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目的探讨干扰素(IFN)-γ对内皮细胞上Toll样受体4(toll like receptor-4,TLR4)和髓样分化蛋白-2(myeloid differentiation protein-2,MD-2)表达的影响以及在内毒素(LPS)对内皮细胞激活中的影响。方法分离、培养人脐静脉内皮细胞,逆转录-聚合酶链反应(RT-PCR)和Western blot方法检测IFN-γ刺激后内皮细胞TLR4、MD-2表达的变化;TransAM法检测核因子(NF)-кB活性,酶联免疫吸附分析(ELISA)法检测IL-8水平。结果IFN-γ能明显上调内皮细胞TLR4、MD-2的表达,并且TLR4、MD-2表达的增加与IFN-γ呈一定的剂量依赖关系;IFN-γ对内皮细胞NF-кB的活性无明显影响,但用IFN-γ孵育后的内皮细胞却对LPS的反应明显增强,NF-кB活性明显增高,白细胞介素(IL)-8分泌明显增加,且与IFN-γ呈剂量依赖关系。结论IFN-γ通过上调内皮细胞上的LPS受体-TLR4、MD-2而促进LPS对内皮细胞的活化。
Objective To investigate the effect of interferon (IFN) -γ on the expression of toll like receptor 4 (TLR4) and myeloid differentiation protein 2 (MD-2) on endothelial cells, Effects of Endotoxin (LPS) on Endothelial Cell Activation. Methods Human umbilical vein endothelial cells were isolated and cultured. The changes of TLR4 and MD-2 expression in endothelial cells were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The expressions of nuclear factor ) -kappaB activity and IL-8 levels by enzyme-linked immunosorbent assay (ELISA). Results IFN-γ significantly up-regulated the expression of TLR4 and MD-2 in endothelial cells, and the increase of TLR4 and MD-2 expression was in a dose-dependent manner with IFN-γ. IFN-γ showed no significant effect on NF-κB activity in endothelial cells However, the endothelial cells incubated with IFN-γ had a significantly increased response to LPS, a marked increase in NF-κB activity, a significant increase in interleukin-8 secretion, and a dose-dependent relationship with IFN-γ. Conclusion IFN-γ promotes the activation of endothelial cells by LPS by up-regulating the LPS receptors TLR4 and MD-2 on endothelial cells.