[关键词]
[摘要]
【目的】 探讨黄连素抗哮喘机制。【方法】 将32只SD大鼠随机分为正常组、模型组、黄连素组、孟鲁司特组,每组8只。 除正常组,其他各组大鼠采用卵清蛋白(OVA)诱导法构建过敏性哮喘模型。造模成功后,给药7 d。显微镜下对收集的支气 管肺泡灌洗液(BALF)中炎症细胞(包括淋巴细胞、中性粒细胞、巨噬细胞和嗜酸性粒细胞)计数,采用酶联免疫吸附分析 (ELISA)检测血清卵清蛋白特异性免疫球蛋白E(sIgE)和总免疫球蛋白E(IgE)水平,采用ELISA法检测BALF中辅助性T细胞 (Th)1相关细胞因子[干扰素(IFN)-γ、白细胞介素(IL)-2]及Th2相关细胞因子(IL-4、IL-5、IL-10和IL-13)含量,实时定量 聚合酶链反应(RT-qPCR)法检测肺组织中 IFN-γ、IL-2及 IL-4、IL-5、IL-10和 IL-13 mRNA 表达水平,采用苏木素-伊红 (HE)染色法观察肺组织病理形态。【结果】 与正常组比较,模型组 BALF中嗜酸性粒细胞、巨噬细胞、中性粒细胞、淋巴细 胞和总细胞数目升高,血清 sIgE和总 IgE水平升高,Th2相关细胞因子 IL-4、IL-5、IL-10、IL-13在 BALF中的含量及在肺 组织中的mRNA表达水平显著升高,Th1 相关细胞因子 IFN-γ 和 IL-2 在 BALF 中的含量及在肺组织中的 mRNA 表达水平显 著降低,差异均有统计学意义(P < 0.05或 P < 0.01),肺组织 HE 病理染色结果可见支气管和肺泡细胞周围有大量炎症细胞 浸润,平滑肌层和气道壁增厚。与模型组比较,黄连素组及孟鲁司特组上述各指标均得到改善(P < 0.05或 P < 0.01),肺组 织可见炎症细胞浸润减少,气道壁和平滑肌层变薄,且2个治疗组间比较无显著性差异(P > 0.05)。【结论】 黄连素抗哮喘作 用机制与调节Th1/Th2细胞因子失衡有关。
[Key word]
[Abstract]
Objective To observe the anti-asthma mechanism of berberine. Methods Thirty-two SD rats were randomly divided into normal group,model group,berberine group and Montelukast group,eight rats in each group. Except for the normal group, an allergic asthma model was constructed by ovalbumin(OVA)induction method in rats of all other groups. After successful modeling,seven days of drug administration was performed. The inflammatory cells(including lymphocytes,neutrophil,macrophages and eosinophils)were counted in the collected bronchoalveolar lavage fluid(BALF)under microscope, and the serum ovalbumin-specific immunoglobulin E (sIgE)and total immunoglobulin E(IgE)levels were detected by enzyme-linked immunosorbent assay(ELISA), and the levels of T helper cell(Th)1-related cytokines [interferon(IFN)-γ,interleukin(IL)-2] and Th2-related cytokines(IL-4 , IL-5 , IL-10 and IL-13) in BALF were detected by ELISA,and the mRNA expression levels of IFN - γ, IL-2, and IL-4, IL-5, IL-10 and IL-13 in lung tissues were detected by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR),and the histopathologic feature of lung tissues was observed by hematoxylin-eosin (HE)staining. Results Compared with the normal group, the counts of eosinophils, macrophages, neutrophils, lymphocytes and total cells in the model group were increased, the levels of serum sIgE and total IgE were elevated,and the levels of Th2-related cytokines such as IL-4,IL-5, IL-10 and IL-13 in BALF and their mRNA expressions in lung tissue were significantly increased,and the levels of Th1-related cytokines IFN - γ and IL-2 in BALF and their mRNA expression levels in lung tissue were significantly reduced,the difference being statistically significant(P < 0.05 or P < 0.01),and the HE pathological staining results of lung tissue showed that there was a large number of inflammatory cells infiltration around bronchial and alveolar cells,and the smooth muscle layer and the wall of airways were thickened. Compared with the model group,all of the above statistical indexes were improved in the berberine group and the Montelukast group(P < 0.05 or P < 0.01),and the inflammatory cell infiltration was significantly reduced,and airway wall and smooth muscle layer were attenuated in lung tissue, there being no significant differences between the two treatment groups(P > 0.05). Conclusion The mechanism of anti-asthma action of berberine is related to the regulation of Th1/Th2 cytokine imbalance.
[中图分类号]
R285.5
[基金项目]
武汉市卫健委中医药科研项目(编号:WZ22C17)