定喘汤对中性粒细胞性哮喘小鼠的影响及机制研究  

Effect and Mechanism of Dingchuan Decoction on Neutrophilic Asthma in Mice

作　　者：李三[1,2] 朱黎明 曾丹[2,3] 黄进 戴爱国 LI San;ZHU Liming;ZENG Dan;HUANG Jin;DAI Aiguo(Department of Pulmonary and Critical Care Medicine,Hunan Provincial People’s Hospital,Changsha 410016 Hunan,China;Department of Pulmonary and Critical Care Medicine,the First Affiliated Hospital of Hunan Normal University,Changsha 410016 Hunan,China;Department of Gerontology of Hunan Province,Hunan Provincial People’s Hospital,Changsha 410016 Hunan,China;Hunan University of Chinese Medicine,Changsha 410208 Hunan,China)

机构地区：[1]湖南省人民医院呼吸与危重症医学科,湖南长沙410016 [2]湖南师范大学附属第一医院呼吸与危重症医学科,湖南长沙410016 [3]湖南省人民医院湖南省老年医学研究所,湖南长沙410016 [4]湖南中医药大学,湖南长沙410208

出　　处：《中药新药与临床药理》2020年第12期1428-1434,共7页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金项目(81270093);湖南省中医药科研计划项目(2012-90);长沙市科技计划项目(kq1801088);湖南省科技创新计划项目(2017SK50522)。

摘　　要：目的探讨定喘汤对中性粒细胞性哮喘(Neutrophilic asthma,NA)模型小鼠的影响及机制。方法将42只雌性C57BL/6小鼠随机分为6组,即正常对照组、模型组、地塞米松组(1 mg·kg^-1)及定喘汤低、中、高剂量组(16.5、33、66 g·kg^-1),每组7只。采用鸡卵清白蛋白(OVA)联合弗氏完全佐剂(CFA)致敏和OVA激发建立NA动物模型。OVA激发前灌胃给药(10 mL·kg^-1),每日1次,连续2 d。末次激发24 h后,检测小鼠气道阻力;对外周血及支气管肺泡灌洗液(BALF)中的白细胞分类计数;采用苏木精-伊红(HE)染色观察肺脏组织病理学变化及炎性细胞浸润情况;采用ELISA法检测小鼠BALF上清中的白细胞介素17(IL-17)浓度。结果与正常对照组比较,模型组小鼠的气道阻力明显升高(P<0.01);外周血及BALF中的白细胞总数、中性粒细胞明显升高(P<0.01);肺泡及肺组织结构破坏,支气管及血管周围有大量炎性细胞浸润;BALF中的IL-17水平明显升高(P<0.01)。与模型组比较,定喘汤各剂量组小鼠的气道阻力明显下降(P<0.01);外周血及BALF中的白细胞总数、中性粒细胞数明显减少(P<0.01),气道内炎性细胞浸润减少;BALF中IL-17的水平明显下降(P<0.01)。地塞米松组与模型组之间的差异无统计学意义(P>0.05)。结论定喘汤对NA具有治疗作用,可能与其下调IL-17的表达,减少中性粒细胞浸润,减轻气道炎症及气道高反应有关。Objective To observe the curing effect and mechanism of Dingchuan decoction(DCD)in neutrophilic asthma(NA)mice.Methods Forty-two female C57BL/6 mice were randomly divided into 6 groups(n=7):normal control group,neutrophilic asthma model group,DCD groups low,medium and high dose(16.5,33,66 g·kg^-1)of DCD,and dexamethasone(DEX)treatment group(1 mg·kg^-1).The mice were immunized with chicken egg ovalbumin(OVA)combined with complete Freund’s adjuvant(CFA),and then stimulated with OVA atomization inhalation until appearing asthma symptom to establish the neutrophilic asthma model.Twenty-four hours after the end of stimulation,airway resistance were detected,the number of white blood cells in peripheral blood and bronchial alveolar lavage fiuid(BALF)were counted.Airway inflammatory reaction and inflammatory cell infiltration were detected by hematoxylin and eosin staining(HE),and the expression levels of IL-17 in the bronchial alveolar lavage supernatant was detected by ELISA.Results Compared with the normal control group,in the model group,the airway resistance increased significantly(P<0.01),the white cell and neutrophil in peripheral blood and BALF increased(P<0.01),airway obstruction and a large number of inflammatory cells were seen infiltrated around the peribronchiolar and perivascular areas.the expression of IL-17 in the supernatant of BALF were increased(P<0.01).Compared with the model group,the airway resistance of the DCD treatment groups decreased significantly(P<0.01),the amounts of white cell and neutrophil in peripheral blood and BALF decreased(P<0.01),airway obstruction were relieved and the number of inflammatory cells were decreased around the peribronchiolar and perivascular areas.the expression levels of IL-17 in the supernatant of BALF were decreased(P<0.01).There were no significant differences between the NA group and the DEX group(P>0.05).Conclusion Dingchuan decoction could treat neutrophilic asthma by inducing neutrophil infiltration in the airway,relieving airway inflammation and hyperres

关 键 词：中性粒细胞性哮喘 定喘汤 中性粒细胞 IL-17 炎症 气道高反应 小鼠 

分 类 号：R285.5[医药卫生—中药学]