运脾泻肺化痰汤减轻哮喘大鼠气道黏液高分泌作用机制研究

Relieving effect and mechanism of Yunpi Xiefei Huatan Decoction on airway mucus hypersecretion in asthmatic rats

目的探究运脾泻肺化痰汤由酪氨酸激酶(Lyn)介导并通过磷酸肌醇3激酶(PI3K)/丝氨酸/苏氨酸蛋白激酶(Akt)信号通路对哮喘支气管黏液高分泌的调节作用及其机制。方法将SD大鼠致敏并激发建立哮喘动物模型。按照体重将造模成功大鼠随机分为2组:模型组和中药组,每组各10只;另选正常大鼠10只作为正常组。在造模成功后第16天开始灌胃治疗,正常组、模型组均灌胃0.9%NaCl 2 mL,中药组灌胃运脾泻肺化痰汤10μL·g^-1,均连续4周。进行瑞氏-姬姆萨染色观察大鼠肺泡灌洗液(BALF)中的白细胞(WBC)总数和嗜酸性粒细胞百分比,蛋白质印迹法检测Lyn、PI3K和Akt蛋白表达量(光密度值)。结果正常组、模型组和中药组的WBC总数(×107)分别为6.09±0.33,87.28±18.02和16.68±3.13;这3组的嗜酸性粒细胞百分比(EO%)分别为4.41±0.00,7.96±0.00和5.06±0.00;这3组的Lyn蛋白表达量分别为1.53±0.81,0.26±0.19和1.19±0.57;这3组的p-PI3K蛋白表达量分别为1.25±0.04,7.93±0.89和4.70±1.03;这3组的p-Akt蛋白表达量分别为1.19±0.33,7.30±0.62和3.88±1.28。上述指标:模型组与正常组比较,差异均有统计学意义(均P<0.05);中药组与模型组比较,差异均有统计学意义(均P<0.05)。结论运脾泻肺化痰汤通过Lyn激酶介导PI3K/Akt信号通路下调哮喘炎症因子水平,减轻哮喘气道黏液高分泌。

Objective To investigate the regulation effect and mechanism of Yunpi Xiefei Huatan Decoction(YXHD)on mucus hypersecretion in asthmatic bronchus through phosphoinositol kinase(PI3K)/serine/threonine protein kinase B(Akt)signaling pathway mediates Lyn kinase.Methods SD rats were sensitized and challenged to establish asthma animal model.Successful model rats were divided into two groups according to weight:model group and Chinese traditional medicine(CTM)group with 10 rats in each group;and another 10 normal rats as normal group.On the 16 th day after successful modeling,the rats were given gavage.Rats in the normal group were sensitized by intraperitoneal injection of 2 mL saline;rats in CTM group were sensitized by intraperitoneal injection of YXHD 10μL·g^-1,for 4 weeks.The total number of white blood cells(WBC)and percentage of eosinophils in the rat alveolar lavage fluid(BALF)were observed by Wright-giemsa staining.The expression levels(OD value)of Lyn,PI3K and Akt were detected by Western blot.Results The WBC(×107)in normal group,model group,and TCM group were 6.09±0.33,87.28±18.02 and 16.68±3.13,respectively;the percentages of eosinophil(EO%)in the 3 groups were 4.41±0.00,7.96±0.00 and 5.06±0.00,respectively;the Lyn protein expressions in the 3 groups were 1.53±0.81,0.26±0.19 and 1.19±0.57,respectively;the p-PI3K protein expressions in the 3 groups were 1.25±0.04,7.93±0.89 and 4.70±1.03,respectively;the p-Akt protein expressions in the 3 groups were 1.19±0.33,7.30±0.62 and 3.88±1.28,respectively.Compared between model group and normal group,the difference of the factors were significant(all P<0.05);compared between TCM group and model group,the difference of the factors were significant(all P<0.05).Conclusion YXHD can reduce the levels of inflammatory factors and the mucus hypersecretion of asthma airway through the signal pathway of PI3K/Akt mediated by Lyn kinase.