CpG-ODN对鼠巨噬细胞TLR-9及Th1／Th2类细胞因子分泌的影响  

Effects of CpG-ODN on TLR-9 expression and Th1/Th2 cytokines secretion from mouse macrophages

作　　者：黄维琳[1] 王炯[1] 刘荣玉[1] 

机构地区：[1]安徽医科大学第一附属医院干部呼吸内科

出　　处：《安徽医科大学学报》2008年第1期28-31,共4页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金资助项目(编号:30670936)

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RAW264.7Toll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(IL-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-9 mRNA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IL-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。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.

关 键 词：寡脱氧核糖核苷酸类/药理学 巨噬细胞/药物作用 巨噬细胞/细胞学 TH1细胞 Th2细胞 细胞因子类 氯喹 

分 类 号：R392.11[医药卫生—免疫学] R342.4[医药卫生—基础医学]