不同剂量卵蛋白诱发BALB/c小鼠支气管哮喘模型的比较
|
摘要
目的探讨不同剂量卵蛋白(ovalbumin,OVA)对BALB/c小鼠支气管哮喘模型血清Ig E、IL-4、IFN-γ含量、支气管肺泡灌洗液(bronchoalveolar lavage fluid,BALF)嗜酸性粒细胞(eosinophils,EOS)数量及肺组织病理学的影响,初步确定较为合适的OVA造模剂量。方法 BALB/c小鼠随机分为正常组,模型1组(OVA 20μg),模型2组(OVA 50μg),模型3组(OVA 100μg),醋酸地塞米松组(OVA 100μg)。除正常组外,其余各组小鼠分别腹腔注射相应剂量OVA致敏,并以1%OVA雾化激发,建立支气管哮喘模型。ELISA法检测血清Ig E、IL-4、IFN-γ含量,瑞氏染色计算BALF中EOS数目,HE染色观察肺组织病理学变化。结果与正常组相比,各组小鼠血清Ig E、IL-4、IFN-γ及BALF中EOS均显著增加(P<0.05),肺组织中均出现炎性细胞浸润、支气管上皮细胞变性、支气管平滑肌细胞增生等病理改变,在Ig E、IL-4、IFN-γ含量及肺组织病理变化方面以OVA 50μg组小鼠更为明显。结论三种剂量OVA均可成功诱发BALB/c小鼠出现哮喘,综合而言50μg OVA所诱发的哮喘模型更为明显,推荐作为BALB/c小鼠哮喘模型的适宜造模剂量。
Objective To investigate the effects of different doses of ovalbumin(OVA) on the serum levels of Ig E,IL-4,and IFN-γ,on the number of eosinophils(EOS) in bronchoalveolar lavage fluid(BALF),and on lung histopathology in the BALB/c mice with bronchial asthma,to determine the optimal OVA dose.Methods BALB/c mice were randomly divided into a normal group,model 1 group(OVA 20 μg),model 2 group(OVA 50 μg),model 3 group(OVA 100 μg),and dexamethasone acetate group(OVA 100 μg).In addition to the normal group,the other groups of mice were sensitized by intraperitoneal injection of the corresponding doses of OVA,and stimulated by 1% OVA atomization to establish bronchial asthma models.The serum levels of Ig E,IL-4,and IFN-γ were determined by ELISA.The number of EOS in BALF was calculated by Wright staining,and the pathological changes of lung tissue were observed by HE staining.Results Compared with the normal group,the serum levels of Ig E,IL-4,and IFN-γ and EOS in BALF were significantly increased in all groups(P<0.05).Inflammatory cell infiltration,bronchial epithelial cell degeneration,bronchial smooth muscle cell hyperplasia,and other pathological changes were observed in the lung tissues.Especially,in the OVA 50 μg group,the changes in the Ig E,IL-4,and IFN-γ levels and lung histopathological changes were more pronounced.Conclusions All the three doses of OVA can induce asthma in BALB/c mice successfully,and the asthma model induced by 50 μg of OVA had more pronounced alterations.It is recommended that 50 μg of OVA be used as the appropriate dosage for establishing an asthma model in BALB/c mice.
Objective To investigate the effects of different doses of ovalbumin(OVA) on the serum levels of Ig E,IL-4,and IFN-γ,on the number of eosinophils(EOS) in bronchoalveolar lavage fluid(BALF),and on lung histopathology in the BALB/c mice with bronchial asthma,to determine the optimal OVA dose.Methods BALB/c mice were randomly divided into a normal group,model 1 group(OVA 20 μg),model 2 group(OVA 50 μg),model 3 group(OVA 100 μg),and dexamethasone acetate group(OVA 100 μg).In addition to the normal group,the other groups of mice were sensitized by intraperitoneal injection of the corresponding doses of OVA,and stimulated by 1% OVA atomization to establish bronchial asthma models.The serum levels of Ig E,IL-4,and IFN-γ were determined by ELISA.The number of EOS in BALF was calculated by Wright staining,and the pathological changes of lung tissue were observed by HE staining.Results Compared with the normal group,the serum levels of Ig E,IL-4,and IFN-γ and EOS in BALF were significantly increased in all groups(P<0.05).Inflammatory cell infiltration,bronchial epithelial cell degeneration,bronchial smooth muscle cell hyperplasia,and other pathological changes were observed in the lung tissues.Especially,in the OVA 50 μg group,the changes in the Ig E,IL-4,and IFN-γ levels and lung histopathological changes were more pronounced.Conclusions All the three doses of OVA can induce asthma in BALB/c mice successfully,and the asthma model induced by 50 μg of OVA had more pronounced alterations.It is recommended that 50 μg of OVA be used as the appropriate dosage for establishing an asthma model in BALB/c mice.
引文
[1]吴佳佳,钟大玲,刘茈蕊,等.大黄素对哮喘小鼠肺组织形态及BALF中炎性细胞因子分泌水平的影响[J].四川中医,2017,35(04):66-68.
[2]张明强.IRAK-M在哮喘中的免疫调节作用及变应性支气管肺曲菌病患者临床特征分析[D].北京:北京协和医学院,2016.
[3]程胜,陈辉龙,王正云,等.滴鼻和雾化两种不同激发方式对小鼠支气管哮喘模型气道炎症的影响[J].华中科技大学学报(医学版),2014,43(2):121-124.
[4]房祥峰.气管内注射TSLP中和抗体抑制哮喘小鼠气道炎症和气道高反应性研究[D].广州:南方医科大学,2014.
[5]刘家齐,赵正晓,魏颖,等.芍药苷对哮喘模型小鼠气道炎症趋化因子及受体的干预作用[J].中国实验动物学报,2016,24(5):460-464.
[6]刘瑞.厚朴麻黄汤对哮喘小鼠肺组织病理及血清IL-13、INF-γ影响的实验研究[D].郑州:河南中医药大学,2016.
[7]谢林艳.粉防已碱下调NF-κB和i NOS、抑制哮喘小鼠气道炎症和气道高反应性的研究[D].南京:南京医科大学,2015.
[8]王利玲,吴强鹏,程黎.抗菌肽LL-37激活嗜酸性粒细胞释放炎性递质对哮喘的发病机制研究[J].医学研究生学报,2017,30(1):70-76.
[9]李航,刘小虹,徐卫方.中药对哮喘T细胞亚型免疫调节作用的研究进展[J].中药新药与临床药理,2017,28(1):132-136.
[10]龚臣,邓静敏.Th17/Treg在支气管哮喘发病机制中的作用及研究进展[J].中华哮喘杂志(电子版),2013,7(3):41-45.
[11]赵欣欣,万力生,李佳曦,等.基于TGF-β1/Smads信号通路中医药防治哮喘气道重塑研究进展[J].辽宁中医药大学学报,2016,18(1):160-163.
[12]杨今实,金香,孙天一,等.朝医麻黄定喘汤对哮喘小鼠MAPKs/NF-κB/VEGF信号通路的影响[J].中华中医药杂志,2017,32(10):4448-4452.
[13]郑凌霄,于芬芳,刘曼曼,等.哮喘小鼠动物模型的建立与评价[J].热带病与寄生虫学,2017,15(4):244-247.
[2]张明强.IRAK-M在哮喘中的免疫调节作用及变应性支气管肺曲菌病患者临床特征分析[D].北京:北京协和医学院,2016.
[3]程胜,陈辉龙,王正云,等.滴鼻和雾化两种不同激发方式对小鼠支气管哮喘模型气道炎症的影响[J].华中科技大学学报(医学版),2014,43(2):121-124.
[4]房祥峰.气管内注射TSLP中和抗体抑制哮喘小鼠气道炎症和气道高反应性研究[D].广州:南方医科大学,2014.
[5]刘家齐,赵正晓,魏颖,等.芍药苷对哮喘模型小鼠气道炎症趋化因子及受体的干预作用[J].中国实验动物学报,2016,24(5):460-464.
[6]刘瑞.厚朴麻黄汤对哮喘小鼠肺组织病理及血清IL-13、INF-γ影响的实验研究[D].郑州:河南中医药大学,2016.
[7]谢林艳.粉防已碱下调NF-κB和i NOS、抑制哮喘小鼠气道炎症和气道高反应性的研究[D].南京:南京医科大学,2015.
[8]王利玲,吴强鹏,程黎.抗菌肽LL-37激活嗜酸性粒细胞释放炎性递质对哮喘的发病机制研究[J].医学研究生学报,2017,30(1):70-76.
[9]李航,刘小虹,徐卫方.中药对哮喘T细胞亚型免疫调节作用的研究进展[J].中药新药与临床药理,2017,28(1):132-136.
[10]龚臣,邓静敏.Th17/Treg在支气管哮喘发病机制中的作用及研究进展[J].中华哮喘杂志(电子版),2013,7(3):41-45.
[11]赵欣欣,万力生,李佳曦,等.基于TGF-β1/Smads信号通路中医药防治哮喘气道重塑研究进展[J].辽宁中医药大学学报,2016,18(1):160-163.
[12]杨今实,金香,孙天一,等.朝医麻黄定喘汤对哮喘小鼠MAPKs/NF-κB/VEGF信号通路的影响[J].中华中医药杂志,2017,32(10):4448-4452.
[13]郑凌霄,于芬芳,刘曼曼,等.哮喘小鼠动物模型的建立与评价[J].热带病与寄生虫学,2017,15(4):244-247.