摘要
目的观察肿瘤坏死因子α(tumour necrosis factor alpha,TNF-α)对人脐静脉内皮细胞(HUVECs)凋亡及炎症反应的影响,并探讨其损伤血管内皮细胞的可能机制。方法常规培养的HUVECs至第三代,随机分为对照组和TNF-α组。对照组常规方法培养作为对照,TNF-α组应用200 ng/ml的TNF-α刺激HUVECs 6 h后,采用流式细胞法检测HUVECs的凋亡率,Western blot法检测ICAM-1、VCAM-1、E-selectin和eNOS的蛋白表达水平;流式细胞术和Western blot法进一步评价核因子-κB(NF-κB)、激活蛋白-1(AP-1)的表达激活情况。结果与对照组相比,200 ng/ml TNF-α能明显增加HUVECs的凋亡率(P<0.05),同时上调ICAM-1、VCAM-1、E-selectin炎症因子在HUVECs中的表达(P<0.05),TNF-α抑制eNOS的释放(P<0.05),并激活NF-κB、AP-1的表达(P<0.05)。结论 TNF-α能显著增加HUVECs的凋亡和炎症因子表达,其损伤机制可能是通过激活NF-κB、AP-1途径,并抑制eNOS的释放有关。
Objective To observe the effect of tumor necrosis factor-alpha（TNF-ct） on apoptosis and inflammation of hu- man umbilical vein endothelial cells（ HUVECs ） and investigate the possible mechanism. Methods Cultured HUVECs were randomly divided into control group and TNF-α group. After pretreated with TNF-α for 6 hours, the apoptosis rate was tested by flow cytometry, and the expression of ICAM-1, VCAM-1, E-selectin and eNOS were detected by Western blot assay. The activation of NF-KB and AP-1 were detected by flow eytometry and Western blot. Results Compared with the control group, TNF-α significantly increased the apoptosis rate of HUVECs （ P 〈 0.05 ） and the expression of ICAM-1, VCAM-1, E-selectin in HUVECs（P 〈 0.05） and inhibited the release of eNOS（P 〈 0.05 ）. At the same time, TNF-α activated the expressions of NF-KB and AP-1 （ P 〈 0.05 ）. Conclusion TNF-oL significantly increases the apopto- sis and inflammation of HUVECs. The mechanism of this injury might be related to the activation of the NF-KB and AP- 1 signaling pathway and inhibiting the release of eNOS.
作者
陈铁龙
祝光礼
张旭栋
孟伟康
汪禹
CHEN Tie-long;ZHU Guang-li;ZHANG Xu-dong;et al(Department of Cardiology, Hangzhou Hoapital of Traditional Chinese Medicine, Hangzhou, Zhejiang 310007, China)
出处
《中华全科医学》
2018年第2期184-187,共4页
Chinese Journal of General Practice
基金
浙江省中医药科技计划项目(2008-CBO54)
浙江省杭州市科技局医院重点专科专病项目(20160533-B63,20150733Q57)
杭州市卫生科技计划重点项目(2017Z10)
