摘要
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-γ （IFN-γ） on the expressions of myeloid differentiation protein-2 （MD-2） and toll like receptor 4 （TLR4） and role of lipopolysaccharide （LPS） in activation of endothelial cells. Methods Based on cultured human umbilical vein endothelial cell （HUVEC）, the effect of IFN-γ on the expression of MD-2 and TLR4 was analyzed with reverse transcription polymerase chain reaction （RT-PCR） and Western blot. NF-κB activity was detected by TransAM and level of IL-8 by ELISA. Results IFN-γ could obviously upregulate the expressions of TLR4 and MD-2 in a dose-dependent manner. IFN-γ exerted insignificant effect on activity of NF-κB, but after the endothelial cells were cultured with IFN - γ, LPS reaction was boosted, NF - κB activity enhanced, secretion of IL - 8 significantly increased, relating with IFN-γ in a dose-dependent manner. Conclusion IFN-γ promotes LPS-induced endothelial cell activation through upregulating the expressions of TLR4 and MD-2.
出处
《中华创伤杂志》
CAS
CSCD
北大核心
2005年第12期938-942,共5页
Chinese Journal of Trauma
基金
国家重点基础研究发展规划资助项目(G1999054203)
