三步序贯法对哮喘模型大鼠激素撤减过程中TGF-β_1/Smad信号通路的影响  

Effect of Three Stage Sequential Therapy on TGF-β_1/Smad Signaling Pathway in Asthma Model Rats in the Process of Steroid Withdrawal

作　　者：田彦 崔红生 李长安 秦芳芳 高誉珊[3] 秦阳 李倩男 

机构地区：[1]北京中医药大学第三附属医院呼吸科,北京100029 [2]北京市通州区新华医院中医科,北京101100 [3]北京中医药大学基础医学院,北京100029

出　　处：《中国中西医结合杂志》2017年第10期1226-1232,共7页Chinese Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金面上项目(No.81273689)

摘　　要：目的观察三步序贯法对哮喘模型大鼠激素撤减过程中TGF-β_1/Smad信号通路上转化生长因子β_1(TGF-β_1)、Smad2、Smad3、Smad6、Smad7蛋白表达的影响。方法将150大鼠随机分为正常对照组、气道重塑组、地塞米松干预组、普米克令舒组及三步序贯组,每组30只。哮喘大鼠激素撤减模型建立方法:以卵蛋白加氢氧化铝致敏,卵蛋白激发,自激发之日开始,于激发前给予地塞米松0.5 mg/kg腹腔注射,连续2周,从第3周开始,按每周0.1 mg/kg速度递减地塞米松用量,至第7周地塞米松全部撤除。三步序贯组在地塞米松干预基础上,于实验第15~28天(撤减前期)给予第一步方颗粒灌胃,第29~63天(撤减中期)给予第二步方颗粒灌胃,第64~77天(撤减后期)给予第三步方颗粒灌胃。气道重塑组仅给予致敏和激发,普米克令舒组在致敏和激发基础上,给予普米克令舒雾化吸入,各组于实验第28、63、77天末次激发2 h后取左肺组织行免疫组化检测,比较各组TGF-β_1、Smad2、Smad3、Smad6、Smad7蛋白表达变化。结果与本组撤减前期比较,地塞米松组撤减中、后期TGF-β_1、Smad2、Smad3蛋白表达升高,Smad6、Smad7蛋白表达降低(P<0.05,P<0.01);普米克令舒组和三步序贯组撤减中、后期TGF-β_1、Smad3、Smad2蛋白表达降低,Smad6蛋白表达升高(P<0.05,P<0.01)。在撤减过程中,与正常对照组同期比较,气道重塑组撤减前、中、后期TGF-β_1、Smad2及Smad3蛋白表达均升高,Smad6、Smad7蛋白表达均降低(P<0.01);与气道重塑组比较,地塞米松组、普米克令舒组、三步序贯组前、中、后期TGF-β_1、Smad2、Smad3蛋白表达均降低,Smad6蛋白表达升高(P<0.01),普米克令舒组、三步序贯组Smad7蛋白表达升高(P<0.01),地塞米松组Smad7蛋白表达撤减前、后期升高(P<0.05);与地塞米松组比较,普米克令舒组、三步序贯组前、中、后期TGF-β_1、Smad2及Smad3均降低,Smad6升高(P<0.01),Smad7中、后期升�Objective To observe the effect of three stage sequential therapy on protein expressions of TGF-β1, Smad2, Smad3, Smad6, Smad7 on TGF-β1/Smad signaling pathway in asthma model rats in the process of steroid withdrawal. Methods Totally 150 SD rats were randomly divided into the normal control group （A）, the airway remodeling group （B）, the Dexamethasone intervening group （C）, the Pulmicort Respules group （D）, and three stage sequential therapy group （E）. The steroid withdrawal model was established in asthma rats： sensitizing with OVA and AI （OH）s, stimulating with OVA, 0.5 mg/kg Dexamethasone was intraperitoneally injected 30 min before every stimulation for 2 successive weeks from starting stimulation. Dexamethasone was reduced at 0.1 mg/kg per week from the 3rd week till completely withdrawing Dexamethasone at week 7. Based on Dexamethasone intervening, rats in Group E were administered by gastrogavage with 1st stage granule from day 15 to day 28 （prophase withdrawal）, 2nd stage granule from day 29 to day 63 （metaphase of withdrawal）, 3rd stage granule from day 64 to day 77 （anaphase withdrawal）. Rats in Group B were only treated with sensitizing and stimulating. Besides sensitizing and stimulating, rats in Group D were aerosol inhaled with Pulmicort Respules. Left lung tissues of every group were sampled for immunohistochemical assay at 2 h after stimulating on day 28, 63, and 77. Protein expressions of TGF-β1, Smad2, Smad3, Smad6, and Smad7 were compared. Results Compared with prophase withdrawal in the same group, protein expressions of TGF- β1, Smad2, and Smad3 increased, protein expressions of Smad6 and Smad7 decreased in Group C in metaphase and anaphase withdrawal （P 〈0.05, P 〈0.01 ） ; protein expressions of TGF-β1, Smad2, and Smad3 decreased, protein expression of Smad6 increased in Group D and E in metaphase and anaphase withdrawal （P 〈0.05, P 〈0.01 ）. In the process of steroid withdrawal, as compared with Group A, protein expressions of TGF-β

关 键 词：三步序贯法 哮喘 激素撤减 转化生长因子Β1 SMADS蛋白 

分 类 号：R-332[医药卫生] R562.25