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[人肾上腺微血管内皮细胞的形态、表型及功能特征]

[Characteristics of human adrenal microvascular endothelial cells in morphology, phenotypes, and function].

作者信息

Gu Feng, Zhang Wen-Jian, Wu Lian-Qiu, Ye Li-Ya, Li Han-Zhong, Li Jing, Xu Shi-Qing, Xu Ya-Ping, Lou Jin-Ning

机构信息

Institute of Clinical Medical Sciences, China-Japan Friendship Hospital, Beijing 100029, China.

出版信息

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2006 Aug;28(4):502-6.

Abstract

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 von 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-induced 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 alpha-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 microvilli 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 human 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.

摘要

目的

研究人肾上腺微血管内皮细胞(AdrEC)的表型和功能特征。

方法

采用亚细胞克隆法从人肾上腺组织样本中分离纯化AdrEC。通过流式细胞术鉴定细胞的经典内皮标志物血管性血友病因子(vWF)和CD31、Dil标记的乙酰化低密度脂蛋白(Dil-Ac-LDL)摄取情况以及表型。通过扫描电子显微镜检查细胞窗孔。采用逆转录-聚合酶链反应(RT-PCR)和免疫细胞化学检测内源性血管内皮生长因子(VEGF)mRNA和蛋白的表达。比较不同器官来源的微血管内皮细胞中糖皮质激素诱导的细胞毒性。

结果

人AdrEC表达vWF、CD31等经典内皮标志物,并摄取Dil-Ac-LDL。表型分析表明,人AdrEC表达α-1蛋白酶抑制剂、肿瘤坏死因子受体p55和细胞间黏附分子-1。扫描电子显微镜显示细胞表面有许多微绒毛和窗孔。RT-PCR和免疫细胞化学显示AdrEC中有内源性VEGF表达。在糖皮质激素诱导的细胞毒性反应中,人脑血管来源的微血管内皮细胞(MVEC)高度敏感,人肺和人肝窦状内皮细胞来源的MVEC中度敏感,而AdrEC高度耐药。

结论

人AdrEC具有特殊分化,在表型和功能上与其他器官来源的MVEC不同。

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