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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 observe the expression of bone marrow dendritic cells and the expression of Toll-like receptors (TLR2 and TLR4) in myeloid dendritic cells of asthmatic mice and to explore the effect of warming yang Bushen recipe on the expression of TLR signaling-mediated bone marrow dendritic cells Mechanism of the role. Methods Fifty BALB / c mice were randomly divided into normal control group, model group, traditional Chinese medicine group, western medicine group and integrated traditional Chinese and western medicine group with 10 rats in each group. Each group to make the appropriate intervention. Mouse lung tissue sections were stained with hematoxylin and eosin (HE) to observe the morphological structure and inflammatory infiltration. The levels of IL-4, IL-5 and IFN-γ in BALF were detected by enzyme-linked immunosorbent assay The bone marrow dendritic cells of each group were isolated and cultured. The morphological changes of dendritic cells were observed under inverted phase contrast microscope and scanning electron microscope. The expressions of TLR2 and TLR4 mRNA 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 were thickened with infiltration of inflammatory cells in the surrounding area. The inflammation in each treatment group was relieved to varying degrees. Compared with the normal control group, the expression of IL-4 in alveolar lavage fluid of model group was up-regulated and the level of INF-γ was down-regulated. The difference was statistically significant (P <0.05). Compared with model group, the expression of IL-4 in Chinese medicine group, Western medicine group and Integrative Chinese medicine group was down-regulated, the difference was statistically significant (P <0.05). Compared with the model group, both the Chinese medicine group and the integrated traditional Chinese and Western medicine group could up-regulate the level of INF-γ, the difference was statistically significant (P <0.05). The expression of TLR2 and TLR4 mRNA in bone marrow dendritic cells in model group was significantly higher than that in normal control group (P <0.05), and the expression of TLR2 and TLR4 mRNA in bone marrow dendritic cells were decreased in all treatment groups Significance (P <0.05). Conclusion WeYang Bushen Recipe can reduce airway resistance and reduce airway inflammatory response in asthmatic mice. This may be because Wenshen Fang can significantly up-regulate the secretion of Th1-type cytokines by mediating the signal transduction of BMDC-TLR2 / TLR4 pathway , Down-regulate the expression of Th2-type cytokines, and then regulate the immune balance of Th1 / Th2.