抗趋化因子配体20抗体对哮喘小鼠气道炎症和气道高反应的抑制作用

发布时间：2018-04-01 03:20

本文选题：哮喘　切入点：单克隆抗体　出处：《第三军医大学学报》2014年12期

【摘要】：目的研究趋化因子配体20(CC chemokine ligand 20,CCL20)在哮喘小鼠肺组织内的表达,观察抗小鼠CCL20单克隆抗体对哮喘小鼠肺部炎症和气道高反应的抑制作用。方法 6~8周龄雌性BALB/c小鼠按随机数字表法分为4组:哮喘组、正常对照组、CCL20单克隆抗体治疗组(单抗组)和治疗对照组,每组8只。哮喘组予卵蛋白(ovalbumin,OVA)致敏和激发建立哮喘小鼠模型,单抗组和治疗对照组致敏和激发同哮喘组,但自第1次激发起,雾化前1 h分别腹腔注射给予抗小鼠CCL20单克隆抗体或磷酸盐缓冲溶液(PBS),连续5 d。正常对照组用PBS代替OVA进行致敏和激发。末次激发后24 h用无创肺功能体描仪行小鼠气道反应性检测,HE染色观察肺组织病理炎症及评分,免疫组化及图像分析和实时荧光定量PCR分别测定小鼠肺组织CCL20的蛋白含量和基因表达水平。结果哮喘组小鼠肺组织炎症病理评分、气道高反应、肺组织CCL20 mRNA相对表达量及免疫组化CCL20平均光密度值均显著高于正常对照组。与哮喘组相比,单抗组小鼠肺部细胞浸润减轻(P0.05),肺功能(雾化乙酰甲胆碱浓度为25~50 mg/mL时)Penh降低(P0.05),肺组织CCL20平均光密度值显著降低(P0.01),而CCL20 mRNA表达无影响。结论 CCL20单克隆抗体可改善OVA诱发的小鼠哮喘炎症和气道高反应,其抑制作用可能与抗鼠CCL20单抗阻断部分CCL20活性有关。
[Abstract]:Objective to investigate the expression of chemokine ligand 20(CC chemokine ligand 20 (CCL20) in lung tissue of asthmatic mice and to observe the inhibitory effect of anti CCL20 monoclonal antibody on pulmonary inflammation and airway hyperresponsiveness in asthmatic mice.Methods female BALB/c mice aged 6 weeks and 8 weeks were randomly divided into four groups: asthma group, normal control group, CCL20 monoclonal antibody treatment group and treatment control group, with 8 mice in each group.Asthma group was sensitized with ovalbumin OVA) and stimulated to establish asthma model. The monoclonal antibody group and treatment control group were sensitized and stimulated with the same asthma group, but since the first stimulation,The mice were injected intraperitoneally with CCL20 monoclonal antibody or phosphate buffer solution 1 h before atomization for 5 days.The normal control group was sensitized and stimulated by PBS instead of OVA.24 hours after the last challenge, the lung tissue inflammation and score were observed by using noninvasive pulmonary function scintigraphy to detect the airway reactivity of mice and to observe the pathological inflammation of lung tissue by HE staining.Immunohistochemistry, image analysis and real-time fluorescence quantitative PCR were used to detect the protein content and gene expression of CCL20 in lung tissue of mice.Results the inflammatory pathological score, airway hyperresponsiveness, the relative expression of CCL20 mRNA and the average optical density of CCL20 in lung tissue of asthmatic mice were significantly higher than those of normal control group.Compared with the asthmatic group, the lung cell infiltration in the monoclonal antibody group decreased P0.05A, and the lung function (the concentration of nebulized methacholine was 25 mg/mL, Penh decreased P0.05N, the mean optical density of CCL20 in lung tissue decreased significantly, but the expression of CCL20 mRNA was not affected.Conclusion CCL20 monoclonal antibody can improve asthmatic inflammation and airway hyperresponsiveness induced by OVA in mice, and its inhibitory effect may be related to the blocking of partial CCL20 activity by anti-mouse CCL20 monoclonal antibody.
【作者单位】：重庆医科大学附属儿童医院儿童发育疾病研究省部共建教育部重点实验室;重庆医科大学附属儿童医院呼吸中心;
【分类号】：R562.25

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