摘要
背景:骨髓间充质干细胞移植技术对脑损伤修复具有重要的意义,但其作用途径尚需讨论。目的:观察脑缺血模型大鼠移植骨髓间充质干细胞后,皮质及海马区Bcl-2和Bax的表达变化,分析骨髓间充质干细胞颅内的移植抑制神经元凋亡的作用机制。设计:随机对照实验。单位:青岛大学医学院附属医院脑血管病研究所。材料:选用雄性成年Wistar大鼠24只,体质量180～240g,由山东大学实验动物中心提供实验大鼠随机分为对照组、损伤组、损伤移植组,每组8只。溴脱氧核苷尿嘧啶(BrdU,Sigma公司);TUNEL试剂盒,Bcl-2,Bax抗体试剂盒购自北京中山生物工程公司。方法:实验于2005-04/2006-09在青岛大学医学院附属医院脑血管病研究所完成。损伤组和移植组大鼠采用颈外动脉线栓法建立脑缺血模型,移植组大鼠造模成功7d后在脑皮质和纹状体区注射浓度为2×10^(12)L^(-1)的原代体外培养的骨髓间充质干细胞悬液。实验过程中对动物的处置符合动物伦理学要求。主要观察指标:分别于造摸成功后7和14d进行TUNEL染色及免疫组织化学检测大脑皮质、海马区神经元凋亡和Bcl-2,Bax表达的变化。结果:①神经元凋亡情况:造模成功后7d,对照组来发现凋亡细胞,移植组凋亡细胞的数量少于损伤组,差异有非常显著性意义(P<0.01)。②Bcl-2和Bax表达:造模成功后14d,损伤组大鼠皮质和海马区Bcl-2阳性神经元的表达低于对照组和移植组,差异有非常显著性意义(P<0.01)。损伤组大鼠皮质和海马区Bax阳性神经元的表达高于对照组和移植组,差异有非常显著性意义(P<0 01)。结论:大鼠脑缺血后,骨髓间充质干细胞能够促进Bcl-2表达并抑制Bax表达,从而减少神经元凋亡。
BACKGROUND： The implantation technique of bone marrow mesenchymal stem cells （MSCs） is of important significance for repair of brain injury. However, its action pathway still needs to be investigated.
OBJECTIVE： To observe the changes of Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus of rats with cerebral ischemia in which MSCs were implanted, and to analyze the action mechanism of intracranial implantation of MSCs inhibiting neuronal apoptosis.
DESIGN： A randomized controlled experiment.
SETTING ： Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College.
MATERIALS： Twenty-four male adult Wistar rats, weighing 180 to 240 g, were provided by the Experimental Animal Center of Shandong University. The involved rats were randomized into 3 groups with 8 in each： control group, injury group and implantation group. 5-bromodeoxyuridine （BrdU, Sigma Company）, TUNEL kit, Bcl-2, Bax antibody kit were purchased from Beijing Zhongshan Bioengineering Company.
METHODS ： This study was carried out in the Institute for Cerebrovascular Disease, the Affiliated Hospital of Qingdao University Medical College between April 2005 and September 2006. Rats in the injury group and implantation group were developed into rat models of cerebral ischemia by suture of external carotid artery. Seven days later, the successful rat models in the implantation group were injected in the cerebral cortex and striatum with 2×10^12 L^-1 MSCs suspension primarily cultured in vitro. The processing of the experimental animals corresponded to the requests of Animal Ethics.
MAIN OUTCOME MEASURES ： Neuronal apoptosis and Bcl-2 and Bax expressions in the injured regions of cerebral cortex and hippocampus were detected by TUNEL staining and immunohistochemical method on the 7 and 14 days after successful modeling, separately.
RESULTS ： ①Neuronal apoptosis： On the 7^th day after successful modeling, apoptotic cells were not found in the con
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2007年第50期10205-10208,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
