基于Notch/STAT信号通路研究PCG对哮喘EOS的调控作用及分子机制

Molecular mechanism of Pingchuan Granules regulated EOS for asthmatic based on Notch/STAT signaling pathway

目的探讨PCG治疗哮喘的作用靶点及分子作用机制。方法将80只Wistar大鼠随机分为Control组、Model组、平喘颗粒(PCG)组及地塞米松(DXMS)组,每组20只。联合吸入卵白蛋白和氢氧化铝制备哮喘模型大鼠,造模结束前5 d开始灌胃给予相应的药物干预治疗,每组根据成模情况取10只用于后续实验。比较四组支气管肺泡灌洗液(BALF)及血清嗜酸性粒细胞(EOS)计数,EOS阳离子蛋白(ECP)和活化趋化因子(Eotaxin)的表达;采用RT-PCR和Western blot检测肺Notch、STAT信号通路相关蛋白及mRNA的表达。结果Model组BALF及血清EOS计数高于Control组(均P<0.01);PCG组BALF及血清EOS计数低于Model组(均P<0.01)。Model组ECP及Eotaxin含量高于Control组(均P<0.01);PCG组ECP及Eotaxin含量低于Model组(P<0.05或P<0.01)。Model组Notch1、2、4,Hes,Jagged1,Delta4蛋白及mRNA表达高于Control组;Notch3,Delta1、3蛋白及mRNA表达低于Control组(均P<0.01)。PCG组Notch1、2、4,Hes,Jagged1,Delta4蛋白及mRNA表达低于Model组;Notch3,Delta1、3蛋白及mRNA表达高于Model组(P<0.05或P<0.01)。Model组P-STAT1、3、5、6蛋白及mRNA表达高于Control组;P-STAT4蛋白及mRNA表达低于Control组(均P<0.01)。PCG组P-STAT1、3、5、6蛋白及mRNA表达低于Model,P-STAT4蛋白及mRNA表达高于Model组(均P<0.01)。结论PCG通过调控Notch/STAT信号通路降低EOS含量治疗哮喘。

Objective To investigate the therapeutic target and molecular mechanism of Pingchuan Granules in the treatment of asthma.Methods Eighty Wistar rats were randomly divided into Control group,Model group,Pingchuan Granule(PCG)group and Dexamethasone(DXMS)group,with 20 rats in each group.Asthma model rats were prepared by the method of combined inhalation of ovalbumin and aluminum hydroxide.Five days before the end of modeling,the corresponding drug intervention treatment was given by gavage.According to the modeling condition,ten of each group were selected for subsequent experiments.Bronchoalveolar lavage fluid(BALF)and serum eosinophilic granulocyte(EOS)counts,EOS cationic protein(ECP)and activated chemokine expression(Eotaxin)were compared among the four groups.Expression of proteins and mRNA related to Notch and STAT signaling pathways in lung were measured by RT-PCR and Western blot.Results BALF and serum EOS counts in Model group were higher than those in Control group(all P<0.01),BALF and serum EOS counts in PCG group were lower than those in Model group(all P<0.01).The content of ECP and Eotaxin in Model group were higher than those in Control group(all P<0.01),and the content of ECP and eotaxin in PCG were lower than those in Model group(P<0.05 or P<0.01).Notch1,2,4,Hes,Jagged1,Delta4 protein and mRNA expression in Model group were higher than those in Control group,and the expression of Notch3,Delta1,3 protein and mRNA were lower than those in Control group(all P<0.01).Notch1,2,4,Hes,Jagged1,Delta4 protein and mRNA expression in PCG group were lower than those in Model group and the expression of Notch3,Delta1,3 protein and mRNA were higher than those in Model group(P<0.05 or P<0.01).The expression of P-STAT1,3,5,6 protein and mRNA in Model group were higher than those in Control group,and P-STAT4 protein and mRNA were lower than those in Control group(all P<0.01).The expression P-STAT1,3,5,6 protein and mRNA in PCG group were lower than those in Model group,and P-STAT4 protein and mRNA were higher than those in Model group(all P<0.01).Conclusion PCG can reduce EOS by regulating Notch/STAT signaling pathway in the treatment of asthma.