温阳补肾方对哮喘小鼠骨髓树突细胞TLR信号的调节作用  

Regulating effects of Wenyang Bushen Decoction on TLR signaling in bone marrow dendritic cells of asthmatic mice

作　　者：杨清清 黄伟玲[1] 董志毅[2] 鞠立霞[2] 张素芳[2] 屠洪斌[2] 苏晋梅[2] 李岚[1] 韩明权[2] 王丽新 

机构地区：[1]上海中医药大学附属市中医医院肺病科,上海200071 [2]上海市肺科医院中西医结合科,上海200433

出　　处：《上海中医药杂志》2017年第11期84-90,共7页Shanghai Journal of Traditional Chinese Medicine

基　　金：国家自然科学基金资助项目(81573937)

摘　　要：目的通过观察哮喘小鼠骨髓树突细胞的表达,以及Toll样受体(TLR2和TLR4)在骨髓树突细胞中的表达,探讨温阳补肾方对TLR信号介导的骨髓树突细胞在哮喘发病机制中的作用。方法 50只BALB/c小鼠随机分为正常对照组、模型组、中药组、西药组、中西医结合组,每组10只。各组进行相应干预。小鼠肺组织切片进行苏木素-伊红(HE)染色,观察形态结构及炎症浸润情况;采用酶联免疫吸附法(ELISA)检测肺泡灌洗液中的IL-4、IL-5、IFN-γ的含量;分离培养各组小鼠的骨髓树突细胞,在倒置相差显微镜和扫描电镜下观察树突细胞的形态变化;RT-PCR检测骨髓树突细胞中TLR2和TLR4 mRNA的表达。结果肺组织病理可见模型组小鼠气道黏膜层增厚,周围有明显的炎症细胞浸润,各治疗组炎症均有不同程度的减轻。与正常对照组比较,模型组小鼠肺泡灌洗液中IL-4表达水平上调,INF-γ水平下调,差异有统计学意义(P<0.05)。与模型组比较,中药组、西药组、中西医结合组IL-4表达均下调,差异有统计学意义(P<0.05)。与模型组比较,中药组和中西医结合组均可上调INF-γ水平,差异有统计学意义(P<0.05)。模型组骨髓树突细胞中TLR2、TLR4 mRNA的表达明显高于正常对照组(P<0.05),各治疗组均可下调骨髓树突细胞中TLR2、TLR4mRNA的表达,与模型组比较,差异有统计学意义(P<0.05)。结论温阳补肾方可以降低哮喘小鼠的气道阻力和减轻气道炎性反应,这可能是温肾方通过介导BMDC-TLR2/TLR4通路的信号转导,从而显著上调Th1型细胞因子分泌,下调Th2型细胞因子表达,进而调节Th1/Th2的免疫平衡。Objective To discuss the effects of Wenyang Bushen Decoction on Toll-like receptors（ TLR） signaling mediated bone marrow dendritic cells in the pathogenesis of asthma by observing the expressions of dendritic cells in bone marrow of asthmatic mice and TLR2 and TLR4 expressions in dendritic cells.Methods Fifty BALB/c mice were randomly divided into the normal control group,model group,Chinese medicine group,western medicine group and integrated medicine group,10 mice in each group. Each group was performed the corresponding interventions. The sections of lung tissue were stained with hematoxylin eosin（ HE）,and the morphological structure and inflammatory infiltration were observed. The contents of IL-4,IL-5 and IFN-γ in bronchoalveolar lavage fluid were detected by enzyme linked immunosorbent assay（ ELISA）. The bone marrow dendritic cells of mice in each group were isolated and cultured,and the morphological changes of dendritic cells were observed under inverted phase contrast microscope and scanning electron microscope. The mRNA expressions of TLR2 and TLR4 in bone marrow dendritic cells were detected by RT-PCR. Results The lung tissue pathology showed that the airway mucosa of mice in the model group was thickened and there was obvious inflammatory cell infiltration around it. But the inflammation was alleviated in all treatment groups in different degrees.Compared with the normal control group,the IL-4 content in bronchoalveolar lavage fluid was increased and the INF-γ content was decreased in the model group,with statistically significant differences（ P 0. 05）. Compared with the model group,the IL-4 content was decreased in the Chinese medicine group,western medicine group and integrated medicine group,with statistically significant differences（ P 0. 05）; and the INF-γ content was increased in the Chinese medicine group and integrated medicine group,with statistically significant differences（ P 0. 05）. The mRNA expressions of TLR2 and TLR4 in bone marrow dendritic cells in the model

关 键 词：支气管哮喘 骨髓树突细胞 温阳补肾方 

分 类 号：R285.5[医药卫生—中药学]