TNF-α对人脐静脉内皮细胞表达MCP-1及IL-8的影响

Effects of TNF-α on MCP-1 and IL-8 mRNA Expression in Human Umbilical Vein Endothelial Cells

本研究探讨肿瘤坏死因子α(TNF-α)刺激人脐静脉内皮细胞(HUVEC)后对细胞表达单核细胞趋化蛋白-1(MCP-1)和白介素-8(IL-8)的影响及其可能的分子机制。用RT-PCR的方法检测MCP-1和IL-8mRNA的表达,免疫荧光染色法检测HUVEC核内转录因子κB(NF-κBp65)的激活。结果表明:TNF-α刺激HUVEC后,细胞内MCP-1和IL-8mRNA的表达增强,且有时相性变化;IL-8mRNA表达8小时达到高峰,MCP-1mRNA表达12小时达到高峰。细胞核内NF-κBp65蛋白表达增强,在感染后0.5小时开始增加,1小时达峰值。然后,胞浆染色逐渐减弱,而核染色增强,表明转录因子NF-κBp65明显的核移位。结论:TNF-α刺激HUVEC后能增加细胞内MCP-1、IL-8mRNA和NF-κBp65蛋白质的表达,提示TNF-α可能通过NF-κB途径诱导血管内皮细胞MCP-1和IL-8等炎症介子的过度表达。

This study was aimed to explore monocyte chemotactic protein 1 （MCP-1） and interleukin 8 （IL-8） expressions in human umbilical vein endothelial cells （HUVECs）affected by TNF-α and its molecular mechanism. RT- PCR assay was used to detect the expression of MCP-1 and IL-8 mRNA in HUVECs at various times following stimulation with TNF-α; the nuclear factor-kappa B （NF-KB） activation in HUVECs were detected by immunofluo- rescence. The results showed that after stimulation with NF-κB , the expression levels of MCP-1 and IL-8 mRNA in HUVECs increased, and changed along with different time; the expression of IL-8 mRNA reached to peak level at 8 hours and the expression of MCP-1 mRNA reached to peak level at/2 hours; the expression of NF-KB p65 protein began increasing at 30 minutes and reached to peak level at 1 hour; then the staining of cytoplasm gradually weakened, while staining of nuclei was enhanced, which indicated significant nuclear translocation of NF-KB p65. It is concluded that the TNF-et induces expressions of IL-8 mRNA, MCP-1 mRNA and NF-KB in HUVECs, and NF-KB activitied signal pathway may play a role in IL-8 mRNA and MCP-1 mRNA expressions.