天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (7): 687-691.doi: 10.11958/20190304

• 细胞与分子生物学 • 上一篇    下一篇

细胞交互作用在重叠综合征所致血管内皮损伤中及与细胞间黏附分子-1的作用研究

肖凡，曹洁，李鑫   

  1. 1天津市第三中心医院分院呼吸科（邮编 300250）；2天津医科大学总医院呼吸与危重症学科
  • 收稿日期:2019-01-30 修回日期:2019-05-07 出版日期:2019-07-15 发布日期:2019-08-01
  • 通讯作者: 肖凡 E-mail:luciax@sina.com

The importance of cellular interaction in vascular endothelial injury induced by overlap syndrome and the role of intercellular adhesion molecule-1

XIAO Fan，CAO Jie，LI Xin   

  1. 1 Department of Respiratory Medicine, Tianjin Third Central Hospital-Branch Hospital, Tianjin 300250, China; 2 Department of Respiratory and Critical Care, Tianjin Medical University General Hospital
  • Received:2019-01-30 Revised:2019-05-07 Published:2019-07-15 Online:2019-08-01

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摘要： 目的 探讨中性粒细胞（PMN）与血管内皮细胞（VEC）的交互作用在重叠综合征所致血管内皮损伤中所起的作用以及与细胞间黏附分子（ICAM）-1的作用机制。方法 每次取样均抽取 16例健康志愿者空腹外周静脉血，每人 2.5 mL，每次取样共 40 mL，先后共进行 10次取样。使用双层密度梯度离心法分离出 PMN，将 PMN与 VEC直接共培养的实验组与单纯 VEC培养的对照组同时置于细胞培养箱内培养 4 h，应用天津医科大学总医院呼吸与危重症学科呼吸仿真系统进行间歇伴持续低氧暴露 8 h，暴露后分别收集上清液和 VEC。分别检测上清液中白细胞介素（IL）-6、肿瘤坏死因子（TNF）-α、ICAM-1、过氧化氢酶（CAT）活性和丙二醛（MDA）浓度；qRT-PCR检测 VEC促凋亡基因Bak和抑凋亡基因 Bcl-xl mRNA 表达水平。结果 实验组 IL-6、TNF-α、ICAM-1及 MDA的浓度均高于对照组，而CAT的活力值低于对照组（P＜0.05）。实验组 BAK mRNA、Bcl-xl mRNA表达水平均高于对照组，而 Bcl-xl/Bak低于对照组（P＜0.05）。实验组 ICAM-1 与 IL-6、TNF-α、MDA 呈较强的正相关，而与 CAT、Bcl-xl/Bak 呈较强的负相关（P＜0.05）。结论 PMN 与 VEC 的交互作用使重叠综合征所致血管内皮损伤更严重，且 ICAM-1在其中起了重要作用。

关键词: 血管系统损伤, 胞间黏附分子-1, 炎症, 氧化性应激, 细胞凋亡, 细胞交互, 重叠综合征

Abstract: Objective To explore the important role of neutrophil (PMN) and vascular endothelial cell (VEC) interaction in vascular endothelial injury induced by overlap syndrome and the relationship with the action of intercellular adhesion molecule (ICAM)-1 at the cellular level. Methods Samples of fasting peripheral venous blood were taken from 16 healthy volunteers, 2.5 mL per person, 40 mL were sampoed at each time and 10 times were sampoed successively. PMN was separated by double-layer density gradient centrifugation. Then the experimental group directly co-cultured with neutrophils and endothelial cells was placed in the cell culture incubator simultaneously with the control group of endothelial cells alone.The culture was carried out for 4 hours, and then the respiratory simulation system of the Department of Respiratory Medicine of Tianjin Medical University General Hospital was used for intermittent and continuous hypoxia exposure for 8 hours. After exposure, the supernatant and endothelial cells were collected separately. The inflammatory factors in the supernatant including interleukin (IL) - 6, tumor necrosis factor (TNF) - α, adhesion molecule ICAM-1, catalase (CAT)activity and malondialdehyde (MDA) concentration were detected. The expression levels of VEC pro-apoptotic gene Bak and anti-apoptotic gene Bcl-xl mRNA were detected by qRT-PCR. Results The concentrations of IL-6, TNF-α, ICAM-1 and MDA were significantly higher in the experimental group than those in the control group (P＜0.05). The activity of CAT was significantly lower in the experimental group than that in the control group (P＜0.05). The expression levels of Bak mRNA and Bcl-xl mRNA were significantly higher in the experimental group than those in the control group, while Bcl-xl/Bak was significantly lower than that in the control group (P＜0.05). In experimental group ICAM-1 was positively correlated with IL-6, TNF - α and MDA, but ICAM-1 was negatively correlated with CAT and Bcl-xl / BAK (P＜0.05). Conclusion The interaction between PMN and VEC aggravates vascular endothelial injury caused by overlap syndrome, and ICAM-1 plays an important role in it.

Key words: vascular system injuries, intercellular adhesion molecule-1, inflammation, oxidative stress, apoptosis, cell interaction；overlap syndrome