细芥化痰平喘凝胶贴膏对哮喘小鼠气道炎症的影响

Effect of Xijie Huatan Pingchuan gel plaster on airway inflammation in asthmatic mice

关键词:  细芥化痰平喘凝胶贴膏  穴位敷贴  支气管哮喘  Th1/Th2  气道炎症

Key words:  Xijie Huatan Pingchuan gel plaster  acupoint application  bronchial asthma  Th1/Th2  airway inflammation

摘要:

[目的] 观察细芥化痰平喘凝胶贴膏对哮喘小鼠气道炎症的影响，探讨其治疗支气管哮喘的作用机制。[方法] 选取60只健康雄性BALB/c小鼠，随机分为正常组、哮喘模型组、细芥化痰平喘凝胶贴膏低剂量组、中剂量组、高剂量组、细芥敷贴丸组，每组10只。除正常组外，其余各组采用卵清蛋白（OVA）致敏和激发建立小鼠哮喘模型，取小鼠支气管肺泡灌洗液（BALF）进行炎性细胞的总数及炎细胞分类计数检测，苏木素-伊红（HE）染色、丽春红酸性品红-苯胺蓝（Masson）染色观察肺组织病理学变化。采用酶联免疫吸附法（ELLSA）检测小鼠BALF中γ干扰素（IFN-γ）、白细胞介素-4（IL-4）、白细胞介素-6（IL-6）、白细胞介素-17（IL-17）的含量。[结果] 1）OVA成功致敏和激发，建立小鼠哮喘模型。2）与正常组相比，BALF中炎症细胞总数及以嗜酸性粒细胞（EOS）为主的各分类炎症细胞均显著增加（P＜0.01），细芥化痰平喘凝胶贴膏低中高3个剂量组、细芥敷贴丸组均能不同程度地抑制炎性细胞总数及炎症细胞的增加（P＜0.01），细芥敷贴丸组、细芥化痰平喘凝胶贴膏中剂量组两组对白细胞及EOS等炎症细胞的影响功效相似，差异无统计学意义（P＞0.05）。3）正常组未见肺组织病理改变，哮喘模型组改变最为明显，经细芥化痰平喘凝胶贴膏各剂量组、细芥敷贴丸组干预的各组肺组织病理改变较哮喘模型组减轻。4）与正常组相比，哮喘模型组BALF中的IL-4、IL-6、IL-17含量增多（P＜0.05），IFN-γ水平下降（P＜0.05）。细芥化痰平喘凝胶贴膏中、高剂量组及细芥敷贴丸组治疗后，IL-4、IL-6、IL-17含量均减少，IFN-γ含量增加，差异有统计学意义（P＜0.05）。细芥化痰平喘凝胶贴膏低中高3个剂量组总体呈剂量依赖性。[结论] 细芥化痰平喘凝胶贴膏可降低哮喘小鼠的炎性细胞，减少IL-4、IL-6、IL-17的分泌，增加IFN-γ的分泌，从而改善Th1/Th2失衡状态，可能是细芥化痰平喘凝胶贴膏治疗哮喘的机制之一。

Abstract:

[Objective] To observe the effect of Xijie Huatan Pingchuan gel plaster on airway inflammation in asthmatic mice and explore its mechanism of action in treating bronchial asthma.[Methods] The 60 healthy female BALB/c mice were randomly selected and divided into normal group, asthma model group, low dose group of Xijie Huatan Pingchuan gel plaster, medium dose group, high dose group and Xijie application pill group, with 10 mice in each group. In addition to the normal group, the other groups used ovalbumin (OVA) sensitization and stimulation to establish the mouse asthma model. The total number of inflammatory cells was taken from the mouse bronchoalveolar lavage fluid (BALF) and classified for counting. Hematoxylin eosin (HE) staining and ponceau acid fuchsin aniline blue staining (Masson staining) were used to observe the pathological changes of lung tissue. Detection of BALF in mice by ELISA γ Interferon (IFN-γ), the content of interleukin-4 (IL-4), interleukin-6 (IL-6) and interleukin-17 (IL-17). Enzyme linked immunosorbent assay (ELLSA) was used to detect the content of γ Interferon (IFN-γ), interleukin-4 (IL-4), interleukin-6 (IL-6) and interleukin-17 (IL-17) of BALF in mice.[Results] 1) OVA successfully stimulated the establishment of asthma model in mice. 2)Compared with the normal group, the total number of inflammatory cells in BALF and various categories of inflammatory cells mainly eosinophilia (EOS) were significantly increased (P < 0.01), and the three dose groups of Xijie Huatan Pingchuan gel plaster and Xijie application Pill can inhibit the increase of the total number of inflammatory cells and inflammatory cells in varying degrees (P < 0.01). The effect of Xijie application pill group and the medium dose group of Xijie Huatan Pingchuan gel plaster on leukocytes, EOS and other inflammatory cells was similar, with no significant difference (P > 0.05). 3)No pathological changes of lung tissue were found in the normal group, and the most obvious changes were found in the asthma model group. The pathological changes of lung tissue in each group intervened by Xijie Huatan Pingchuan gel plaster and Xijie application pill group were alleviated compared with the asthma model group. 4)Compared with the normal group, the content of IL-4, IL-6, IL-17 in BALF of asthma model group increased(P < 0.05). The level of IFN-γ decreased(P < 0.05). After the treatment, the contents of IL-4, IL-6, and IL-17 in medium and high dose groups of Xijie Huatan Pingchuan gel plaster and Xijie application pill group decreased, and the content of IFN-γincreased, that the difference was statistically significant (P < 0.05). The effect of resolving asthma relieving was dose-dependent in three dose groups of Xijie Huatan Pingchuan gel plaster.[Conclusion] Xijie Huatan Pingchuan gel plaster can reduce inflammatory cells, reduce the secretion of IL-4, IL-6, IL-17, and increase IFN-γ in asthmatic mice. They can recovery the imbalance of Th1/Th2, which may be one of the mechanisms of Xijie Huatan Pingchuan gel plaster in treating asthma.