人肾上腺微血管内皮细胞的形态、表型和功能特性  

Characteristics of Human Adrenal Microvascular Endothelial Cells in Morphology,Phenotypes,and Function

作　　者：顾锋 张文健[2] 吴练秋[2] 叶丽亚[2] 李汉忠 李静[2] 许世清[2] 徐亚平[2] 娄晋宁[2] 

机构地区：[1]Department of Endocrinology, PUMC Hospital,CAMS and PUMC,Beijing 100730 [2]中日友好医院临床医学研究所 [3]Department of Urology, PUMC Hospital,CAMS and PUMC, Beijing100730

出　　处：《中国医学科学院学报》2006年第4期502-506,i0001,共6页Acta Academiae Medicinae Sinicae

基　　金：国家自然科学基金(30230150)~~

摘　　要：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 investigate the phenotypic and functional characteristics of human adrenal microvascular endothelial cells （AdrEC）. Methods AdrEC were isolated and purified from a sample of human adrenal tissue by sub-cell clone method. The cells identified by flow cytometry for classical endothelial markers yon Willebrand factor （vWF） and CD31, uptake of Dil-labeled acetylated low density lipoprotein （Dil-Ac-LDL）, as well as phenotypes. The cell fenestrations were checked by scanning electron microscopy. The expressions of endogenous vascular endothelial growth factor （VEGF） mRNA and protein were detected by reverse transcription-polymerase chain reaction （RT-PCR） and immunocytochemistry. The glucocorticoid-in- duced cytotoxicities in different organs-derived microvascular endothelial cells were compared. Results Human AdrEC expressed those classical endothelial markers such as vWF, CD31, and uptake of Dil-Ac-LDL. The phenotypic analysis indicated that α-1 proteinase inhibitor, tumor necrosis factor receptor p55, and intercellular adhesion molecule-1 were expressed in human AdrEC. Scanning electron microscopy demonstrated that there were many microviUi and fenestrations on cellular surface. RT-PCR and immunocytochemistry showed that there was expression of endogenous VEGF in AdrEC. In response to glucocorticoid-induced cytotoxicity, microvascular endothelial cells （MVEC） derived from human brain were highly susceptible, MVEC derived from hu- man lung and human liver sinusoidal endothelial cells were sub-sensitive, while AdrEC were highly resistant. Conclusion Human AdrEC are specially differentiated and have characteristics that are different from other organ-derived MVEC in phenotypes and functions.

关 键 词：肾上腺 内皮细胞 血管内皮生长因子 细胞毒 

分 类 号：R331.32[医药卫生—人体生理学]