白三烯D_4和白介素13对人肺成纤维细胞白三烯受体1 mRNA表达及嗜酸粒细胞趋化因子产生的影响  

Effect of LTD_4 and IL-13 on the expression of CysLT1R and production of eotaxin in human lung fibroblasts

作　　者：柏宏坚 孙建 何礼贤[2] 揭志军[2] 

机构地区：[1]江苏省盐城市第一人民医院呼吸科,盐城224001 [2]复旦大学呼吸病研究所,上海200032

出　　处：《现代免疫学》2007年第1期32-37,共6页Current Immunology

基　　金：江苏省135工程医学重点人才基金资助项目(RC2003109)

摘　　要：探讨白三烯D4(LTD4)和白介素13(IL-13)对人肺成纤维细胞白三烯受体(CysLT1R)表达和eotaxin产生的影响。采用培养人肺成纤维细胞株(MRC-5),以不同浓度的IL-13进行刺激,用实时定量RT-PCR法测定CysLT1R mRNA表达的变化。再以不同浓度的LTD4和IL-13刺激,用ELISA法测定上清液中eotaxin的浓度,并观察Montelukast对eotaxin的产生是否有抑制作用。结果显示,培养的人肺成纤维细胞上存在着CysLT1R mRNA的低表达,以10 ng/ml IL-13刺激时,随着刺激时间延长,CysLT1R mRNA表达逐渐增高,48 h达高峰,较未刺激时表达增加(18.56±7.41)倍(P<0.01)。当以10 ng/ml和100 ng/ml的IL-13浓度刺激48 h时,较未刺激细胞CysLT1R mRNA表达分别增加(17.33±3.81)倍和(20.11±5.05)倍(P<0.01)。IL-13也能促进成纤维细胞产生eotaxin,以10 ng/ml和100 ng/ml的IL-13刺激成纤维细胞时,eotaxin浓度分别为(155.43±15.95)pg/ml和(221.31±17.23)pg/ml,较未刺激细胞(31.67±3.49)pg/ml明显增加(P<0.01)。单独给予LTD4刺激时,eotaxin浓度则无明显变化。但是以1×10-7mol/L和1×10-6mol/L浓度的LTD4分别联合10 ng/ml的IL-13共同刺激时,产生eotaxin的浓度分别为(204.4±25.4)pg/m和(255.1±38.3)pg/ml;较仅给予10 ng/ml的IL-13刺激时(155.4±16.0)pg/ml明显增加(P<0.01)。在LTD4+IL-13刺激组中,加入Mointelukast后eotaxin浓度为(148.3±15.7)pg/ml,较不加Montelukast(204.4±25.4)pg/ml明显降低(P<0.01)。人肺纤维细胞存在着CysLT1R mRNA的低表达,IL-13能够上调其表达,呈现时间与浓度依赖性。IL-13和LTD4对人肺成纤维细胞分泌的eoitaxin具有协同刺激作用,这可能与IL-13上调成纤维细胞CysLT1R mRNA表达有关,而Montelukast对这种刺激具有拮抗作用。To investigate whether Interleukin-13（IL-13） and leukotriene D4 can enhance the expression of cystein leukotriene 1 receptor （CysLT1R） and the production of eotaxin on human lung fibroblasts. Human fetal lung fibroblasts （MRC-5） were cultured and stimuhaed with different dose of IL-13, then the expression of CysLT1R in MRC-5 was investigated by quantitative real-time PCR. After stimulated by IL-13 or/and LTD4, the concentration of eotaxin in the cell supernatant fluid from MRC-5 were measured eotaxin was observed. It was found that only a low level of CysLT1R mRNA was expressed in unstimulated MRC-5, in contrast, stimulated with IL-13 at a concentration of 10 ng/ml fro 48 h, the expression CysLT1R mRNA was 18.56±7.41 times higher than that of unstimulated cells. When stimulated with 100 ng/ml IL-13, it was 20.11±5.05 times higher than unstimulated cells. IL-13 also enhanced the production of eotaxin, in which stimulated with 10 ng/ml or 100 ng· ml IL-13, the concentration of eotaxin produced by MRC-5 was （155.43±15.95） pg/ml or （221.31±17.23） pg/ml, which were significantly higher than that of unstimulated cell （P〈0.01）. LTD4 singly stimulating MRC-5 didn＇t enhance the production of eotaxin. However when 10^-7mol/L or 10^-6 mol/L LTD4 and 10 ng/ml IL-13 together stimulated MRC-5, the concentration of eotaxin was （204.4±25.4） pg/ml or （255.1±38.3） pg/ml, which was significantly higher than that of stimulated with only 10 ng/ml IL-13. MRC-5 in the group of LTD4 and IL-13 stimulated was incubated with Montelukast, the eotaxin was （148.3±15.7） pg/ml, which was significantly lower than that without Montelukast. These results suggested that there be low expression of CysLTIR mRNA in human lung fibralasts. IL-13 can upregulated the expression of CysLTIR in a time- and dose-dependent manner. There is synergistic effect of LTD4 and IL-13 on eotaxin production of fibroblasts, which can be abolished by Montelukast. This effect may be due to upregulatioin of CysLTIR expression

关 键 词：人肺成纤维细胞 白三烯受体1 嗜酸粒细胞趋化因子 白三烯D4 白介素13 MONTELUKAST 

分 类 号：R392[医药卫生—免疫学]