脑室内移植hUC-MSCs对缺氧缺血性脑损伤新生大鼠的保护作用

发布时间：2018-02-26 09:26

本文关键词： 人脐带间充质干细胞缺氧缺血性脑损伤细胞移植治疗脑保护作用　出处：《四川大学学报(医学版)》2017年02期 　论文类型：期刊论文

【摘要】：目的探讨脑室内移植人脐带间充质干细胞(hUC-MSCs)对新生大鼠缺氧缺血性脑损伤(HIBD)的治疗效果及保护性机制。方法无菌条件下采集在我院产科出生的1例正常足月健康男婴的脐带3~4cm,运用组织块贴壁法培养hUC-MSCs,使用BrdU标记细胞,并对培养出的MSCs的分化功能进行鉴定;取98只健康SPF级10d龄SD大鼠,随机分为假手术组(n=30)、HIBD组(n=36)和MSCs组(n=32),其中HIBD组和MSCs组建立新生大鼠HIBD模型,建模成功24h后将标记的hUC-MSCs注射入MSCs组大鼠右侧脑室,于移植后3周内,记录大鼠生长发育情况,并用Longa评分法对大鼠的神经行为学进行评价,用免疫荧光法观察移植hUC-MSCs的存活、迁移、分化及促分化情况。结果移植后,移植组大鼠体质量增加大于对照组,差异有统计学意义(P0.05);移植后2、3周,移植组Longa评分小于对照组,差异有统计学意义(P0.05);移植后3周内可在大鼠脑组织切片中发现BrdU阳性细胞,主要分布于损伤侧海马区域及大脑皮质;移植后3周内,大鼠脑组织中胶质纤维酸性蛋白(GFAP)或神经元特异性烯醇化酶(NSE)的总体信号强度逐渐增强。结论 hUC-MSCs移植治疗新生大鼠HIBD时,移植的hUC-MSCs可迁移至受损部位并分化为神经样细胞,可促进内源性神经分化,体现了一定程度的脑保护作用。
[Abstract]:Objective to investigate the therapeutic effect and protective mechanism of intracerebroventricular transplantation of human umbilical cord mesenchymal stem cells (hUC-MSCs) on hypoxic-ischemic brain injury (HIBD) in neonatal rats. The umbilical cord of the male infant was 3cm. The HUC-MSC cells were cultured by tissue mass adherent method. The cells were labeled with BrdU. 98 healthy SD rats of SPF grade 10 days old were randomly divided into sham-operation group (n = 30) and MSCs group (n = 32). The HIBD model of neonatal rats was established in HIBD group and MSCs group. After 24 hours of modeling, the labeled hUC-MSCs was injected into the right ventricle of the rats in the MSCs group. The growth and development of the rats were recorded within 3 weeks after transplantation. The neurological behavior of the rats was evaluated by Longa score, and the survival of the transplanted hUC-MSCs was observed by immunofluorescence. Results after transplantation, the body mass of rats in the transplantation group was significantly higher than that in the control group (P 0.05), and the Longa score in the transplantation group was lower than that in the control group 2 weeks after transplantation. The difference was statistically significant (P 0.05), BrdU positive cells could be found in the brain sections of the rats within 3 weeks after transplantation, mainly distributed in the injured hippocampus and cerebral cortex, and within 3 weeks after transplantation, the positive cells were found in the hippocampus and cerebral cortex of the injured side. The overall signal intensity of glial fibrillary acidic protein (GFAP) or neuron-specific enolase (NSE) in rat brain increased gradually. Conclusion the transplanted hUC-MSCs can migrate to the damaged site and differentiate into neuron-like cells after hUC-MSCs transplantation in neonatal rats with HIBD. Can promote endogenous neural differentiation, reflecting a certain degree of brain protection.
【作者单位】：四川大学华西第二医院儿科;出生缺陷与相关妇儿疾病教育部重点实验室;四川省干细胞库/四川新生命干细胞科技股份有限公司;
【基金】：国家自然科学基金(No.81330016,No.81630038) 教育部长江学者和创新团队基金项目(No.IRT0935)资助
【分类号】：R742

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