摘要
目的:探讨女贞子多糖(LVFP)与布地奈德联用对哮喘幼鼠免疫调节、气道炎症及氧化应激的影响。方法:从女贞子中提取LVFP,并采用傅里叶变换红外光谱、高压离子色谱及高效凝胶过滤色谱技术对LVFP表征进行鉴定。40只BALB/c幼年小鼠随机分为对照组(Control)、哮喘组(OVA)、OVA+布地奈德组(OVA+budesonide)及OVA+LVFP+布地奈德组(OVA+LVFP+budesonide),每组10只,OVA致敏、激发哮喘症状构建哮喘幼鼠模型,激发周期内OVA+budesonide及OVA+LVFP+budesonide组分别给予布地奈德(3μg/d,吸入)或LVFP[(400 mg/(kg·d),灌胃]+布地奈德(3μg/d,吸入),第28天监测气道反应性,第29天处死幼鼠,ELISA检测支气管肺泡灌洗液(BALF)中炎症因子表达,血细胞计数仪、Wright-Giemsa染色法进行炎症细胞计数,流式细胞术检测BALF中炎症细胞比例变化。Western blot测定肺组织氧化应激及线粒体融合、分裂相关蛋白表达。结果:与OVA组相比,OVA+LVFP+布地奈德组支气管高反应性显著降低(P<0.05),BALF中总炎症细胞数和嗜酸性粒细胞数显著减少(P<0.05),哮喘状态下树突状细胞比例显著降低(P<0.05),而抗炎自然杀伤细胞及调节性T细胞比例升高(P<0.05),炎症因子IL-4、IL-5、IL-9、IL-10、IL-13及TGF-β水平降低(P<0.05)。LVFP与budesonide联用还可抑制氧化相关蛋白硝基络氨酸、超氧化物歧化酶2(SOD2)及线粒体融合和分裂相关蛋白OPA1、线粒体融合蛋白2、动力蛋白相关蛋白1及线粒体分裂蛋白1表达(P<0.05)。结论:LVFP与布地奈德联用可降低支气管高反应性,调节免疫细胞数量,减轻肺部炎症反应,抑制氧化应激,有望用于哮喘治疗。
Objective:To investigate efficacy of Ligustrum vicaryi L.fruit polysaccharide(LVFP)combined with budesonide on immunomodulation,airway inflammation and oxidative stress of asthmatic mice.Methods:LVFP was extracted from Ligustrum vicaryi L.fruit and characterized by Fourier-Transform Infrared,High-Performance Ion Chromatography and High-Performance Gel Filtration Chromatography assay.Forty BALB/c young mice were randomly divided into control group(Control),asthma group(OVA),OVA+budesonide group(OVA+budesonide)and OVA+LVFP+budesonide group(OVA+LVFP+budesonide),with 10 mice in each group.Asthma model was constructed by OVA sensitized and challenged of asthma symptoms.During challenged period,OVA+budesonide and OVA+LVFP+budesonide groups were given budesonide(3 μg/d,inhalation)or LVFP[(400 mg/(kg·d),gavage]and budesonide(3 μg/d,inhalation)respectively.Airway resistance assay was monitored at 28 d.Mice were executed at 29 d,and expressions of inflammatory factors in bronchoalveolar lavage fluid(BALF)were detected by ELISA,number of inflammatory cells was measured by blood cell counter and Wright-Giemsa staining.Percentage changes of inflammatory cells in BALF were detected by flow cytometry,and levels of oxidative stress,mitochondrial fusion and fission related proteins expressions in lung tissues were measured by Western blot.Results:Compared with OVA group,bronchial hyperreactivity was significantly decreased in OVA+LVFP+budesonide group(P<0.05),numbers of total inflammatory cells and eosinophil cells were reduced(P<0.05),dendritic cells were reduced(P<0.05),and proportion of anti-inflammatory natural killer cells and regulatory T cells were increased in BALF under asthmatic status(P<0.05),inflammation factors IL-4,IL-5,IL-9,IL-10,IL-13 and TGF-β levels were decreased(P<0.05).Combination of LVFP and budesonide also inhibited expressions of nitrotyrosine,superoxide dismutase 2(SOD2)and mitochondrial fusion and fission related proteins OPA1,Mitofusin 2,Dynamin-related protein 1 and mitochondrial fission protein 1(
作者
吴振奎
钟丽花
刘延霞
刘玉玲
WU Zhenkui;ZHONG Lihua;LIU Yanxia;LIU Yuling(Department of Pediatrics,Hainan Modern Women’s and Children’s Hospital,Haikou 571100,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2022年第9期1069-1074,1081,共7页
Chinese Journal of Immunology
基金
海南省卫生计生行业科研项目(15A200096)。
关键词
女贞子多糖
布地奈德
联用
哮喘
免疫调节
抗氧化
Ligustrum vicaryiL.fruit polysaccharide
Budesonide
Combination
Asthma
Immunomodulation
Antioxidation
