蝎黄解痉治哮颗粒对支气管哮喘模型小鼠肺组织PI3K/AKT通路的影响

Effect of Xiehuang Jiejing Zhixiao Granules (蝎黄解痉治哮颗粒) on PI3K/AKT Pathway in Lung Tissues of Bronchial Asthma Model Mice

目的探讨蝎黄解痉治哮颗粒治疗支气管哮喘的可能作用机制。方法35只BALB/c小鼠随机分为空白组、模型组、孟鲁司特组和蝎黄解痉治哮低剂量组和高剂量组,每组7只。除空白组外,其余各组采用卵蛋白(OVA)雾化吸入制备哮喘小鼠气道重塑模型。孟鲁司特组及蝎黄解痉治哮低、高剂量组分别按0.5、150、300mg/kg灌胃处理;空白组和模型组以生理盐水200μl/只灌胃,各组均隔日1次。检测各组小鼠支气管肺泡灌洗液(BALF)中的总细胞数和分类炎性细胞数;HE、PAS和Masson染色观察肺组织病理变化;免疫组织化学法检测丝氨酸/苏氨酸激酶(AKT)、缺氧诱导因子1α(HIF-1α)、α平滑肌肌动蛋白(α-SMA)在肺组织中的表达;Western blot法检测肺组织中α-SMA的表达。结果与空白组相比,模型组气道周围有大量炎症细胞浸润,气道杯状细胞、黏液分泌及上皮下纤维化程度明显增加,BALF中细胞总数及嗜酸性粒细胞、中性粒细胞、淋巴细胞数及肺组织AKT、HIF-1α、α-SMA蛋白表达明显升高(P<0.05)。与模型组比较,蝎黄解痉治哮高剂量组和孟鲁司特组炎症细胞浸润明显减少,上皮杯状细胞和黏液减少,气道上皮下纤维化程度降低,细胞总数及各类细胞数明显降低,AKT、HIF-1α、α-SMA蛋白表达明显下降(P<0.05)。结论蝎黄解痉治哮颗粒可改善哮喘模型小鼠气道重塑,以高剂量效果更佳,其机制可能与抑制肺组织PI3K/AKT信号通路及α-SMA、HIF-1α蛋白表达有关。

Objective To explore the possible mechanism of Xiehuang Jiejing Zhixiao Granules(XJZG,蝎黄解痉治哮颗粒)for the treatment of bronchial asthma.Methods Thirty-five female BALB/c mice were randomly divided into blank group,model group,montelukast group,XJZG low-dose and high-dose groups,with 7 rats in each group.The mice model of airway remodeling in asthma was established by aerosol inhalation of ovalbumin(OVA)in all the groups except for blank group.The montelukast group,the XJZG low-dose and high-dose groups were administered with 0.5 mg/kg montelukast,and 150 mg/kg and 300 mg/kg XJZG by gavage,respectively,while the blank group and model group were given normal saline by gavage,with 200μl per mouse;each group received intervention once every other day.The total number of cells and classified inflammatory cells in alveolar lavage fluid(BALF)of mice in each group were recorded,and the pathological changes of lung tissues were identified by HE,PAS and Masson staining.The expression of serine/threonine-protein kinases(AKT),hypoxia inducible factor-1α(HIF-1α)andα-smooth muscle actin(α-SMA)in lung tissue were detected by immunohistochemical method.Western blotting was used to detect the expression ofα-SMA in lung tissue.Results Compared to those of the the blank group,a large number of inflammatory cells infiltrated around the airway in the model group;the number of airway goblet cells and the degree of mucus secretion and subepithelial fibrosis increased significantly;the total number of cells and the number of eosinophils,neutrophils,and lymphocytes,as well as the expression of AKT,HIF-1αandα-SMA protein significantly increased(P<0.05).Compared to model group,high-dose XJZG group and montelukast group significantly reduced the infiltration of inflammatory cells,the number of epithelial goblet cells and mucus,the degree of airway subepithelial fibrosis,the number of total cells and various types of cells,and the expression of AKT,HIF-1αandα-SMA protein(P<0.05).Conclusion XJZG can improve airway remodeling in asthmatic model mice,and high-dose XJZG has better effects.The mechanism may be related to the inhibition of PI3 K/AKT signaling pathway and theα-SMA and HIF-1αproteins expression in the lung tissues.