腺病毒介导脑源性神经营养因子与脑出血模型大鼠内源性神经干细胞的分化  

Adenovirus-mediated brain-derived neurotrophic factor and endogenous neural stem cell differentiation in a rat model of cerebral hemorrhage

作　　者：谢强[1] 王飞[1] 周国平[1] 张惠[2] 马进显[1] 

机构地区：[1]南阳市中心医院神经外科二病区,河南省南阳市473000 [2]南阳市中心医院耳鼻喉二病区,河南省南阳市473000

出　　处：《中国组织工程研究》2016年第27期4029-4035,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:以往的研究表明,神经营养因子具有多种功能,可对损伤后的神经元存活予以有效维持。目的:利用腺病毒对脑源性神经营养因子进行介导,观察其对大鼠脑出血后内源性神经干细胞分化的影响。方法:取90只SD大鼠,制备脑出血模型,脑出血后12 h开始腹腔注射5-溴脱氧尿嘧啶核苷,2次/d,连续注射10 d;造模成功后随机分为3组,每组30只,分别在脑部立体定位注射腺病毒载体、腺病毒介导脑源性神经营养因子、生理盐水,注射后1 d、3 d、1周、2周、3周、4周,评估神经功能缺失评分,检测脑出血后血肿周围区生长相关蛋白吸光度值;注射后4周,免疫双标检测5-溴脱氧尿嘧啶核苷/神经元、5-溴脱氧尿嘧啶核苷/神经胶质纤维酸性蛋白的表达。结果与结论:(1)随着时间的推移,3组神经功能缺失评分不断减小,腺病毒介导脑源性神经营养因子组注射1-4周的神经功能缺失评分低于腺病毒载体组、生理盐水组(P<0.05);(2)随着时间的推移,3组血肿周围区生长相关蛋白平均吸光度值先升高后降低,腺病毒介导脑源性神经营养因子组注射3 d-4周的血肿周围区生长相关蛋白平均吸光度值均高于腺病毒载体组、生理盐水组(P<0.05);(3)腺病毒介脑源性神经营养因子组5-溴脱氧尿嘧啶核苷/神经元、5-溴脱氧尿嘧啶核苷/神经胶质纤维酸性蛋白双阳性率显著高于腺病毒载体组、生理盐水组(P<0.05);(4)结果表明,利用腺病毒对脑源性神经营养因子进行介导并对脑出血大鼠进行干预,可有效促进内源性神经干细胞的分化,促进动物神经功能的恢复。BACKGROUND： Previous studies showed that neurotrophic factor has a variety of functions, which can effectively maintain the survival of neurons after injury. OBJECTIVE： To observe the effect of adenovirus-mediated brain-derived neurotrophic factor on the differentiation of endogenous neural stem cells after intracerebral hemorrhage in rats. METHODS： A total of 90 Sprague-Dawley rat models of cerebral hemorrhage were made. At 12 hours after cerebral hemorrhage, 5-bromodeoxyuridine（BrdU） was intraperitoneally injected, twice a day, for 10 consecutive days. After model establishment, rats were randomly divided into three groups, 30 rats in each group, and were respectively subjected to brain stereotaxic injection of adenovirus vector, adenovirus-mediated brain-derived neurotrophic factor and physiological saline. At 1 day, 3 days, 1 week, 2 weeks, 3 weeks, and 4 weeks, neurological deficit score was evaluated. Absorbance value of growth associated protein around the area of hematoma after intracerebral hemorrhage was measured. At 4 weeks after injection, double immunostaining was used to detect the expression of BrdU/NeuN and BrdU/glial fibrillary acidic protein（GFAP）. RESULTS AND CONCLUSION：（1） With the passage of time, nerve function defect score decreased in the three groups. At 1-4 weeks after injection, nerve function deficit scores were lower in the adenovirus-mediated brain-derived neurotrophic factor group than that in the adenovirus vector group and saline group（P〈0.05）.（2） With the passage of time, the average absorbance of three groups in the peri-hematoma region first increased and then decreased. The absorbance value was higher in the adenovirus-mediated brain-derived neurotrophic factor group than in the adenovirus vector group and saline group at 3 days-4 weeks（P〈0.05）.（3） BrdU/NeuN and BrdU/GFAP rates were significantly higher in the adenovirus-mediated brain-derived neurotrophic factor group than that of adenovirus vector group and saline group（P〈0.05）.�

关 键 词：脑出血 脑源性神经营养因子 神经干细胞 组织工程 实验动物 神经损伤与修复动物模型 腺病毒 病毒介导 内源性神经干细胞 

分 类 号：R318[医药卫生—生物医学工程]