人脐血间充质干细胞移植对新生大鼠缺氧缺血性脑损伤的神经保护  

Neuroprotective effect and mechanism of human umbilical cord blood mesenchymal stem cells on hypoxic-ischemic brain injury in neonatal rats

作　　者：周玉[1] 莫国梁[1] 赵婧[1] 方平[1] Zhou Yu;Mo Guoliang;Zhao Jing;Fang Ping(Department of Pediatrics, Affiliated Hospital of North Sichuan Medical College, Nanchong637000, Sichuan Province, China)

机构地区：[1]川北医学院附属医院儿科,四川省南充市637000

出　　处：《中国组织工程研究》2019年第13期2035-2041,共7页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(81300528);项目负责人:赵婧;四川省科技计划项目(2017JY0115);项目负责人:赵婧~~

摘　　要：背景:目前研究虽已证实脐血间充质干细胞移植可促进神经功能的恢复,但其具体作用机制尚不明确。目的:观察人脐血间充质干细胞移植对新生大鼠缺氧缺血性脑损伤的治疗作用,分析其可能的作用机制。方法:取10 d龄新生SD大鼠(由川北医学院实验动物中心提供),置于6 000 m海拔高原低压环境模拟仓生活和运动(运动方式为在舱内游泳槽内进行为期15 d的运动,60 min/d),建立高原缺血缺氧性脑损伤模型。造模成功24 h后,将60只大鼠随机分为移植组、模型组,移植组脑组织海马内注射DAPI标记的脐血间充质干细胞悬液,模型组以同样方法注射等量PBS;另取30只大鼠作为空白对照组,不建模不移植。细胞移植48 h后,TUNEL法检测神经细胞凋亡,Western blot法检测脑组织Caspase-3、p-Akt蛋白表达;细胞移植后21 d,Morris水迷宫实验测试大鼠学习记忆能力;细胞移植28 d后,ELISA法检测大鼠脑组织炎性因子表达,组织学观察脑组织病理变化。结果与结论:①TUNEL染色显示空白对照组仅见极少量凋亡细胞,模型组可见大量细胞凋亡,移植组细胞凋亡介于空白对照组与模型组之间,3组间细胞凋亡数量比较差异有显著性意义;②移植组脑组织Caspase-3蛋白表达明显低于模型组(P <0.05),p-Akt蛋白表达明显高于模型组(P <0.05);③Morris水迷宫实验测试结果显示,移植组大鼠逃避潜伏期与游泳距离均明显短于模型组(P <0.05);④与模型组比较,移植组肿瘤坏死因子α、白细胞介素1β、白细胞介素10质量浓度明显降低(P <0.05),白细胞介素8质量浓度明显升高(P <0.05);⑤模型组大鼠海马区神经元水肿,核染色质结构不清,有空泡形成,室管膜细胞重度水肿;移植组大鼠海马区神经元水肿程度轻于模型组,核染色质结构不清,未见空泡形成;⑥结果提示,人脐血间充质干细胞可通过调控炎性因子的表达、抑制神经细胞凋亡来改善新生大�BACKGROUND: Although it has been confirmed that human umbilical cord blood mesenchymal stem cells transplantation can promote the recovery of neural function, its mechanism is unclear.OBJECTIVE: To observe the therapeutic effect of human umbilical cord blood mesenchymal stem cells transplantation on hypoxic-ischemic brain injury in neonatal rats and to analyze its possible mechanism.METHODS: Sixty 10-day-old neonatal Sprague-Dawley rats(provided by the Experimental Animal Center of North Sichuan Medical College in China) were placed in a simulated hypobaric environment at 6 000-meter altitude to live and exercise. The animal model of ischemic-hypoxic brain injury at high altitude was established by exercising 60 min/d for 15 days in the swimming tank of the cabin. At 24 hours after modeling, 60 rats were randomly divided into transplantation group and model group. In the transplantation group, DAPI labeled human umbilical cord blood mesenchymal stem cells were injected into the rat hippocampus, while in the model group, the same volume of PBS was injected into the rat hippocampus. Another 30 rats were taken as blank control group, with no modeling and treatment. At 48 hours after cell transplantation, apoptosis of neurons was detected by TUNEL method, and western blot assay was used to detect the expression of Caspase-3 and p-Akt protein in brain tissue. At 21 days after cell transplantation, the learning and memory abilities of rats were measured by Morris water maze test. At 28 days after cell transplantation, the expression of inflammatory factors in rat brain tissue was detected by ELISA and pathological changes of the rat brain tissue were observed histologically.RESULTS AND CONCLUSION:(1) The results of TUNEL staining showed that only a small number of apoptotic cells were observed in the blank control group, and a large number of apoptotic cells were observed in the model group. The number of apoptotic cells in the transplantation group was higher than that in the blank control group but lower than that in

关 键 词：脐血间充质干细胞 细胞移植 神经保护 缺氧缺血性脑损伤 CASPASE-3蛋白 p-Akt 蛋白 细胞凋亡 国家自然科学基金 脐血干细胞移植 缺氧缺血 脑 半胱氨酸天冬氨酸蛋白酶3 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R392.4[医药卫生—临床医学]