摘要
目的研究达比加群酯改善支气管哮喘小鼠模型气道重塑的机制。方法将48只Wistar小鼠根据干预方式分为正常对照组、哮喘组、凝血酶组和达比加群酯组,按照激发浓度将哮喘组小鼠分为激发1组和激发2组。建立支气管哮喘小鼠模型,取小鼠肺支气管肺泡灌洗液(BALF)、血清及肺组织。采用苏木精-伊红染色(HE)法观察各组小鼠肺组织形态,测量支气管管壁及平滑肌厚度,采用瑞式-姬姆萨染色法观察BALF和血清各类型白细胞计数,采用双抗体夹心法测定BALF、血清中白细胞介素-6(IL-6)和白细胞介素-17(IL-17)水平,采用免疫组化染色法测定肺组织中Yes相关蛋白(YAP)表达量,利用逆转录-聚合酶链反应测定肺组织中YAP mRNA的表达量。结果与正常对照组比较,激发1组、激发2组、凝血酶组和达比加群酯组小鼠支气管管壁及平滑肌厚度增加,BALF和血清中嗜酸性粒细胞、淋巴细胞、单核细胞及中性粒细胞比例、IL-6及IL-17水平均升高,肺组织中YAP蛋白及YAP mRNA表达量均升高,组间比较差异有统计学意义(P<0.05);与激发1组、激发2组和凝血酶组比较,达比加群酯组小鼠支气管管壁及平滑肌厚度减小,BALF和血清中嗜酸性粒细胞、淋巴细胞、单核细胞及中性粒细胞比例降低,肺组织中YAP蛋白及YAP mRNA表达量降低(P<0.05)。结论达比加群酯可能通过降低YAP表达量抑制Hippo信号通路,缓解支气管哮喘小鼠血液高凝、减轻气道炎性反应、抑制气道重塑。
Objective To investigate the mechanism of dabigatran etexilate improving airway remodeling in bronchial asthma mice model.Methods 48 Wistar mice were divided into normal control group,asthma group,thrombin group and dabigatan etexilate group according to different intervention methods.Mice in asthma group were divided into motivate 1 group and motivate 2 group according to different stimulation concentration.After the model establishment,the bronchoalveolar lavage fluid(BALF),serum and lung tissue were taken from mice.The morphology of lung tissue was observed by hematoxylin-eosin(HE)staining.The thickness of bronchial wall and smooth muscle was measured.The white blood cell count in BALF and serum was observed by Wright-Giemsa staining.The levels of BALF and serum interleukin-6(IL-6)and interleukin-17(IL-17)were measured by double antibody sandwich method.The level of Yes-associated protein(YAP)in lung tissue was detected by immune histochemical staining.The relative expression level of YAP mRNA in lung tissue was determined by reverse transcription-polymerase chain reaction.Results Compared with the normal control group,there was an increase in bronchial wall and smooth muscle thickness,proportion of eosinophils,lymphocytes,monocytes and neutrophils in BALF and serum,as well as the expression level of YAP protein and YAP mRNA in lung tissue of motivate 1 group,motivate 2 group,thrombin group and dabigatan etexilate group,with statistic difference(P<0.05).Compared with motivate 1 group,motivate 2 group and thrombin group,there was a decrease in the bronchial wall and smooth muscle thickness,proportion of eosinophils,lymphocytes,monocytes and neutrophils in BALF and serum,as well as the expression level of YAP protein and YAP mRNA in lung tissue in dabigatan etexilate group,with statistic difference(P<0.05).Conclusion Dabigatran etexilate reduces the level of YAP,inhibits the Hippo signal pathway,alleviates the hypercoagulability of blood,reduces the airway inflammation,thus inhibits the airway remodeling in asthmatic mice.
作者
杨丰鹤
高辉
高炜
阎玥
YANG Fenghe;GAO Hui;GAO Wei;YAN Yue(Department of Respiratory Medicine,Renhe Hospital,Beijing 102600,China;Beijing University of Traditional Chinese Medicine,Beijing 100029,China)
出处
《西北药学杂志》
CAS
2020年第5期689-695,共7页
Northwest Pharmaceutical Journal
基金
北京市自然科学基金重点项目(编号:7121013)。
