哮喘小鼠肺间质巨噬细胞的表型特征分析

Characterization of Pulmonary Interstitial Macrophage Phenotypes in a Mouse Model of Asthma

目的:观察哮喘小鼠肺间质巨噬细胞的表型特征。方法:12只BALB/c小鼠随机分为哮喘组和正常组,每组6只。哮喘组和正常组分别采用卵清白蛋白和PBS致敏和激发。流式细胞仪检测肺间质巨噬细胞和M2型肺间质巨噬细胞的数量;RT-PCR检测肺间质巨噬细胞精氨酸酶1(Arg1)、转谷氨酰胺酶2(TG2)、白细胞介素(IL)-10和诱导型一氧化氮合酶(iNOS)mRNA表达水平;ELISA法检测肺间质巨噬细胞体外培养上清液IL-10和IL-12水平。结果:哮喘组小鼠肺间质巨噬细胞占肺单个核细胞的百分数和M2型肺间质巨噬细胞占肺间质巨噬细胞的百分数明显高于正常组(均P<0.01)。哮喘组小鼠肺间质巨噬细胞Arg1和TG2mRNA表达水平明显高于正常组(均P<0.01),IL-10mRNA表达水平明显低于正常组(P<0.01),iNOS mRNA表达水平与正常组比较差异无统计学意义(P>0.05)。哮喘组小鼠肺间质巨噬细胞体外培养上清液IL-10水平明显低于正常组(P<0.01),IL-12水平与正常组比较差异无统计学意义(P>0.05)。结论:哮喘肺间质巨噬细胞表现为替代活化巨噬细胞的表型特征,并高表达TG2。

Objective： To investigate characterization of pulmonary interstitial macrophage phenotypes in a mouse model of asthma. Methods. Twelve BALB/c mice were randomly divided into two groups, an asthma group （n=6） and a normal control group （n=6）. The murine model of asth- ma was established by sensitization with intraperitoneal injection of ovalbumin and intranasal challenge. The percentages of pulmonary interstitial macrophage and M2 pulmonary interstitial macrophage were detected by flow cytometry. The expression of mRNA of arginase 1 （Arg 1）, transglutaminase 2 （TG2）,IL-10 and inducible nitric oxide synthase （iNOS） in pulmonary inter- stitial macrophage was detected with RT-PCR. The levels of IL-10 and IL-12 in culture superna- tant of pulmonary interstitial macrophage were measured by ELISA. Results. The percentages of pulmonary interstitial macrophage and M2 pulmonary interstitial macrophage in the asthma group were significantly higher than those in the normal control group （all P〈0.01）. The expression of Argl and TG2 mRNA in pulmonary interstitial macrophage in the asthma group was significantly higher than that in the normal control group （P〈0.01） and the expression of IL-10 mR- NA was significantly lower than that in the normal control group （P〈0.01）, but there was no significant difference in the expression of iNOS mRNA in pulmonary interstitial macrophage be- tween the asthma group and the normal control group （P〉0.05）. The level of IL-10 in culture supernatant of pulmonary interstitial macrophage in the asthma group was significantly lower than that in the normal control group （P〈0.01）, but there was no significant difference in the level of IL-12 in culture supernatant of pulmonary interstitial macrophage between the asthma group and the normal control group （P〉0.05）. Conclusion： Pulmonary interstitial macrophages were characterized by alternatively activated macrophage phenotype and high expression of trans- glutaminase 2 in asthma.