Department of Neurosurgery, Seoul National University College of Medicine, Seoul National University Hospital, Seoul;
J Neurosurg. 2014 Feb;120(2):415-25. doi: 10.3171/2013.9.JNS131000. Epub 2013 Oct 25.
Moyamoya disease (MMD) is a cerebrovascular occlusive disease affecting bilateral internal carotid termini. Smooth-muscle cells are one of the major cell types involved in this disease process. The characteristics of circulating smooth-muscle progenitor cells (SPCs) in MMD are poorly understood. The authors purified SPCs from the peripheral blood of patients with MMD and sought to identify differentially expressed genes (DEGs) in SPCs from these patients.
The authors cultured and isolated SPCs from the peripheral blood of patients with MMD (n = 25) and healthy control volunteers (n = 22). After confirmation of the cellular phenotype, RNA was extracted from the cells and DEGs were identified using a commercially available gene chip. Real-time quantitative reverse transcription polymerase chain reaction was performed to confirm the putative pathogenetic DEGs.
The SPC-type outgrowth cells in patients with MMD invariably showed a hill-and-valley appearance under microscopic examination, and demonstrated high α-smooth muscle actin, myosin heavy chain, and calponin expression (96.5% ± 2.1%, 42.8% ± 18.6%, and 87.1% ± 8.2%, respectively), and minimal CD31 expression (less than 1%) on fluorescence-activated cell sorter analysis. The SPCs in the MMD group tended to make more irregularly arranged and thickened tubules on the tube formation assay. In the SPCs from patients with MMD, 286 genes (124 upregulated and 162 downregulated) were differentially expressed; they were related to cell adhesion, cell migration, immune response, and vascular development.
With adequate culture conditions, SPCs could be established from the peripheral blood of patients with MMD. These cells showed specific DEGs compared with healthy control volunteers. This study provides a novel experimental cell model for further research of MMD.
烟雾病（MMD）是一种影响双侧颈内动脉终末的脑血管闭塞性疾病。平滑肌细胞是参与该疾病过程的主要细胞类型之一。MMD 患者循环中的平滑肌祖细胞（SPC）的特征尚未完全了解。作者从 MMD 患者的外周血中纯化 SPC，并试图鉴定这些患者的 SPC 中差异表达的基因（DEGs）。
作者从 MMD 患者（n=25）和健康对照志愿者（n=22）的外周血中培养和分离 SPC。在确认细胞表型后，从细胞中提取 RNA，并使用商业基因芯片鉴定 DEGs。实时定量逆转录聚合酶链反应（PCR）用于确认潜在的致病 DEGs。
MMD 患者的 SPC 型细胞外生长细胞在显微镜下始终呈现出丘陵和山谷的外观，并且表现出高水平的α-平滑肌肌动蛋白、肌球蛋白重链和钙调蛋白表达（分别为 96.5%±2.1%、42.8%±18.6%和 87.1%±8.2%），荧光激活细胞分选分析显示 CD31 表达水平极低（低于 1%）。MMD 组的 SPC 在管状形成测定中倾向于形成更不规则排列和增厚的小管。在 MMD 患者的 SPC 中，有 286 个基因（124 个上调和 162 个下调）差异表达；它们与细胞黏附、细胞迁移、免疫反应和血管发育有关。
在适当的培养条件下，可从 MMD 患者的外周血中建立 SPC。与健康对照志愿者相比，这些细胞显示出特定的 DEGs。本研究为进一步研究 MMD 提供了一种新的实验细胞模型。
