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范 雪, 白小红, 陈 娟, 等.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.四川大学学报(医学版),2017,48(2):179-185
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
Protective Effects of Intraventricular Transplanted Human Umbilical Cord-derived Mesenchymal Stem Cells on Hypoxic Ischemic Brain Damages in Rats
  
中文关键词:  【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
英文关键词:【Key words】 Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) Hypoxia ischemic brain damage (HIBD) Cellular transplantation therapy Brain protection effect
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中文摘要:
      【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3~4 cm,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10 d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24 h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(P <0.05);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(P <0.05);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。
英文摘要:
      【Abstract】 Objective To determine the therapeutic effect and protective mechanism of human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) on newborn rats with hypoxia ischemic brain damage (HIBD). Methods Umbilical cord (3-4 cm) was collected from a healthy male infant for preparation of hUC-MSCs using explants technique. The hUC-MSCs were cultured and labeled with BrdU. The differentiation function of MSCs was identified. Healthy SPF grade neonatal SD rats were randomly divided into sham (n =30), HIBD (n =36) and hUC-MSCs treated HIBD (n =32) groups. BrdU-labeled hUC-MSCs were injected into the right ventricle of the rats in the hUC-MSCs treatment group 24 h after successful induction of HIBD. The growth and development of the rats were recorded. The neurological behavior of the rats were evaluated with Longa score method 3 weeks after hUC-MSCs transplantation. The survival, migration, differentiation and pro-differentiation of the transplanted hUC-MSCs were measured using immunological fluorescence method. Results Rats in the hUC-MSCs treatment group had significant higher weight gain and lower Longa scores (at the second and the third week post transplantation) than those in the HIBD group (P <0.05). BrdU positive cells were found in brain tissues 3 weeks after transplantation, and they were mainly distributed in the damaged hippocampus and cerebral cortex. Three weeks after transplantation, the total signal strength of glial fibrillary acidic protein (GFAP) or neuron-specific enclase (NSE) gradually increased. Conclusion Transplanted hUC-MSCs can migrate brain damages through differentiating into neuron-like cells and promoting endogenous neurological differentiations.
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