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H2O2促进中性粒细胞对人脐静脉血管内皮细胞的黏附和损伤

钱中清^*, 何小燕, 吕静竹, 汪洪涛, 方芳

蚌埠医学院免疫学教研室，安徽省感染免疫重点实验室，蚌埠 233003

摘要

本文旨在探讨H₂O₂对中性粒细胞(polymorphonuclear neutrophils, PMNs) 黏附和损伤人脐静脉血管内皮细胞(human umbilical vein endothelial cells, HUVECs)的促进作用。应用相差显微镜和扫描电镜，以未经H2O2处理的PMNs与HUVECs共培养为对照，观察H₂O₂预处理的PMNs对HUVECs的黏附和损伤，并计数其黏附率；应用流式细胞仪检测H₂O₂ 预处理的PMNs对HUVECs表达细胞间黏附分子1 (intercellular adhesion molecule 1, ICAM-1)和Apo2.7水平的影响。结果显示，外源性H₂O₂预处理的PMNs与HUVECs共培养后，相差显微镜下可见圆形PMNs黏附于HUVECs表面，HUVECs细胞形态呈圆形或椭圆形，体积变小；扫描电镜下，细胞微绒毛变短或消失，部分细胞胞膜结构破坏，甚至有细胞碎片形成；H₂O₂预处理的PMNs对HUVECs的黏附率为(57.74 ± 9.18)%，显著高于对照组[(23.12 ± 6.43)%, P < 0.01, n = 8]；HUVECs表达黏附分子ICAM-1及凋亡相关分子Apo2.7的百分率分别为(44.69 ± 1.52)%和(39.29 ± 1.81)% ，显著高于对照组[(21.79 ± 1.43)%和(9.79 ± 1.43)% ](P < 0.01, n = 8)。以上结果表明H₂O₂可促进PMNs对HUVECs的黏附和损伤。

关键词： 过氧化氢; 中性粒细胞; 血管内皮细胞; 细胞间黏附分子1

分类号：R331.1

[H2O2 promotes neutrophil adherence and injury of human umbilical vein endothelial cells.] [Article in Chinese]

QIAN Zhong-Qing^*, HE Xiao-Yan, LV Jing-Zhu, WANG Hong-Tao, FANG Fang

Key Laboratory of Anhui Province for Infection and Immunology, Department of Immunology, Bengbu Medical College, Bengbu 233003, China

Abstract

To explore the role of hydrogen peroxide (H2O2) in promoting polymorphonuclear neutrophils adherence and injury of human umbilical vein endothelial cells (HUVECs), the ordinary optical microscope and scanning electron microscopy were used to observe the adherence and injury after HUVECs co-cultured with neutrophils pretreated by extracellular H2O2 (HUVECs and neutrophils co-culture without H2O2 pretreatment as control), and the adhesion rates of neutrophils were measured through cell count test. The percentages of HUVECs expressing intercellular adhesion molecule 1 (ICAM-1) and Apo2.7 were detected by flow cytometry. After being cocultured with the neutrophils pretreated by extracellular H2O2, HUVECs showed obvious injury changes, such as round or oval shape, shortened or disappeared microvilli, and membrane structural damage; The adhesion rate of neutrophils was (57.74 ± 9.18)%, which was significantly higher than that in control [(23.12 ± 6.43)%, P < 0.01, n = 8]; The percentages of HUVECs expressing ICAM-1 and Apo2.7 were (44.69 ± 1.52)% and (39.29 ± 1.81)% respectively, which were significantly higher than those in control [(21.79 ± 1.43)% and (9.79 ± 1.43)%] (P < 0.01, n = 8). The results suggest that extracellular H2O2 can promote the neutrophils adherence and injury of HUVECs.

Key words: Hydrogen peroxide; neutrophil; human umbilical vein endothelial cell; intercellular adhesion molecule 1

收稿日期：2012-01-05　　录用日期：2012-04-20

通讯作者：钱中清　　E-mail: qzq7778@yahoo.com.cn

引用本文：

钱中清, 何小燕, 吕静竹, 汪洪涛, 方芳. H2O2促进中性粒细胞对人脐静脉血管内皮细胞的黏附和损伤[J]. 生理学报 2012; 64 (4): 412-416.

QIAN Zhong-Qing, HE Xiao-Yan, LV Jing-Zhu, WANG Hong-Tao, FANG Fang. [H2O2 promotes neutrophil adherence and injury of human umbilical vein endothelial cells.] [Article in Chinese] . Acta Physiol Sin 2012; 64 (4): 412-416 (in Chinese with English abstract).