炎症细胞因子对人脐静脉内皮细胞分泌VEGF、ICAM-1的影响

Effects of inflammatory cytokine on secretions of VEGF and ICAM-1 in human umbilical vein endothelial cells

目的:探讨炎症因子白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNFα)对人脐静脉内皮细胞(HU-VEC)分泌血管内皮生长因子(VEGF)、细胞间粘附分子1(ICAM-1)的影响。方法:人脐静脉内皮细胞在37℃、5%CO2的条件下常规培养,实验采用4～5代细胞;按1×105/ml密度接种于5 cm2的培养皿中,24 h后换液,生长接近80%融合时进行干预。按IL-1β组、TNF-α组、IL-1β+TNF-α组、空白对照组进行刺激(IL-1β和TNFα干预浓度均为10 ng/ml),每组12个样本;在共同培养24 h后收集细胞上清液。应用酶联免疫吸附测定(ELISA)法检测细胞上清液中VEGF、ICAM-1的浓度。结果:IL-1β组、TNF-α组、IL-1β+TNF-α组的VEGF值(ng/ml)分别为(27.91±0.63)、(13.54±1.17)和(13.05±0.11),均明显高于对照组(4.21±0.91),P=0.000;IL-1β组、TNF-α组、IL-1β+TNF-α组的ICAM-1值(ng/ml)分别为(2.88±0.98)、(2.96±0.03)和(2.30±0.20),均明显高于空白对照组(0.25±0.01),P=0.000。结论:炎症因子IL-1β、TNF-α均可促进人脐静脉内皮细胞分泌斑块不稳定相关标志物VEGF、ICAM-1,可能是炎症在急性冠脉综合征的发生、发展中起重要作用的机制之一。

Objective： To observe the effect of interleukin-1β （IL-1β） and tumor neerosis faetor-α （TNF-α） on secretions of vascular endothelial growth factor （VEGF） and intercellular adhesion moleeule--1 （ICAM--1） in human umbilical vein endothelial cell （HUVEC）. Methods： HUVECS were incubated in a 37℃, 5% CO2 air. Cells of the fourth to fifth passage were used for experiments, seeded at 1 × 10^5 cells/ml per 5 cm^2 tissue culture flasks; change the medium 24 h after establishing the subculture. When cells reached 80 % confluence, the following treatment would be done： After 24 h exposure to different inflammatory eytokines, serum VEGF and ICAM-1 were determined by enzyme-linked immunosorbent assay （ELISA） in control group, IL-1β group, TNF- αgroup, IL -1β＋ TNF-α group. The concentration of IL--1β and TNF-α was 10 ng/ml. Results： ELISA analysis showed that the concentration of VEGF in IL-1β, TNF-α and IL-1β＋TNF-α groups were （27.91±0. 63）, （13.54±1.17） and （13.05± 0.11） ng/ml respectively; whereas the concentration of VEGF of the control group was （4. 21±0. 91） ng/ml. There were significant differences between all the experimental groups and the control group （P= 0. 000）. The concentration of ICAM--1 in IL-1β, TNF-α and IL-1β＋TNF--α groups were （2.88±0. 98）, （2. 96±0. 03） and （2. 30±0. 20） ng/ml respectively; whereas the concentration of ICAM- 1 of the control group was （0. 25 ±0.01） ng/ml. There were significant differences between all the experimental groups and the control group （P=0. 000）. Conclusion： IL-1β and TNF-α can induce the HUVECS production of VEGF and ICAM- 1, suggesting that inflammation may play an important role in the development of acute coronary syndromes.