华支睾吸虫排泄分泌抗原诱导RAW264.7细胞IL-33表达的研究

Expression of IL-33 in RAW264.7 cells induced by excretory-secretory antigens of Clonorchis sinensis

目的探讨华支睾吸虫排泄分泌抗原(ESA)对巨噬细胞系RAW264.7白介素33(IL-33)表达的影响,并探讨其机制。方法分别在ESA(60μg/ml)与RAW264.7细胞作用2、4、8、12h时检测IL-33 mRNA表达水平。用抗TNF-α中和抗体阻断TNF-α作用,通过RT-PCR及Western blot检测对ESA诱导RAW264.7细胞IL-33表达变化的影响。结果 ESA诱导RAW264.7细胞IL-33mRNA水平增高(P<0.05);利用TNF-α中和抗体阻断TNF-α作用,可显著降低IL-33mRNA的含量(P<0.05)。但ESA对IL-33蛋白水平的表达无显著影响(P>0.05)。结论 ESA可诱导RAW264.7细胞IL-33mRNA高表达,该作用部分依赖于自分泌的炎性因子TNF-α作用。

Objective To investigate the effect of excretory-secretory antigens（ESA）of Clonorchis sinensis on the expression of interleukin-33（IL-33）in the RAW264.7cell line and to explore the preliminary mechanisms of that expression. Methods Real-time PCR（RT-PCR）was used to measure levels of IL-33 mRNA in RAW264.7cells 2,4,8,and12 hafter stimulation with ESA.TNF-α-neutralizing antibody was used to examine the effect on TNF-αin RAW264.7cells treated with ESA,and changes in the level of IL-33 expression were examined using Western blotting and RT-PCR.Results The level of IL-33 mRNA in RAW264.7cells increased after stimulation with ESA（P〈0.05）and this effect was partially reversed by TNF-α-neutralizing antibody（P〈0.001）.The level of IL-33 protein expression tended to increase after stimulation with ESA,but the increase was not significant（P〉0.05）. Conclusion ESA can up-regulate the level of IL-33 mRNA expression in RAW264.7cells and this effect partly depends on the role of the autocrine TNF-α.