健儿乐颗粒对哮喘小鼠Th17/Treg失衡影响的实验研究

Effect of Jian′erle Granule on Th17/Treg Imbalance of Asthma Mice

目的观察支气管哮喘小鼠血清及支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)上清中Th17相关炎症因子IL-17、Treg相关炎症因子IL-10水平,探讨健儿乐颗粒防治哮喘的作用机制。方法40只Balb/c小鼠按体重采用随机数字表法分为正常对照组、哮喘组、激发后哮喘组和健儿乐颗粒加哮喘组,共4组,每组10只。除正常对照组外,各组小鼠采用腹腔注射抗原混悬液后,1%鸡清白蛋白(OVA,gradeⅤ)溶液致敏和激发建立小鼠哮喘模型。造模后健儿乐颗粒加哮喘组小鼠给予健儿乐颗粒(0.36g/mL)药物干预,其余各组给予等量生理盐水灌胃,均持续14天。给药结束后,激发后哮喘组和健儿乐颗粒加哮喘组再次用1%OVA(gradeⅡ)溶液雾化吸入激发约40min一次。HE染色评价小鼠肺部炎症;酶联免疫吸附试验(enzyme linked immunosorbent assay,ELISA)检测血清及BALF上清中IL-10和IL-17水平。结果正常对照组小鼠肺组织肺内支气管结构正常,未见炎症细胞浸润。哮喘组及激发后哮喘组小鼠支气管壁黏膜上皮细胞损伤,有炎症细胞浸润。健儿乐颗粒加哮喘组小鼠支气管和血管周围炎症明显减轻。与正常对照组比较,哮喘组血清及BALF中IL-17水平升高,IL-10水平降低,差异均有统计学意义(P<0.01)。与哮喘组比较,激发后哮喘组血清及BALF中IL-17水平升高,IL-10水平降低,差异均有统计学意义(P<0.01);健儿乐颗粒加哮喘组IL-17水平降低,IL-10水平升高,差异均有统计学意义(P<0.01)。与激发后哮喘组比较,血清及BALF中健儿乐颗粒加哮喘组IL-17水平降低,IL-10水平升高,差异均有统计学意义(P<0.01)。结论健儿乐颗粒防治哮喘的作用机制,可能与其调节小鼠血清、BALF中Th17/Treg细胞因子间的平衡有关。

Objective To observe the levels of Th17 associated inflammatory factor IL-17 and Treg associated inflammatory factor IL-10 in serum and bronchoalveolar lavage fluid（BALF）of bronchial asthma mice,and to explore the mechanism of Jian′erle Granule（JG）for preventing and treating asthma.Methods Totally 40 Balb/c mice were divided into 4 groups according to random digit table,i.e.,the normal control group,the asthma group,the post-excitation asthma group,the JG＋asthma group,10 in each group.Asthma model was established by1%ovalbumin（OVA）（gradeⅤ）solution sensitization and excitation after intraperitoneal injection of antigen suspension in all groups except the normal control group.JG（0.36g/mL）was administrated to mice in the JG＋asthma group after modeling.Equal volume of normal saline was administrated to mice in the rest 3groups.All medication lasted for 14 successive days.After medication mice in the JG＋asthma group and the asthma group were excited with 1% OVA（gradeⅡ）again for 40 min by atomization inhalation.Lung inflammation was examined by HE staining.Levels of IL-10 and IL-17 in BALF and serum were measured by ELISA.Results Bronchial structure in lung tissue was normal in mice of the normal control group,with no inflammatory infiltration seen.Mucosal epithelial cells of bronchial wall were injured in the asthma group and the post-excitation asthma group,with inflammatory infiltration seen.Inflammation around bronchus and blood vessels was obviously attenuated in the JG＋asthma group.Compared with the normal control group,the IL-17 level in serum and BALF significantly increased,IL-10 level significantly decreased in the asthma group（P〈0.01）.Compared with the asthma group,IL-17 level in serum and BALF increased and IL-10 level in serum and BALF decreased in the post-excitation asthma group（P〈0.01）.Compared with the asthma group,the IL-17 level in serum and BALF significantly decreased,IL-10 level significantly increased in the JG＋asthma group（P〈0.01）.Compared with the post-excitation asthma group,the IL-17 level in serum and BALF significantly decreased and IL-10 level in serum and BALF significantly increased in the JG＋asthma group（P〈0.01）.Conclusion The mechanism for JG preventing and treating asthma might be correlated with regulating Th17/Treg cytokine balance in serum and BALF.