脑脉通联合骨髓干细胞动员对脑缺血大鼠神经细胞凋亡及Bax、Bcl-2表达的影响

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目的观察脑脉通联合骨髓干细胞(BMSCs)动员对脑缺血大鼠神经细胞凋亡的作用,并从对凋亡相关基因Bax、Bcl-2表达及Bax/Bcl-2变化的影响探讨其可能的作用机制。方法大鼠随机分组。线栓法制备局灶性脑缺血(MCAO)动物模型。动员组和联合组大鼠分别于术前3d和术后2d皮下注射人重组粒细胞集落刺激因子(rG-CSF);脑脉通灌胃用药。术后2、3、7、14d取材,测定外周血和脑组织CD34+细胞;观察脑组织病理改变;检测神经细胞凋亡;测定脑组织Bax、Bcl-2表达,计算Bax/Bcl-2比值。结果模型组大鼠外周血和脑组织CD34+细胞增加,分别于3d和7d达到峰值;各治疗组尤其联合组脑组织CD34+增强明显。缺血后7d大鼠脑组织损伤明显,动员组14d和各联合组的损伤减轻。各模型组神经细胞凋亡增多、Bax表达增加、2d和3d的Bax/Bcl-2增高。各联合组神经细胞凋亡减轻、Bax表达减弱,7d及14d的Bcl-2表达增强,各时间点的Bax/Bcl-2降低。联合组较动员组2d和3d的凋亡细胞、各时间点的Bax表达减轻和Bax/Bcl-2降低,7d及14d的Bcl-2表达增强。结论脑缺血可引起BMSCs进入外周血及向脑组织归巢,达到峰值的时间存在差异,脑脉通可使动员后的骨髓干细胞增多;脑缺血引起的神经细胞凋亡与Bax表达上调相关,脑脉通可使BMSCs动员抑凋亡的作用增强并延长,其作用可能与早期(2d、3d)下调促凋亡基因Bax和后期(7d、14d)上调Bcl-2表达有关。 Objective To observe the effect of Naomaitong combined with bone marrow stem cells (BMSCs) mobilization on neuronal apoptosis in rats with cerebral ischemia, and to explore its possible effects on the expression of apoptosis-related genes Bax, Bcl-2 and Bax/Bcl-2. The mechanism of action. Methods Rats were randomly divided into groups. Preparation of Focal Cerebral Ischemia (MCAO) Animal Model by Suture. Rats in the mobilization group and the combination group were injected subcutaneously with human recombinant granulocyte colony-stimulating factor (rG-CSF) 3 days before surgery and 2 days after surgery respectively; At the 2nd, 3rd, 7th, and 14th days after operation, CD34+ cells were detected in peripheral blood and brain; the pathological changes of brain tissue were observed; the apoptosis of nerve cells was detected; the expression of Bax and Bcl-2 in brain tissue was measured and the ratio of Bax/Bcl-2 was calculated. Results The CD34+ cells in the peripheral blood and brain tissue of the model group increased, peaked at 3d and 7d respectively. The CD34+ in the brain tissue of the treatment group, especially the combined group, increased significantly. At 7 days after ischemia, the brain tissue of rats was significantly damaged, and the injury in the mobilized group was reduced on the 14th day and the combined groups. In each model group, the apoptosis of neurons increased, the expression of Bax increased, and the Bax/Bcl-2 levels increased at 2d and 3d. The neuronal apoptosis and Bax expression were weakened in each combination group. The expression of Bcl-2 was increased at 7d and 14d, and Bax/Bcl-2 was decreased at each time point. Compared with the mobilized group, apoptotic cells at the 2nd and 3rd day in the combination group decreased Bax expression and Bax/Bcl-2 levels at each time point, and enhanced Bcl-2 expression at 7d and 14d. Conclusion Cerebral ischemia can cause BMSCs to enter peripheral blood and return to brain tissue, and there is a difference in the peak time. Naomaitong can increase the number of mobilized bone marrow stem cells; the neuronal apoptosis induced by cerebral ischemia is related to the up-regulation of Bax expression. Naomaitong can enhance and prolong the mobilization and apoptosis of BMSCs, which may be related to the down-regulation of pro-apoptosis gene Bax in early stage (2d, 3d) and up-regulation of Bcl-2 expression in late stage (7d, 14d).
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