白细胞介素5和嗜酸粒细胞趋化因子在支气管哮喘大鼠肺和骨髓信号传导中的作用  

The roles of interleukin-5 and eotaxin in signal transmission between lung and bone marrow of rat asthmatic models

作　　者：刘春涛[1] 王可[1] 黎联[1] 庞亚敏[1] 

机构地区：[1]四川大学华西医院呼吸科,成都610041

出　　处：《中华结核和呼吸杂志》2006年第8期558-562,共5页Chinese Journal of Tuberculosis and Respiratory Diseases

摘　　要：目的探讨支气管哮喘(简称哮喘)大鼠模型中白细胞介素5(IL-5)和嗜酸粒细胞趋化因子(eotaxin)是否参与肺和骨髓间信号传导,观察不同干预措施的影响。方法清洁级雄性Wistar大鼠40只,按随机数字表法分为正常对照组和哮喘组,每组20只。用卵白蛋白(OVA)致敏和激发制作哮喘模型,于激发后30 min及6、12、24、48 h处死大鼠,分别取外周血涂片、骨髓细胞涂片。肺组织以10％甲醛固定做石蜡切片。将血涂片、骨髓涂片、肺组织切片行苏木精-伊红(HE)染色,观察细胞总数和嗜酸粒细胞(EOS)比例。取不同时间点肺组织匀浆,用酶联免疫吸附测定(ELISA)法测定肺组织匀浆IL-5和eotaxin浓度。将不同时间点的肺组织匀浆与正常对照组和哮喘组大鼠骨髓细胞共同孵育,并分别加入地塞米松(DXM)、半乳凝素3(Galectin-3)、孟鲁司特钠、抗IL-5抗体进行干预,采用抗IL-5、抗IL-SRα、抗eotaxin、抗CD34和抗CC趋化因子受体3(CCR3)多克隆抗体进行免疫组化染色,检测两组骨髓细胞中EOS计数和IL-5+细胞、IL-5Rα+细胞、CCR3+细胞、eotaxin+细胞、CD34+细胞的百分比。结果哮喘大鼠外周血、骨髓、肺组织中EOS数分别为0．020 0±0．002 0、0．023±0．003、0．025 0±0．009 0,正常对照组分别为0．010 0±0．003 0、0．009±0．003、0．009 0±0．002 0,两组比较差异有统计学意义(t值分别为2．547、2．718、2．718,P均<0．05);哮喘大鼠肺组织匀浆于激发6 h的IL-5为(89．3±2．4)pg/ml,12 h的eotaxin水平为(4．9±0．5)pg／ml,48 h均回到基线[(1．45±0．23) pg／ml]水平;除30 min外,各时间点哮喘大鼠肺组织匀浆均能诱导正常对照组或哮喘组大鼠骨髓细胞中CD34+、EOS、IL-5+、IL-SRα+、CCR3+、eotaxin+细胞表达增加,Galectin-3干预哮喘大鼠骨髓细胞后,骨髓EOS、IL-5+、IL-5Rα+、CCR3+、eotaxin+和CD34+细胞表达减少(分别为0．021±0．005、0．074±0．007、0．138±0．014、0．067±0．01Objective To explore the roles of interleukin-5 （IL-5） and eotaxin in signal transmission between lung and bone marrow in rat asthmatic models and to investigate the effects of different interventions on the signal transmission. Methods Forty Wistar rats were randomly assigned to a control group and an asthma group（20 in each group）. The asthmatic model were established by ovalbumin（OVA） sensitizing and challenging. The animals were sacrificed 30 min, 6 h, 12 h , 24 h, and 48 h after challenging, respectively. Slides were prepared from peripheral blood, bone marrow, and lung tissue respectively, and stained with HE. Then the total number and percentage of eosinophils （EOS） were counted. The concentrations of IL-5 and eotaxin in the lung homogenate at the different time points were determined by Enzyme-linked immunosorbent assay（ELISA）. The bone marrow cells of the control rats and sensitized rats were incubated with lung homogenate of the different time points, and treated with dexamethasone （DXM）, Galectin-3, anti-IL-5 polyclonal antibody, and montelukast, respectively. The bone marrow cell slides were stained with anti-IL-5 polyclonal antibody, anti-IL-5Rct polyclonal antibody, anti- eotaxin polyclonal antibody, anti-CD34 polyclonal antibody, and anti-CCR3 polyclonal antibody, respectively. The EOS and immunoactive cells were counted and represented as percentages of the total cells. Result The percentages of EOS in peripheral blood, bone marrow, and lung tissue from the asthma group were 0. 020 0± 0. 002 0,0. 023 ±0. 003, 0. 025 0 ±0. 009 0, and those from the control group were 0. 010 0 ± 0. 003 0,0. 009 ± 0. 003, 0. 009 0 ± 0. 002 0 respectively. The differences were significant between the two groups（ t = 2. 547,2. 718,2.718, all P 〈 0. 05 ）. The peak levels of IL-5 and eotaxin were （ 89.3 ± 2.4 ） pg/ml at 6 h and （4. 9 ±0. 5） pg/ml at 12 h after allergy challenge, respectively, then the levels returned to the baseline value after 48 h （ 1.45 ± 0. 23 ） pg/m

关 键 词：哮喘 白细胞介素5 嗜酸粒细胞趋化因子 大鼠 

分 类 号：R562.25[医药卫生—呼吸系统] R446.5[医药卫生—内科学]