呼吸道合胞病毒感染巨噬细胞诱导COX-2表达的相关研究

The relative study of the expression of COX-2 in respiratory syncytial virus-infected macrophages

目的探讨呼吸道合胞病毒(respiratory syncytial virus,RSV)感染肺巨噬细胞RAW264.7细胞时,环氧化酶-2(cyclooxygen-ase-2,COX-2)的表达及其相关炎症介质PGE2(Prostaglandin E2)的水平变化,初步阐明COX-2在RSV感染所致炎性反应中的作用,为研究RSV感染时的炎症产生机理,临床治疗RSV感染所致的炎症反应等提供一定的理论依据。方法RSV感染体外培养的RAW264.7巨噬细胞,分别于不同感染时间点(4、8、16、24 h)收集细胞和细胞培养上清。用半定量逆转录聚合酶链反应(RT-PCR)和Western-blot法分别检测细胞中环氧化酶-2(COX-2)的mRNA和蛋白表达,酶联免疫法(ELISA)检测细胞培养上清PGE2含量变化。结果RSV感染8 h后即上调RAW264.7巨噬细胞COX-2 mRNA转录和蛋白水平的表达。细胞培养上清中PGE2的含量逐渐增加。各指标的变化与正常对照相比差异均有显著性,并且随着RSV感染时间的延长而表达量增加。结论RSV感染RAW264.7巨噬细胞早期即可诱导COX-2的表达,同时生成大量的PGE2,PGE2的合成主要受COX-2的调节,COX-2参与了RSV感染所致的炎症反应。

Aim To investigate the effect of expression of Cyclooxygenase-2 （COX-2） and its related inflammatory mediators on RAW264.7 macrophages infected with respiratory syncytial virus （RSV） in vitro, and to provide theoretical foundation for the mechanism of RSV induced inflammatory response and clinical treatment of RSV disease. Methods RAW264.7 macrophages infected with RSV in vitro were used to collect cells and cellular supernatants. The expression of Cyclooxygenase-2 （ COX - 2） mRNA and protein were evaluated by semiquantitative RT -PCR and Western-blot, respectively. The concentration of Prostaglandin E2 （PGE2） in cellular supernatants was measured by ELISA assay according to the kit reference. Results RSV infection could markedly up-regulate the expression of COX - 2 mRNA and protein in RAW264.7 macrophages since 8 h after infection. In the cellular supernatants, the levels of PGE2 rose gradu- ally. Meanwhile, the changes of each index varied significantly compared with the control group, increased obviously in a time-dependent manner. Conclusions RSV infection macrophages could induce the expression of COX-2 in early stage, COX-2 involved in the inflammatory response of RSV infection.