补肺颗粒对β2受体激动剂耐受型哮喘小鼠PDK1/Akt/PDE4D信号通路及肺功能的影响  

Effect of granules for tonifying lung on PDK1/Akt/PDE4D signaling pathway and lung function inβ2AR agonist-tolerant asthmatic mice

作　　者：周玲羚 严博煜 张莎苒 张笑雪 王虹景 王禹霏 刘伟[2] ZHOU Lingling;YAN Boyu;ZHANG Sharan;ZHANG Xiaoxue;WANG Hongjing;WANG Yufei;LIU Wei(Tianjin University of Traditional Chinese Medicine,Tianjin 301617,China;The Second Affiliated Hospital of Tianjin University of Traditional Chinese Medicine,Tianjin 300250,China)

机构地区：[1]天津中医药大学,天津301617 [2]天津中医药大学第二附属医院,天津300250

出　　处：《现代中西医结合杂志》2024年第24期3378-3385,共8页Modern Journal of Integrated Traditional Chinese and Western Medicine

基　　金：天津市教委科研计划项目(2021ZD018)。

摘　　要：目的探讨补肺颗粒对β2受体激动剂耐受型支气管哮喘(哮喘)小鼠肺功能及3-磷酸肌醇依赖性蛋白激酶1(PDK1)/蛋白激酶B(Akt)/磷酸二酯酶4D(PDE4D)信号通路的影响。方法将64只雌性BALB/C小鼠随机分为空白组、模型组、低沙美特罗组、高沙美特罗组、低补肺颗粒组、高补肺颗粒组、低沙美特罗+低补肺颗粒组、高沙美特罗+高补肺颗粒组,每组8只。空白组不造模,其余组小鼠均采用卵清蛋白(OVA)、脂多糖(LPS)气道滴入致敏结合沙美特罗雾化吸入方法构建β2受体激动剂耐受型哮喘模型。从激发第8周开始,每次激发前30 min,模型组给予生理盐水灌胃;低沙美特罗组和高沙美特罗组给予生理盐水灌胃2 h后,分别雾化吸入0.08 mg/kg、0.16 mg/kg的沙美特罗20 min;低补肺颗粒组和高补肺颗粒组分别给予10 g生药/kg、40 g生药/kg补肺颗粒灌胃;低沙美特罗+低补肺颗粒组给予10 g生药/kg补肺颗粒灌胃2 h后,雾化吸入0.08 mg/kg沙美特罗20 min;高沙美特罗+高补肺颗粒组给予40 g生药/kg补肺颗粒灌胃2 h后,雾化吸入0.16 mg/kg沙美特罗20 min。各组均连续干预2周后,采用动物肺功能分析仪检测小鼠肺功能[用力肺活量(FVC)、第0.1秒用力肺活量(FEV0.1)、FEV0.1/FVC、呼气峰流速(PEF)、最大呼气流量峰值(MMEF)],苏木素-伊红(HE)染色观察小鼠气管及肺组织病理形态,实时荧光定量聚合酶链式反应(Real-time PCR)检测小鼠肺组织中PDK1、Akt、PDE4D、环磷酸腺苷(cAMP)mRNA表达情况。结果模型组FVC、FEV0.1、FEV0.1/FVC、MMEF均明显低于空白组(P均<0.05);与模型组比较,高补肺颗粒组、高沙美特罗+高补肺颗粒组FVC均明显升高(P均<0.05),高沙美特罗组、低补肺颗粒组、低沙美特罗+低补肺颗粒组、高沙美特罗+高补肺颗粒组FEV0.1/FVC均明显升高(P均<0.05),低补肺颗粒组、高补肺颗粒组、高沙美特罗+高补肺颗粒组PEF均明显升高(P均<0.05),补肺颗粒各组Objective It is to investigate the effects of granules for tonifying lung(GTL)on lung function and 3-phosphoinositide-dependent protein kinase 1(PDK1)/protein kinase B(Akt)/phosphodiesterase 4D(PDE4D)signaling pathway inβ2AR agonist-tolerant asthmatic mice.Methods Sixty-four BALB/C female mice were randomly divided into blank group,model group,low-dose salmeterol group,high-dose salmeterol group,low-dose GTL group,high-dose GTL group,low-dose salmeterol+GTL group and high-dose salmeterol+GTL group,with 8 mice in each group.The blank group was not modeled,and the mice of the other groups were sensitized with OVA and LPS by airway drop combined with salmeterol nebulized inhalation to establishβ2AR agonist-tolerant asthmatic models.Starting from the 8th week of sensitization,the model group was given normal saline by gavage at 30 min before each sensitization;the low-dose and high-dose groups of salmeterol were treated with 0.08 mg/kg and 0.16 mg/kg of salmeterol by aerosol inhalation for 20 min,respectively;the low-dose and high-dose groups of GTL were treated with 10 g raw herb/kg and 40 g raw herb/kg of GTL by gavage,respectively;the low-dose salmeterol+GTL group was treated with 0.08 mg/kg of salmeterol by aerosol inhalation in 2 h after gavage with 10 g raw drug/kg of GTL;the high-dose salmeterol+GTL group was treated with 0.16 mg/kg of salmeterol by aerosol inhalation in 2 h after gavage with 40 g raw drug/kg of GTL.After continuously treated for 2 weeks,the lung function(FVC,FEV0.1,FEV0.1/FVC,PEF,MMEF)of mice in each group were detected by animal lung function analyzer,the pathological morphology of trachea and lung tissues of the mice were observed by HE staining,and the mRNA expressions of PDK1,Akt,PDE4D,cAMP in lung tissues of the mice were detected by Real-time PCR.Results The FVC,FEV0.1,FEV0.1/FVC and MMEF in the model group were all significantly lower than those in the blank group(all P<0.05);compared with the model group,FVC was significantly increased in the high-dose GTL group,high-dose salmeterol

关 键 词：支气管哮喘 Β2受体激动剂 补肺颗粒 PDK1/Akt/PDE4D信号通路 环磷酸腺苷 

分 类 号：R-332[医药卫生]