The effect of TNF-α on apoptosis and inflammation of human umbilical vein endothelial cells by NF-Κb and AP-1 signal pathway
-
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。Abstract: Objective To observe the effect of tumor necrosis factor-alpha(TNF-α) on apoptosis and inflammation of human 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-κB and AP-1 were detected by flow cytometry 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-κB and AP-1(P<0.05). Conclusion TNF-α significantly increases the apoptosis and inflammation of HUVECs. The mechanism of this injury might be related to the activation of the NF-κB and AP-1 signaling pathway and inhibiting the release of eNOS.
点击查看大图
计量
- 文章访问数: 621
- HTML全文浏览量: 50
- PDF下载量: 58
- 被引次数: 0