协和医学杂志

2011, v.2(01) 19-23

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移植骨髓间充质干细胞对大鼠脑梗死后神经元凋亡的影响
Effect of Transplantation of Bone Marrow-derived Mesenchymal Stem Cells on the Apoptosis of Neurons in Ischemic Rats

包新杰;魏俊吉;冯铭;卢珊;赵春华;王任直;
BAO Xin-jie1,WEI Jun-ji1,FENG Ming1,LU Shan2,ZHAO Chun-hua2,WANG Ren-zhi11Department of Neurosurgery,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730,China 2Center in Tissue Engineering,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730,China

摘要(Abstract):

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。
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.

关键词(KeyWords): 脑梗死;间充质干细胞;神经元;凋亡
cerebral ischemia;mesenchymal stem cells;neuron;apoptosis

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基金项目(Foundation): 国家863计划重点项目(2006AA02A115)

作者(Author): 包新杰;魏俊吉;冯铭;卢珊;赵春华;王任直;
BAO Xin-jie1,WEI Jun-ji1,FENG Ming1,LU Shan2,ZHAO Chun-hua2,WANG Ren-zhi11Department of Neurosurgery,Peking Union Medical College Hospital,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730,China 2Center in Tissue Engineering,Institute of Basic Medical Sciences,Chinese Academy of Medical Sciences & Peking Union Medical College,Beijing 100730,China

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