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目的观察骨髓间充质干细胞(bone marrow stromal cells,BMSCs)经静脉注射移植对缺血/再灌注脑内神经细胞增殖和分化的影响。方法体外培养和扩增成年雄性大鼠BMSCs;以“四管阻断法”制作大鼠前脑缺血/再灌注模型;造模后3、5、7d通过尾静脉注射Hoechst33342标记的BMSCs(2×106/1ml/只),另设对照组于相同时间点通过尾静脉注射载体溶液。于造模后4周处死大鼠,取脑切片,HE染色观察大鼠海马缺血性损伤情况;BrdU/GFAP、BrdU/MAP2和BrdU/Hoechst33342免疫荧光双标染色,观察大鼠脑内神经细胞的增殖及分化情况。结果BMSCs移植组海马CA1区存活锥体细胞数显著多于载体溶液对照组(P<0.05)。BMSCs移植大鼠脑内海马结构BrdU阳性细胞数显著多于载体溶液对照组(P<0.05),BrdU/MAP-2双标阳性细胞占BrdU阳性细胞的比例显著高于载体溶液对照组(P<0.05)。结论BMSCs经静脉注射移植能够减轻缺血性脑损伤,促进宿主脑内自体神经细胞增殖,并提高其分化为神经元样细胞的比例。
Objective To observe the effects of bone marrow stromal cells (BMSCs) transplanted on the proliferation and differentiation of neural cells in the brain after ischemia / reperfusion. Methods BMSCs of adult male rats were cultured and expanded in vitro. The model of forebrain ischemia / reperfusion was established by “four tube occlusion method”. BMSCs labeled with Hoechst33342 (2 × 106 / 1ml / only), and another control group at the same time point through the tail vein injection of vehicle solution. The rats were sacrificed 4 weeks after the model was established. Brain slices were taken for HE staining to observe the hippocampal ischemic injury. BrdU / GFAP, BrdU / MAP2 and BrdU / Hoechst33342 immunofluorescence double staining were used to observe the changes of nerve cells Proliferation and differentiation. Results The number of surviving pyramidal cells in hippocampal CA1 area of BMSCs transplantation group was significantly more than that of vehicle control group (P <0.05). The number of BrdU positive cells in hippocampus of BMSCs transplantation group was significantly higher than that of vehicle control group (P <0.05), and the ratio of BrdU positive cells to BrdU positive cells in BrdU / MAP-2 positive group was significantly higher than that in vehicle control group (P < 0.05). Conclusion BMSCs transplanted intravenously can reduce ischemic brain injury, promote the proliferation of autologous nerve cells in the brain and increase the percentage of neurons that differentiate into neurons.