Xu Ming, Xu Hongzhi, Qin Zhiyong, Zhang Jie, Yang Xiaoyu, Xu Feng
Department of Anesthesiology, Fudan University, Shanghai, China.
Cell Biochem Biophys. 2014 Sep;70(1):443-7. doi: 10.1007/s12013-014-9937-0.
To compare the mRNA level of angiogenic factor vascular endothelial growth factor (VEGF), matrix metalloproteinases (MMP)-2, and MMP-9 in cultured human brain arteriovenous malformation (AVM) endothelial cells (ECs) and normal brain endothelial cells (BECs). Tissue explants both from deformed vessels of AVM and normal microvessel were put into culture for endothelial cells. After the monolayer adherent ECs reached confluence, they were tested with endothelial specific marker CD34 and von Willebrand factor (vWF) by immunochemical assay. mRNA levels of VEGF-A, MMP-2, and MMP-9 in AVM endothelial cells (AVMECs) and BECs were measured by PCR. Immunostaining confirmed that more than 95 % of the cultured cells were CD34 (Fig. 1b) and/or vWF positive. Expression levels of VEGF-A and MMP-2 mRNAs were significantly higher in AVMECs than in BECs. The MMP-9 level was also increased in AVMECs, but the difference was not statistically significant. Vascular tissue explants adherent method is a better approach for isolation and culture of AVMECs. Cultured AVMECs expressed higher angiogenic factors (VEGF, MMP-2) than the controlled BECs, implicating angiogenesis plays an important role in the pathogenesis of AVM.
比较培养的人脑动静脉畸形(AVM)内皮细胞(ECs)和正常脑内皮细胞(BECs)中血管生成因子血管内皮生长因子(VEGF)、基质金属蛋白酶(MMP)-2和MMP-9的mRNA水平。将来自AVM畸形血管和正常微血管的组织外植体进行培养以获得内皮细胞。单层贴壁ECs达到汇合后,通过免疫化学分析用内皮特异性标志物CD34和血管性血友病因子(vWF)进行检测。通过PCR测量AVM内皮细胞(AVMECs)和BECs中VEGF-A、MMP-2和MMP-9的mRNA水平。免疫染色证实超过95%的培养细胞为CD34阳性(图1b)和/或vWF阳性。AVMECs中VEGF-A和MMP-2 mRNA的表达水平显著高于BECs。AVMECs中MMP-9水平也有所升高,但差异无统计学意义。血管组织外植体贴壁法是分离和培养AVMECs的更好方法。培养的AVMECs比对照BECs表达更高的血管生成因子(VEGF、MMP-2),这表明血管生成在AVM的发病机制中起重要作用。