Choi Seung Ah, Chong Sangjoon, Kwak Pil Ae, Moon Youn Joo, Jangra Anshika, Phi Ji Hoon, Lee Ji Yeoun, Park Sung-Hye, Kim Seung-Ki
1Division of Pediatric Neurosurgery, Pediatric Clinical Neuroscience Center, Seoul National University Children's Hospital.
2Department of Neurosurgery, Seoul National University Hospital, Seoul National University College of Medicine.
J Neurosurg Pediatr. 2019 Feb 1;23(2):204-213. doi: 10.3171/2018.8.PEDS1883. Epub 2018 Nov 9.
OBJECTIVEEndothelial colony-forming cells (ECFCs) isolated from pediatric patients with moyamoya disease (MMD) have demonstrated decreased numbers and defective functioning in in vitro experiments. However, the function of ECFCs has not been evaluated using in vivo animal models. In this study, the authors compared normal and MMD ECFCs using a chronic cerebral hypoperfusion (CCH) rat model.METHODSA CCH rat model was made via ligation of the bilateral common carotid arteries (2-vessel occlusion [2-VO]). The rats were divided into three experimental groups: vehicle-treated (n = 8), normal ECFC-treated (n = 8), and MMD ECFC-treated (n = 8). ECFCs were injected into the cisterna magna. A laser Doppler flowmeter was used to evaluate cerebral blood flow, and a radial arm maze test was used to examine cognitive function. Neuropathological examinations of the hippocampus and agranular cortex were performed using hematoxylin and eosin and Luxol fast blue staining in addition to immunofluorescence with CD31, von Willebrand factor, NeuN, myelin basic protein, glial fibrillary acidic protein, and cleaved caspase-3 antibodies.RESULTSThe normal ECFC-treated group exhibited improvement in the restoration of cerebral perfusion and in behavior compared with the vehicle-treated and MMD ECFC-treated groups at the 12-week follow-up after the 2-VO surgery. The normal ECFC-treated group showed a greater amount of neovasculogenesis and neurogenesis, with less apoptosis, than the other groups.CONCLUSIONSThese results support the impaired functional recovery of MMD ECFCs compared with normal ECFCs in a CCH rat model. This in vivo study suggests the functional role of ECFCs in the pathogenesis of MMD.
从烟雾病（MMD）患儿中分离出的内皮祖细胞（ECFCs）在体外实验中已显示数量减少且功能缺陷。然而，尚未使用体内动物模型评估ECFCs的功能。在本研究中，作者使用慢性脑灌注不足（CCH）大鼠模型比较了正常和MMD的ECFCs。
通过结扎双侧颈总动脉（双血管闭塞[2-VO]）建立CCH大鼠模型。将大鼠分为三个实验组：溶剂处理组（n = 8）、正常ECFC处理组（n = 8）和MMD ECFC处理组（n = 8）。将ECFCs注入小脑延髓池。使用激光多普勒血流仪评估脑血流量，并使用放射状臂迷宫试验检查认知功能。除了用CD31、血管性血友病因子、NeuN、髓鞘碱性蛋白、胶质纤维酸性蛋白和裂解的半胱天冬酶-3抗体进行免疫荧光外，还使用苏木精和伊红以及Luxol固蓝染色对海马体和无颗粒皮质进行神经病理学检查。
在2-VO手术后12周的随访中，与溶剂处理组和MMD ECFC处理组相比，正常ECFC处理组在脑灌注恢复和行为方面表现出改善。与其他组相比，正常ECFC处理组显示出更多的新生血管形成和神经发生，且细胞凋亡更少。
这些结果支持在CCH大鼠模型中，与正常ECFCs相比，MMD ECFCs的功能恢复受损。这项体内研究表明ECFCs在MMD发病机制中的功能作用。
