【摘要】 目的:观察香烟提取物(CES)对人脐静脉内皮细胞的影响,并初步探讨吸烟导致动脉粥样硬化的可能机制。方法:采用浓度分别为2.5%、5%、10%、20%和40%的CSE以相同的时间24h处理人脐静脉内皮细胞,另用同一浓度10%CSE分别处理人脐静脉内皮细胞6,12,24和48 h,用MT方法分析浓度及时间梯度CES处理后细胞的抑制率。采用未加CES处理的人脐静脉内皮细胞作为对照组,ELISA方法检测VCAM-1、ICAM-1、TNF-α及IL-6等细胞因子的水平;Western blot方法检测人脐静脉内皮细胞自噬基因蛋白的表达水平。结果:MTT检测显示CSE以浓度梯度和时间梯度的方式降低HUVEC的细胞活性;ELISA检测结果表明CES增加VCAM-1、ICAM-1、TNFα及IL-6等黏附因子和炎症因子水平;Western blot结果表明10%CSE处理人脐静脉内皮细胞会降低抑制凋亡基因Bcl-2蛋白的表达水平,增加促凋亡基因Bax及促进细胞自噬的基因Beclin1的蛋白表达水平。结论:研究结果显示CES可能通过提高细胞黏附因子和炎症因子水平,促进HUVEC的凋亡和自噬,从而对内皮细胞及血管内皮产生影响。更多还原

【Abstract】 Objective: To observe the influence of cigarette smoking extract( CES) on human umbilical vein endothelial cells( HUVECs),and to explore the possible mechanism of atherosclerosis caused by smoking. Methods: HUVEC was treated for 24 hours at CSE concentration of 2. 5%,5%,10%,20% and 40%respectively,Then,HUVEC was treated by 10% CES for 6,12,24 and 48 hours respectively,MTT method was used to detect cell inhibition by CES. Control group was CES-free,ELISA was used to measure VCAM-1,ICAM-1,TNF-α,and IL-6 cytokine level; and Western blot was used to detect the HUVEC autophagy gene protein expression level. Results: MTT assay method showed that CSE reduce cell viability of HUVEC in concentration and time gradient.; ELISA test results showed that CSE significantly increased cytokines like VCAM-1,ICAM-1,TNFα,IL-6; Western blot results showed that 10% CES treatment group will decrease the protein expression level of apoptosis-inhibition gene Bcl-2 and increase the protein expression level of apoptosis-promotion gene Bax and autophagy-promotion gene Beclin1. Conclusion: The results showed that CES may increase the level of adherence and inflammatory cytokines,promote HUVEC apoptosis and autophagy,resulting in deterioration of vascular endothelial function.更多还原