摘要
目的:研究雾化吸入母牛分枝杆菌(微卡)对卵清蛋白(OVA)诱导的哮喘小鼠氧化应激水平的影响及其可能的机制。方法:将18只6~8周Balb/C雄性小鼠随机分为对照组、哮喘组、微卡干预组,每组6只;利用OVA对哮喘组和微卡干预组小鼠致敏和激发建立哮喘模型,微卡干预组在每次激发前予雾化吸入微卡进行干预,对照组予等量生理盐水进行致敏和激发;于末次激发24 h内行肺功能检测;苏木精—伊红(HE)染色、过碘酸雪夫(PAS)染色观察肺组织病理改变、气道杯状细胞增生情况;酶联免疫吸附试验(ELISA)法检测肺泡灌洗液(BALF)中白介素(IL)-4、白介素(IL)-13及干扰素(IFN)-γ水平;检测肺组织丙二醛(MDA)、超氧化物歧化酶(SOD)及还原型谷胱甘肽(GSH)含量;蛋白免疫印迹实验(western blotting)检测核因子E2相关因子(Nrf2)、血红素加氧酶1(HO-1)蛋白表达水平;实时荧光定量聚合酶链反应(RT-qPCR)法检测Nrf2、HO-1 mRNA表达水平。结果:雾化吸入微卡改善哮喘小鼠气道高反应性、肺组织炎症反应及杯状细胞增生(均P<0.05),降低BALF中IL-4和IL-13水平(P<0.01),促进IFN-γ分泌(P<0.01),上调SOD、GSH水平(均P<0.05),减少MDA含量(P<0.01),肺组织Nrf2、HO-1蛋白及mRNA表达增加(均P<0.05)。结论:雾化吸入微卡可能通过调节Nrf2/HO-1通路减轻OVA诱导哮喘小鼠氧化应激。
Objective:To study the effects of aerosol inhalation of Mycobacterium vaccae(M.v.)on oxidative stress in ovalbumin(OVA)-induced asthmatic mice and its possible mechanism.Methods:Eighteen male Balb/C mice of 6-8 weeks were randomly divided into normal control(NC)group,asthma(OVA)group and asthma model withM.v.group,with 6 mice in each group.Mice in OVA group and M.v.group were sensitized and challenged with OVA to establish asthma models.Mice in M.v.group received aerosol inhalation with M.v.before each challenge,while mice in NC group were sensitized and challenged with the same amount of normal saline.Lung function was detected within 24 hours after the last challenge;HE and PAS staining were used to observe the pathological changes of lung tissue and goblet cell proliferation in the airway;the levels of interleukin(IL)-4,interleukin(IL)-13 and interferon(IFN)-γin BALF were detected by ELISA;the contents of malondialdehyde(MDA),superoxide dismutase(SOD)and reduced glutathione(GSH)in lung tissue were measured;the protein expressions of Nrf2 and HO-1 were detected by western blotting and the expression levels of Nrf2 and HO-1 mRNA by real-time fluorescence quantitative polymerase chain reaction(RT-qPCR).Results:Aerosolized inhalation of M.v.improved airway hyperresponsiveness,lung inflammation and goblet cell hyperplasia in asthmatic mice(all P<0.05),decreased the levels of IL-4 and IL-13 in BALF(P<0.01),promoted the secretion of IFN-γ(P<0.01),upregulated the levels of SOD and GSH(all P<0.05),decreased MDA content(P<0.01)as well as increased the mRNA and protein expressions of Nrf2 and HO-1 in lung tissue(P<0.05).Conclusion:Aerosol inhalation of M.v.may reduce the oxidative stress in OVA-induced asthmatic mice by regulating Nrf2/HO-1 pathway.
作者
方莉婷
章谦男
肖欢
李超乾
Fang Liting;Zhang Qiannan;Xiao Huan;Li Chaoqian(Department of Emergency,The First Affiliated Hospital of Guangxi Medical University,Nanning 530021,China)
出处
《广西医科大学学报》
CAS
2022年第12期1931-1936,共6页
Journal of Guangxi Medical University
基金
国家自然科学基金资助项目(No.81860005)
广西自然科学基金资助项目(No.2020GXNSFDA238003,No.2019GXNSFAA185037)。
作者简介
通信作者:李超乾,E-mail:lichaoqiangood@163.com。
