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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(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。
Objective To investigate the therapeutic effect and protective mechanism of intraventricular transplation of human umbilical cord mesenchymal stem cells (hUC-MSCs) on hypoxic-ischemic brain damage (HIBD) in neonatal rats. Methods The umbilical cord of 1 normal full-term healthy infant born in obstetrics of our hospital was collected under aseptic conditions. The hUC-MSCs were cultured and stained with BrdU. The differentiation of cultured MSCs (N = 30), HIBD group (n = 36) and MSCs group (n = 32). The HIBD group and MSCs group HIBD model was established in neonatal rats. After successful 24 h injection, labeled hUC-MSCs were injected into the right ventricle of rats in MSCs group. The growth and development of rats were recorded within 3 weeks after transplantation. Longa score Neurobehavioral evaluation of hUC-MSCs were observed by immunofluorescence survival, migration, differentiation and differentiation. Results After transplantation, the weight of rats in transplantation group increased more than that in control group (P <0.05), and the Longa score in transplantation group was less than that in control group at 2 and 3 weeks after transplantation ). Within 3 weeks after transplantation, BrdU positive cells could be found in rat brain tissue sections, mainly distributed in the hippocampus and cerebral cortex. Within 3 weeks after transplantation, glial fibrillary acidic protein (GFAP) Or the overall signal intensity of neuron-specific enolase (NSE) is gradually increased. Conclusions When hUC-MSCs are transplanted into neonatal rats, the transplanted hUC-MSCs can migrate to the injured site and differentiate into neuron-like cells, which can promote the differentiation of endogenous neurons and reflect a certain degree of brain protection.