Institute of Medical Biotechnology, Department of Human Anatomy, Histology and Embryology, School of Biology and Basic Medical Sciences, Soochow University, and Children's Hospital Affiliated to Suzhou University, Jiangsu Province Key Laboratory of Stem Cells, Suzhou 215007, PR China.
Mol Med Rep. 2012 Sep;6(3):625-30. doi: 10.3892/mmr.2012.968. Epub 2012 Jun 26.
Cerebrovascular injury is one of the three major causes of death and is the leading cause of adult disability. Despite the increasing progress in emergency treatment and early rehabilitation in patients with cerebrovascular injury, treatment options for neurological dysfunction that presents at a later stage are lacking. This study examined the potential of human amniotic mesenchymal stem cell (hAMSC) transplantation in the repair of neurological deficits in an experimental focal cerebral ischemia model. Following the isolation of hAMSCs, growth characteristics and surface antigen expression were observed. Butylated hydroxyanisole (BHA) was used to induce the cultured cells into neuron-like cells, which were identified by immunocytochemistry. The suture model was used to induce focal cerebral ischemia in rats, which were subsequently randomly divided into experimental and control groups for treatment with BrdU-labeled hAMSCs or PBS, respectively. Neurological deficits were assessed following transplantation using the neurological severity score, beam balance test and elevated body swing test. Eight weeks later, rat brain tissue was analyzed with H&E staining and BrdU immunohistochemistry, and the survival and spatial distribution of the transplanted hAMSCs were determined. The hAMSCs proliferated in vitro, and it was found that neuron-specific enolase (NSE) was expressed in neurons, whereas glial fibrillary acidic protein (GFAP) was expressed in astrocytes. The focal ischemia model caused varying degrees of left limb hemiplegia accompanied by right sided Horner's Syndrome. When examined 1, 3, 6 and 8 weeks later, significant recovery in neurological behavior was detected in the rats treated with hAMSC transplantation compared with the control (P<0.01). BrdU-labeled hAMSCs were concentrated near the graft site and surrounding areas, in certain cases migrating towards the ischemic lesion. Local gliosis and lymphocytic infiltration were not detected. hAMSCs exhibit great potential for proliferation and are induced to differentiate into NSE-expressing neuron-like cells following treatment with BHA. Moreover, hAMSC transplantation may improve neurological symptoms following focal cerebral ischemia.
脑血管损伤是死亡的三大原因之一，也是成年人残疾的主要原因。尽管在脑血管损伤患者的紧急治疗和早期康复方面取得了越来越多的进展，但对于后期出现的神经功能障碍，缺乏有效的治疗选择。本研究探讨了人羊膜间充质干细胞（hAMSC）移植在实验性局灶性脑缺血模型中修复神经功能缺损的潜力。在分离 hAMSCs 后，观察其生长特性和表面抗原表达。用丁基羟基茴香醚（BHA）诱导培养细胞分化为神经元样细胞，并用免疫细胞化学法进行鉴定。采用缝线法诱导大鼠局灶性脑缺血，随后随机分为实验组和对照组，分别用 BrdU 标记的 hAMSCs 或 PBS 治疗。移植后通过神经功能严重程度评分、平衡木试验和抬高身体摆动试验评估神经功能缺损。8 周后，用 H&E 染色和 BrdU 免疫组织化学法分析大鼠脑组织，并确定移植的 hAMSCs 的存活和空间分布。hAMSCs 在体外增殖，发现神经元特异性烯醇化酶（NSE）在神经元中表达，而胶质纤维酸性蛋白（GFAP）在星形胶质细胞中表达。局灶性缺血模型导致左侧肢体偏瘫伴右侧霍纳氏综合征，程度不一。1、3、6 和 8 周后检测发现，hAMSC 移植治疗组大鼠的神经行为有明显恢复，与对照组相比差异有统计学意义（P<0.01）。BrdU 标记的 hAMSCs 集中在移植物部位及其周围，在某些情况下向缺血病变部位迁移。未检测到局部神经胶质增生和淋巴细胞浸润。hAMSC 具有强大的增殖潜力，经 BHA 处理后可诱导分化为表达 NSE 的神经元样细胞。此外，hAMSC 移植可能改善局灶性脑缺血后的神经症状。
