香烟烟雾提取物对人内皮细胞细胞间黏附分子-1和白介素-8表达以及与多形核中性粒细胞黏附的影响

Effects of Cigarette Smoking Extract on Expressions of ICAM-1 and IL-8 in Human Endothelial Cells and Adherences of PMN to Human Endothelial Cells

目的探讨香烟烟雾提取物(CSE)诱导作用对人内皮细胞细胞间黏附分子-1(ICAM-I)和白介素-8(IL-8)表达以及与多形核中性粒细胞(PMN)黏附的影响。方法培养人脐静脉内皮细胞株(ECV-304);制备香烟烟雾提取物(CSE);用不同浓度(0、2.5%、5.0%、10.0%)CSE刺激内皮细胞,酶联免疫吸附测定(ELISA)法检测不同时间点(1h、3h、6h、9h、12h、24h、48h)收集的细胞培养上清液IL-8的浓度,细胞免疫化学染色(SABC)检测ICAM-1在不同时间点(3h、6h、9h、12h)内皮细胞上的表达,细胞记数法测定(1h、3h、6h、9h、12h)后PMN与内皮细胞的黏附率(PMN-EC)。结果(1)各CSE浓度组干预12h后,ICAM-1在EC上的表达量随着干预浓度的增加而增高,除了2.5%CSE干预组和5.0%CSE干预组之间无差异外,各组间差异均有非常显著性意义(P<0.01);10.0%CSE干预不同时间点后,随着干预时间的延长,ICAM-1在内皮细胞上的表达量增加,各时间组之间差异均有非常显著性意义(P<0.01)。(2)随着CSE干预浓度的增加和干预时间的延长,内皮细胞表达IL-8的浓度增高,各组间差异均有非常显著性意义(P<0.01)。(3)各CSE浓度组干预12h后,PMN-EC黏附率随着干预浓度的增加而增加,各组间差异均有非常显著性意义(P<0.01)。10.0%CSE干预不同时间点后,随着干预时间的延长,PMN-EC黏附率增加,各时间组差异均有非常显著性意义(P<0.01)。结论香烟烟雾提取物可以刺激激活血管内皮细胞ICAM-1和IL-8表达,增强PMN与内皮细胞的黏附。

Objective To investigate the effects of induction of cigarette smoking extract （CSE） on expressions of intercellular adhesion molecule - 1 （ ICAM - 1 ） and interleukin - 8 （ IL - 8 ） in human endothelial cells, as well as on adherences of polymorphonuclear neutrophils （PMN） to human endothelial ceils. Methods Culture human umbilical vein endothelial cells （ ECV -304） ; Formulate CSE; When ECV -304 is excited by different concentrated CSE as 0%, 2. 5%, 5% and 10%, the concentrations of IL -8 in supematant of culture medium were examined by the methods of ELISA at various time （ 1 h, 3h, 6h, 9h, 12h, 24h, 48h）, the expressions of ICAM - 1 in Ecv -304 at various time （3h, 6h, 9h, 12h） were detected by immunocytochemical stain （SABC）, the adhesive rates of PMN to ECV -304 after different period （lh, 3h, 6h, 9h, 12h） were detected by calculating cell＇ number. Results （ 1 ） After induced ECV -304 by various concentrated of CSE 12 hours, the quantities of expressions of ICAM - 1 in ECV - 304 increased by CSE concentrated increased, the more concentrated the more positive it was stained （ P 〈0.01 ）, except 2. 5% CSE with 5% CSE group （ P 〉0. 05）. Induced ECV - 304 by 10% CSE at various time （0h, 3h, 6h, 9h, 12h）, the quantities of expressions of ICAM -1 in ECV -304 increased by induced time increased. Contrasted the groups at various time （ P 〈 0.05）. （2） Significant increase in IL - 8 concentration （ P 〈 0. 01 ） among all CSE intervened groups ; Reliance on time and concentration intervened of IL - 8 concentration among all groups. （3） After induced ECV - 304 by various concentrated of CSE 12 hours, the adhesive rates of PMN to ECV - 304 increased by CSE concentrated increased, The adhesive rates increased significantly among all groups （ P 〈 0. 01 ）. Induced ECV - 304 by 10% CSE at various time （1h, 3h, 6h, 9h, 12h）, the adhesive rates increased by induced time increased. Contrasted the groups at various time （ P 〈 0.01）. Conclusion CSE could increase the expressions of ICAM - 1 and IL - 8, promote the adherence of PMN to endothelial cells.