降气平喘方对哮喘大鼠气道平滑肌细胞中ADAM33和ETS-1表达的影响  

Effect of Jiangqi Pingchuan prescription on the expression of a disintegrin and metalloprotease 33 and E26 transformation-specific 1 in airway smooth muscle cells of rats with asthma

作　　者：贺思雨 朱沁泉 张涤[2] HE Siyu;ZHU Qinquan;ZHANG Di(The First Traditional Chinese Medicine Clinical College of Hunan University of Chinese Medicine,Changsha 410007,Hunan,China;The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,Hunan,China)

机构地区：[1]湖南中医药大学第一中医临床学院,湖南长沙410007 [2]湖南中医药大学第一附属医院,湖南长沙410007

出　　处：《湖南中医杂志》2024年第4期141-146,共6页Hunan Journal of Traditional Chinese Medicine

基　　金：湖南省政府特批专项“张涤中医儿科临床研究所专项”(99-16);湖南省教育厅资助项目(18C0397);湖南中医药大学研究生创新课题(2022CX158)。

摘　　要：目的:探讨降气平喘方含药血清对哮喘大鼠气道平滑肌细胞(ASMC)中易感基因解整合素金属蛋白酶33(ADAM33)和E26转录因子-1(ETS-1)表达的影响及其作用机制。方法:通过卵清蛋白(OVA)致敏建立大鼠哮喘模型,另取SD雌性大鼠分组(降气平喘方组、孟鲁司特钠组)灌胃给药制备不同浓度的含药血清,即降气平喘方组血清分别配制为浓度5%、10%、15%的中药含药血清培养液,孟鲁司特钠组血清配制为30%浓度的西药含药血清培养液,分别对体外培养的ASMC进行同化干预24h。采用CCK8法测定不同含药血清对ASMC生长的抑制率,ELISA法检测各组细胞培养上清液中转化生长因子-β1(TGF-β1)、白细胞介素-6(IL-6)的浓度,Western Blot法检测各组细胞中ADAM33、ETS-1蛋白表达情况。结果:培养出的细胞在细胞免疫荧光检测下鉴定成功;5%和10%中药血清组均能明显抑制ASMC的增殖(P<0.05或P<0.01);ASMC培养上清液中,10%中药血清组的TGF-β1和IL-6浓度较哮喘模型组明显降低(P<0.05);与哮喘模型组比较,10%中药血清组对ADAM33蛋白的高表达下调作用更显著(P<0.01),10%、15%中药血清组及30%西药血清组对ETS-1蛋白的高表达下调更显著(P<0.01)。结论:降气平喘方可通过调控ADAM33和转录因子ETS-1表达,减少炎性细胞介质TGF-β1和IL-6释放以及改善气道重塑,达到干预和治疗哮喘的作用。Objective:To investigate the effect of serum containing Jiangqi Pingchuan prescription on the expression of the susceptibility genes a disintegrin and metalloprotease 33(ADAM33)and E26 transformation-specific 1(ETS-1)in airway smooth muscle cells(ASMCs)of rats with asthma and its mechanism of action.Methods:Ovalbumin sensitization was used to establish a rat model of asthma,and female Sprague-Dawley rats were divided into Jiangqi Pingchuan prescription group and montelukast sodium group and were given drug-containing serum at different concentrations.The serum containing 5%,10%,and 15%Jiangqi Pingchuan prescription was prepared for the Jiangqi Pingchuan prescription group,and the serum containing 30%montelukast sodium was prepared for the montelukast sodium group.ASMCs cultured in vitro were assimilated and intervened using the drug-containing serums for 24 hours.CCK8 assay was used to measure the inhibition rate of different drug-containing serums on the growth of ASMCs;ELISA was used to measure the concentration of transforming growth factor-beta 1(TGF-β1)and interleukin-6(IL-6)in the supernatant of each group;Western blot was used to measure the protein expression levels of ADAM33 and ETS-1 in cells.Results:The cells cultured were successfully identified by cellular immunofluorescence technique.The serum containing 5%and 10%Jiangqi Pingchuan prescription significantly inhibited the proliferation of ASMCs(P<0.05 or P<0.01).In the supernatant of ASMCs,the 10%Jiangqi Pingchuan prescription group had significant reductions in the concentration of TGF-β1 and IL-6 compared with the asthma model group(P<0.05).Compared with the asthma model group,the 10%Jiangqi Pingchuan prescription group had significantly greater downregulation of the high expression level of ADAM33 protein(P<0.01),and the 10%and 15%Jiangqi Pingchuan prescription groups and the 30%Western medicine group had significantly greater downregulation of the high expression level of ETS-1 protein(P<0.01).Conclusion:Jiangqi Pingchuan prescription can reduc

关 键 词：哮喘 降气平喘方 气道平滑肌细胞 易感基因 气道重塑 

分 类 号：R285.5[医药卫生—中药学]