group and PBS group compared to the number of Bcl-2 positive cells was more obvious(P<0.05); PBS group compared with the model group, the number of Bcl-2 positive cells had no statistical significance(P>0.05). 4.Quantitative Real-time PCR for detection of mi R-21:According to 12 h, 1d, 3d, 7d at different time points, blank group:1±0.03; model group: 1.35±0.08, 2.19±0.04, 1.80±0.10,1.37±0.10; PBS group: 1.39±0.12, 2.19±0.11, 1.74±0.12,1.31±0.10;BMSCs group:1.64±0.10,3.49±0.17,2.69±0.07, 1.96± 0.18. In the 12 h to 1d range of time, prolonged expression of mi R-21 with time in each group were increased, mi R-21 expression was decreased in each group after 1d(P<0.05). Each time point of the model group compared with blank group expression of mi R-21 was significantly higher(P<0.05); PBS group higher expression of mi R-21 compared with the model group but without statistical significance(p>0.05).BMSCs transplantation group and the other three groups, the expression of mi R-21 was significantly higher than the other three groups(P<0.01).Conclusion:1. Bone marrow mesenchymal stem cells attenuate the damage of neurological functions induced by cerebral ischemia reperfusion. 2. The mechanism underlying the treatment of bone marrow mesenchymal stem cells against cerebral ischemia-reperfusion injury may be involved in upregulations of mi R-2 and Bcl-2. |
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