摘要
目的:研究IL-4mRNA和IL-4蛋白在哮喘大鼠CD34+细胞中的转录表达及孟鲁司特(montelukast,MK)对其表达的影响。方法:将SD大鼠随机分为3组:哮喘组、MK组和正常对照组。用卵白蛋白制备大鼠哮喘模型。应用双抗体夹心酶联免疫吸附试验测定血浆中IL-4和γ-干扰素(IFN-γ)浓度;用MiniMACS磁珠分选系统分离骨髓CD34+细胞;采用SYBR GREEN I荧光实时定量PCR法测定CD34+细胞中IL-4 mRNA的相对表达量。采用免疫组化技术测定CD34+细胞中IL-4蛋白的表达量。结果:哮喘组骨髓CD34+细胞中IL-4mRNA和IL-4蛋白的表达量高于其它各组(P<0.01);哮喘组除了IFN-γ水平低外,IL-4浓度和嗜酸性粒细胞(Eos)绝对值都是三组中最高的(P<0.01);除哮喘组外,其余组的各项指标相近(P>0.05)。IL-4 mRNA表达量与IL-4浓度、Eos绝对值呈正相关(P<0.01),与IFN-γ浓度呈负相关(P<0.01)。结论:哮喘大鼠骨髓CD34+细胞中IL-4mRNA的表达增强;孟鲁司特可以下调IL-4mRNA的表达,可能成为其抑制哮喘气道炎症形成的重要机制之一。
Objective ：To investigate the expression of interleukin-4 （IL-4） mRNA and IL-4 protein of CD34＋cells in bone marrow of a rat model of asthma and the modulatory effect of montelukast（MK）. Methods：Male SD rats were randomly divided into three groups： asthma group, MK group and the control group. In the experiment, the rat model of asthma was established by ovalbumin （OVA） sensitization method. The concentrations of IL-4 and interferon-γ （IFN-γ）in plasma were measured by using sandwich ELISA.CD34＋cells were isolated by mini-magnetic activated cell sorting （miniMACS）. The relative content of IL-4 mRNA was detected with SYBR GREEN I fluorescent quantitation PCR method. the expression of IL-4 protein of CD34＋cells in bone marrow was determined with immune-histochemistry techniques. Results：The relatively expressive contents of IL-4 mRNA and IL-4 protein of CD34＋ cells in bone marrow of a rat model of asthma were significantly higher than that of the other groups （P〈0.01）. But the IFN-γ,/level was lower in asthma group, the expressive content of IL-4 protein and the absolute count of Eos were the highest among the three groups （P〈0.01）. Every parameter in the other groups was close to each other except asthma group （P〉0.05）.The expression of IL-4 mRNA correlated positively with that of the concentration of IL-4 and the absolute count of Eos, respectively （P〈0.01）, hut correlated negatively with the concentration of IFN-γ （P〈0.01）. Conclusions：CD34＋ cells strongly expressed IL-4 mRNA in the bone marrow of the rat asthma models.MK significantly depressed the expression of IL-4 mRNA, which may he a key process in modulatory mechanism of asthma.
出处
《金华职业技术学院学报》
2008年第6期37-40,共4页
Journal of Jinhua Polytechnic
基金
2005年金华市科技局资助项目(2005-1-344)
