朝医麻黄定喘汤对哮喘小鼠MAPKs/NF-κB/VEGF信号通路的影响

Effects of Korean medicine Mahuang Dingchuan Decoction on MAPKs/NF-κB/VEGF signaling pathway in asthmatic mice

目的:探讨朝医麻黄定喘汤对支气管哮喘小鼠气道炎性反应及对MAPKs/NF-κB/VEGF信号通路表达的影响。方法:清洁级雄性BABL/c小鼠40只,随机分成5组:正常对照组,模型组,麻黄定喘汤低、高剂量组,地塞米松治疗组。用卵清蛋白(OVA)致敏、激发制备小鼠哮喘模型,在末次激发24h后取肺泡灌洗液(BALF)及肺组织。采用免疫吸附法(ELISA)法检测BALF中炎性细胞分类及炎性细胞因子白细胞介素-4(IL-4)、白细胞介素-5(IL-5)、白细胞介素-13(IL-13)的含量;右肺提取蛋白做蛋白印迹(Western Blot)检测肺组织中MAPKs/NF-κB/VEGF通路的表达的变化;左肺组织切片后,进行HE染色、Masson染色及MAPKs/NF-κB/VEGF免疫组织化学染色。结果:与正常对照组比较,模型组小鼠BALF中炎性细胞总数、中性粒细胞、嗜酸性粒细胞、淋巴细胞增多,IL-4、IL-5、IL-13水平显著增高(P<0.05);肺组织MAPKs/NF-κB/VEGF蛋白表达水平显著升高(P<0.01)。与哮喘模型组比较,麻黄定喘汤低、高剂量组和地塞米松治疗组小鼠BALF中炎性细胞总数、中性粒细胞、嗜酸性粒细胞、淋巴细胞、IL-4、IL-5、IL-13水平和肺组织MAPKs/NF-κB/VEGF蛋白表达水平均显著降低(P<0.05)。结论:朝医麻黄定喘汤可抑制哮喘小鼠气道炎性反应的发生,其机制可能是通过部分抑制MAPKs/NF-κB/VEGF信号通路的表达而实现的。

Objective: To investigate the effects of Korean medicine Mahuang Dingchuan Decoction on airway inflammation and expression of MAPKs/NF-κB/VEGF signaling pathway in bronchial asthmatic mice. Methods: Forty male BABL/c mice were randomly divided into 5 groups: normal control group, asthma model group, Mahuang Dingchuan Decoction low dose group, Mahuang Dingchuan Decoction high dose group, dexamethasone treatment group. The model of asthmatic mice was induced by ovalbumin(OVA), and the bronchoalveolar lavage fluid(BALF) and lung tissue were taken after the last excitation for 24 hours. The classification of inflammatory cells and the contents of IL-4, IL-5 and IL-13 in the BALF were detected by ELISA. The changes of the expression of MAPKs/NF-κB/VEGF pathway in right lung tissue were detected by Western blot. Sections of the left lung tissue were stained with HE staining, MASSON trichrome staining and MAPKs/NF-κB/VEGF immunohistochemistry staining. Results: Compared with the normal control group, the levels of inflammatory cell count in BALF IL-4, IL-5 and IL-13 in asthma model group were increased(P<0.05). The expression of MAPKs/NF-κB/VEGF in lung tissue was increased(P<0.01). Compared with asthma model group, the levels of inflammatory cell count in BALF, IL-4, IL-5, IL-13 and the expressions of MAPKs/NF-κB/VEGF were significantly lower in Mahuang Dingchuan Decoction low dose group, Mahuang Dingchuan Decoction high dose group and dexamethasone treatment group(P<0.05). Conclusion: Korean medicine Mahuang Dingchuan Decoction can inhibit the occurrence of airway inflammation in asthmatic mice, and the mechanism may be achieved by partially inhibiting the expression of MAPKs/NF-κB/VEGF signaling pathway.