陈秋仪 任培中 王佳美 弓雪峰 李芷悦 崔红生

摘要 目的：观察加减乌梅丸颗粒对哮喘大鼠激素干预模型气道重塑及转化生长因子（TGF）-β₁/Smad信号通路的影响。方法：选取30只雄性SD大鼠并随机分为正常组、哮喘组、地塞米松组、布地奈德组及中药组，除正常组外其余组以卵蛋白致敏、雾化激发方法建立哮喘大鼠模型，除正常组和哮喘组外其余均予地塞米松干预并逐步撤减，布地奈德组予以普米克令舒雾化，中药组予以加减乌梅丸颗粒灌胃。光镜下观察各组大鼠肺组织病理形态学改变，测定气道壁面积百分比（Wat%）、气道平滑肌面积百分比（Wam%），免疫组织化学法比较各组Ⅰ型胶原（Collagen Ⅰ）、Ⅲ型胶原（Collagen Ⅲ）表达水平，蛋白质印迹法和荧光定量PCR法比较TGF-β₁、Smad2、Smad3、Smad7蛋白及基因表达水平。结果：与正常组比较，哮喘组大鼠气道壁及平滑肌层明显增厚，管腔变窄，Wat%、Wam%升高，差异有统计学意义（P<0.05），Collagen Ⅰ、Collagen Ⅲ、TGF-β₁、Smad2、Smad3蛋白及基因表达水平均升高，差异有统计学意义（P<0.05），Smad7表达水平降低，差异有统计学意义（P<0.05）。与哮喘组比较，地塞米松组Wat%降低，差异有统计学意义（P<0.05），Collagen Ⅰ、Collagen Ⅲ表达水平降低，差异有统计学意义（P<0.05），TGF-β1、Smad3蛋白及基因表达水平均降低，差异有统计学意义（P<0.05）。与地塞米松组比较，中药组Wat%降低，差异有统计学意义（P<0.05），Collagen Ⅰ、Collagen Ⅲ表达水平降低，差异有统计学意义（P<0.05），TGF-β₁、Smad2、Smad3蛋白及基因表达水平均降低，差异有统计学意义（P<0.05）。结论：加减乌梅丸颗粒可通过调控TGF-β1/Smad信號通路从而减少气道平滑肌增生以及Collagen Ⅰ、Collagen Ⅲ蛋白沉积，抑制气道壁增厚，延缓气道重塑的发展。

关键词  加减乌梅丸颗粒;哮喘大鼠;激素干预;气道重塑;转化生长因子-β₁/Smad信号通路

Effect of ModifiedModified Wumei Pill Granule on Airway Remodeling and TGF-β1/Smad Signal Pathway in Asthma Rat Model with Glucocorticoids Intervention

CHEN Qiuyi，REN Peizhong，WANG Jiamei，GONG Xuefeng，LI Zhiyue，CUI Hongsheng

（Department of Respiratory Medicine，Third Affiliated Hospital，Beijing University of Chinese Medicine，Beijing 100029，China）

Abstract Objective：To observe the effect of Modified Wumei Pill Granule on airway remodeling and TGF-β1/Smad signal pathway in asthma rat model with glucocorticoids intervention.Methods：Totally 30 SD mal rats were randomly divided into the normal group，the asthma group，the dexamethasone group，the budesonide group and the Chinese medicine group.Except for rats in normal group，rat in the other groups were sensitization and stimulation with ovalbumin（OVA）to establish the asthma model.Except for rats in normal and asthma groups，rats in the other groups were treated with dexamethasone and gradually withdrawn.Rats in the budesonide group were inhaled with budesonide.Rats in the Chinese medicine group were poured Modified Wumei Ppill Granule into stomach.Observe the lung histopathological changes under light microscope.Measured the percentage of total wall area（Wat%）and airway smooth muscle area（Wam%）.The expression levels of Collagen I and Collagen III in each group were compared by immunohistochemical method.The expression levels of TGF-β1，Smad2，Smad3，Smad7 protein and gene were compared by Western Blotting and RT-PCR.Results：1）Compared with the normal group，the airway wall and smooth muscle layer of asthmatic rats were significantly thicker，the lumen became narrower，Wat% and Wam% increased（P<0.05），and the expression levels of Collagen I，Collagen III，TGF-β₁，Smad2，Smad3 protein and gene increased（P<0.05），while the expression level of Smad7 decreased（P<0.05）.2）Compared with asthma group，Wat% decreased in dexamethasone group（P<0.05），Collagen I and Collagen III decreased（P<0.05），and TGF-β1，Smad3 protein and gene expression decreased（P<0.05）.3）Compared with dexamethasone group，Wat% decreased（P<0.05），Collagen I and Collagen III decreased（P<0.05），and TGF-beta 1，Smad2，Smad3 protein and gene expression decreased（P<0.05）.Conclusion：By regulating TGF-β₁/Smad signal pathway，Modified Wumei pill granule can reduce the proliferation of airway smooth muscle and the deposition of collagen Ⅰ and collagen Ⅲ protein，inhibit the thickening of airway wall and delay the development of airway remodeling.

Keywords Modified Wumei Pill Granule; Asthmatic rat; Glucocorticoids intervention; Airway remodeling; TGF-β₁/Smad signal pathway

中图分类号：R289.5;R256.12 文献标识码：A doi：10.3969/j.issn.1673-7202.2020.08.005

支气管哮喘是由多种细胞及细胞组分参与的气道慢性炎性疾病，临床以吸入性糖皮质激素（Inhaled Corticosteroid，ICS）为治疗首选药物[1]。然而存在部分哮喘患者对激素治疗不敏感，需长期大剂量使用ICS或全身性糖皮质激素来控制症状，通常将此类哮喘称为激素依赖性哮喘（Steroid-dependent Asthma，SDA）[2]。气道重塑是SDA发生发展的重要因素，可引起气道弹性下降，气流受限持续甚至不可逆，加重哮喘症状。……