论文部分内容阅读
目的研究人肾上腺微血管内皮细胞的表型和功能特性。方法用亚细胞克隆法分离和纯化人肾上腺微血管内皮细胞;流式细胞术检测内皮细胞标志血管性血友病因子(vWF)和CD31的表达、低密度脂蛋白的摄取和细胞的表型;扫描电子显微镜检查细胞表面的微绒毛和窗孔样结构;RT-PCR和免疫细胞化学法检测细胞内源性血管内皮细胞生长因子(VEGF)的表达,并比较各种不同器官微血管内皮细胞对肾上腺糖皮质激素诱导细胞毒的敏感性。结果人肾上腺微血管内皮细胞表达经典内皮细胞标志vWF和CD31,并摄取低密度脂蛋白;同时表达α-1抗胰蛋白酶、肿瘤坏死因子受体(TNFR)p55和细胞间黏附分子-1。扫描电子显微镜显示细胞表面存在大量微绒毛和窗孔样结构。RT-PCR显示人肾上腺微血管内皮细胞高表达VEGF mRNA,免疫细胞化学法显示其细胞内高表达VEGF。对肾上腺皮质激素诱导的细胞毒性反应中,人脑微血管内皮细胞最敏感,人肺和肝窦内皮细胞次之,而人肾上腺微血管内皮细胞表现高度抵抗。结论人肾上腺微血管内皮细胞是特殊分化的,具有不同于其他器官组织内皮细胞的表型和功能特性。
Objective To study the phenotype and functional characteristics of human adrenal microvascular endothelial cells. Methods Human adrenal microvascular endothelial cells were isolated and purified by subcellular cloning. The expression of vWF and CD31, the uptake of LDL and the phenotype of endothelial cells were detected by flow cytometry. Microvilli and window-like structures on the cell surface were examined by electron microscopy. The expression of VEGF was detected by RT-PCR and immunocytochemistry. The effects of adrenoglycan on endothelial cells Corticosteroid-induced cytotoxicity. Results Human adrenal microvascular endothelial cells expressed classical endothelial cell markers vWF and CD31 and uptake of low-density lipoprotein. At the same time, α-1 antitrypsin, tumor necrosis factor receptor (TNFR) p55 and intercellular adhesion molecule-1 were expressed. Scanning electron microscopy revealed a large number of microvilli and window-like structures on the cell surface. RT-PCR showed that human adrenal microvascular endothelial cells express high levels of VEGF mRNA, immunocytochemistry showed high intracellular VEGF expression. In cortico-hormone-induced cytotoxic responses, human brain microvascular endothelial cells are the most sensitive, followed by human lung and sinusoidal endothelial cells, while human adrenal microvascular endothelial cells are highly resistant. Conclusion Human adrenal microvascular endothelial cells are differentiated and have phenotypic and functional properties that are different from endothelial cells in other organs.