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Effect of Transplantation of Bone Marrow-derived Mesenchymal Stem Cells on the Apoptosis of Neurons in Ischemic Rats
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摘要:
目的 探讨移植骨髓间充质干细胞(bone marrow-derived mesenchymal stemcells, BMSCs)治疗大鼠脑梗死及其减轻半暗带神经元的凋亡作用。 方法 采集成人志愿者骨髓, 分离培养BMSCs; 线栓法建立大鼠大脑中动脉栓塞(middle cerebral artery occlusion, MCAO)2h再灌注脑梗死模型; 建模后第3天分别立体定向脑内移植5×105BMSCs和等量生理盐水, 改良神经功能损害评分(modified neurological severity scores, mNSS)观察神经功能恢复; 建模后第14天TUNEL法检测梗死周边神经细胞凋亡; 建模后第28天免疫荧光染色检测梗死周边神经元的存活; 采用依红-苏木素染色评估梗死体积。 结果 治疗组和对照组比较, 梗死周边TUNEL阳性细胞显著减少(P=0.017), 而神经元核抗原(neuronal nuclei antigen, NeuN)阳性细胞显著增多(P=0.015);同时, BMSCs可显著降低MCAO大鼠的相对梗死体积(P=0.041);mNSS评分显示, 从MCAO后第14天开始, 治疗组评分均显著低于对照组(P < 0.05)。 结论 脑梗死急性期移植BMSCs可显著减少半暗带神经元凋亡, 缩小梗死范围, 有效促进神经功能的恢复。 Abstract:Objective To investigate the apoptosis of neurons in the penumbra of cerebral ischemic rats transplanted with bone marrow derived-mesenchymal stem cells(BMSCs). Methods Bone marrow of a healthy adult volunteer was collected, and BMSCs were cultivated and harvested.Adult rats subjected to 2 h middle cerebral artery occlusion(MCAO)were randomly assigned to the BMSCs-treated group(n=12)and the control group(n=12), in which rats were transplanted with 5×105 BMSCs or saline, respectively, into the ipsilateral brain parenchyma on the 3rd post-MCAO day.The modified neurological severity scores(mNSS)was used to measure functional recovery.On the 14th post-MCAO day, apoptotic cells in the penumbra were detected by TUNEL analysis.On the 28th post-MCAO day, neurons were stained by immunoflu-orescence, and hematoxylin-eosin staining was used to evaluate the relative ischemic volume. Results TUNEL-positive cells were significantly reduced(P=0.017), and in contrast, neuronal nuclei antigen(NeuN)-positive cells were significantly increased(P=0.015)in the penumbra in BMSCs-treated group compared with control group.The relative ischemic volume was decreased in the BMSCs-treated group(28.3±4.5)% compared with the control group(36.8±5.6)%(P=0.041).The mNSS was significantly decreased in BMSCs-treated group at each time point starting on the 14th post-MCAO day compared with control group(P < 0.05). Conclusion BMSCs treatment can decrease ischemic volume and improve functional recovery after cerebral ischemia, which may, at least partially, attribute to the reduced apoptosis of neurons in the penumbra. -
Key words:
- cerebral ischemia /
- mesenchymal stem cells /
- neuron /
- apoptosis
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图 2 BMSCs治疗组和对照组梗死周边TUNEL阳性细胞比较
红色：TUNEL阳性细胞；蓝色：TUNEL细胞核；粉色：TUNEL阳性细胞与细胞核重叠后；A.对照组；B.治疗组；C.治疗组与对照组梗死周边TUNEL阳性细胞数量比较；与对照组比较，*P=0.017；图内标尺为75μm
图 3 BMSCs治疗组与对照组梗死周边神经元比较
红色：NeuN阳性细胞神经元；A.对照组；B.治疗组；C.治疗组和对照组梗死周边神经元数量比较；与对照组比较，*P=0.015；图内标尺为75μm
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