中文摘要：

目的探讨非甲基化的胞嘧啶-磷酸-鸟嘌呤二核苷酸序列（CpG—ODN）对鼠单核-巨噬细胞系RAW264.7 Toll样受体9（TLR-9）的表达及Th1／Th2类细胞因子分泌的影响。方法体外培养小鼠单核-巨噬细胞系RAW264．7细胞，逆转录聚合酶链反应法（RT-PCR）及Western blot检测Cp-ODN，鸟嘌呤胞嘧啶二核苷酸序列（Non—CpG—ODN），CpG—ODN＋氯喹刺激24h后巨噬细胞TLR-9的表达，放免法及ELISA法检测刺激前后巨噬细胞上清液中肿瘤坏死因子-α（TNF-α）、白细胞介素-6（1L-6）及白细胞介素-12（IL-12）的表达水平。结果刺激24h后，CpG—ODN与CpG—ODN＋氯喹组巨噬细胞TLR-9mRNA及蛋白的表达明显增加（P〈0．01，P〈0．05）；细胞上清液中TNF-α水平CpG—ODN组明显高于其他组，差异有显著性（P〈0．05）；各组1L-6、IL-12水平无显著性差异（P〉0．05）。结论CpG—ODN刺激可引起鼠巨噬细胞TLR-9表达增高，调节Th1／Th2类细胞因子的分泌。

英文摘要：

Objective To investigate the effects on toll like receptor-9 （TLR-9）expression and Th1/Th2 cytokines secretion from mouse macrophages （ RAW264. 7 ） induced by the synthetic oligodeoxynucleoties containing unmethylated CpG motifs （CpG-ODN）. Methods RAW264.7 cells were grown in vitro. Stimulated by CpG-ODN, synthetic oligodeoxynucleoties containing GpC motifs （Non-CpG-ODN） or CpG-ODN ＋ chloroquine for 24 hours, then the expressions of TLR-9 mRNA and protein were analyzed by RT-PCR and Western blot respectively. The concentration of tumor necrosis factor-α（TNF-α）and interleukin-6 （IL-6） in the supernatants of each group were examined by Radioimmunoasssy（RIA） kits and the level of interleukin-12 （IL-12） was analyzed by ELISA. Results After treated for 24 hours, the expressions of TLR-9 mRNA and TLR-9 protein in CpG-ODN group and CpG-ODN ＋ chloroquine group were significantly higher than that in other groups. The level of TNF-α in the supernatant in the CpG- ODN group was higher than that in other groups. The difference between these two groups was significant （P 〈 0. 05）. The levels of IL-6 and IL-12 in the supernatants had no significant difference between these groups（ P 〉 0. 05）. Conclusions CpG-ODN can enhance the expressions of TLR-9 and adjust the expressions of Th1 and Th2 cytokines from macrophages.