【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。更多还原

【Abstract】 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),but 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 be a key process in modulatory mechanism of asthma.更多还原