血吸虫多肽SJMHE1对哮喘小鼠IL-25、NMU以及ILC2调控作用初步研究  

A preliminary study on the regulation of schistosome polypeptide SJMHE1 on IL-25,NMU and ILC2 in asthmatic mice

作　　者：单文琪 薛菲 常明[1] 汪雪峰[2] SHAN Wen-qi;XUE Fei;CHANG Ming;WANG Xue-feng(Department of Pediatrics,Affiliated Hospital of Jiangsu University,Zhenjiang,Jiangsu 212000;Central Laboratory of Affiliated Hospital of Jiangsu University,Zhenjiang,Jiangsu 212000,China)

机构地区：[1]江苏大学附属医院儿科,江苏镇江212000 [2]江苏大学附属医院中心实验室,江苏镇江212000

出　　处：《热带医学杂志》2021年第9期1103-1108,1133,F0004,共8页Journal of Tropical Medicine

基　　金：国家自然科学基金(81871243);江苏省自然科学基金(BK20141295);江苏省社会发展项目(BE2017697);江苏省社会发展项目(BE2017697);江苏省“六大人才高峰”项目(WSN-009);江苏省“六个一”工程项目(LGY2016055);江苏省血地寄防项目(x201812);江苏大学附属医院科技项目(jdfyRC2015010);镇江市重点社会发展项目(SH2018050)。

摘　　要：目的通过注射卵清蛋白(OVA)建立哮喘小鼠模型并用血吸虫多肽SJMHE1干预,研究SJMHE1对白细胞介素-25(IL-25)、神经介素U(NMU)和2型细胞因子表达水平,以及2型固有淋巴细胞(ILC2)和2型辅助性T细胞(Th2细胞)的调控作用。方法将6~8周BALB/c小鼠随机分为正常组、哮喘组和多肽组,每组6只。正常组小鼠未给予任何干预;哮喘组和多肽组小鼠在第0、7、14天给予腹腔注射抗原混合溶液0.2 m L(含OVA 50μg、10%氢氧化铝,溶于PBS),在第21~28天给予2.5%OVA溶液(溶于PBS)雾化,每次至少40 min,进行哮喘激发;多肽组小鼠第0、7、14天皮下注射多肽乳化液(SJMHE1)100μL;第28天,安乐死法处死所有小鼠。HE染色观察小鼠肺部病理变化,用酶联免疫吸附测定法(ELISA)检测小鼠血清IgE水平,用流式细胞术检测小鼠肺脏ILC2和Th2细胞比例,用荧光定量PCR检测小鼠肺组织IL-25、NMU、IL-4、IL-5、IL-13 mRNA水平。结果哮喘模型成功建立,SJMHE1能够显著减轻哮喘小鼠肺组织炎症反应。哮喘组和多肽组小鼠血清IgE水平较正常组显著增高,差异有统计学意义(P<0.05)。与正常组比较,哮喘组小鼠Th2和ILC2细胞比例显著增高,IL-4、IL-5和IL-13 mRNA相对表达量增高,IL-25和NMU表达量均增高,差异均有统计学意义(P<0.05);与哮喘组比较,多肽组小鼠Th2和ILC2细胞比例显著减低,IL-4、IL-5和IL-13 mRNA相对表达量减低,NMU表达量减低,差异均有统计学意义(P<0.05)。相关性分析显示,IL-25和NMU呈显著正相关(r=0.874,P=0.002)。结论SJMHE1下调NMU的表达量,抑制ILC2和Th2细胞的活化和增殖,减轻哮喘小鼠炎症反应。Objective The asthmatic mouse model was established by ovalbumin(OVA)induction and the schistosome polypeptide SJMHE1 was used to intervene,and the effects of SJMHE1 on interleukin-25(IL-25),neruomedin-U(NMU),type 2 cytokine expression and type 2 innate lymphoid cell(ILC2)and type 2 helper T cells(Th2)were investigated.Methods BALB/c mice at 6 to 8 weeks old were randomly divided into normal group,asthma group and polypeptide group,6 in each group.Normal mice were not given any intervention.Mice in asthma group and polypeptide group were intraperitoneally injected with 0.2 mL antigen mixture solution(containing 50μg OVA,10%aluminum hydroxide,soluble in PBS)on day 0,7 and 14,and atomized with 2.5%OVA solution(soluble in PBS)on days 21 to 28,at least 40 min each time,for asthma stimulation.Mice in polypeptide group were subcutaneously injected 100μL of peptide emulsion(SJMHE1)on day 0,7 and 14.On the 28 th day,all the mice were killed.HE staining was used to observe the pathological changes in the lungs of mice.Serum IgE level was detected by enzyme linked immunosorbent assay(ELISA),the proportion of ILC2 and Th2 cells in the lungs of mice was detected by flow cytometry,and the mRNA levels of IL-25,NMU,IL-4,IL-5 and IL-13 were detected by fluorescence quantitative PCR.Results Asthmatic model was successfully established,and SJMHE1 could significantly reduce the inflammatory response in the lung tissue of asthmatic mice.The serum IgE level in asthma group and polypeptide group was significantly higher than that in normal group,and the difference was statistically significant(P<0.05).Compared with the normal group,the proportions of Th2 and ILC2 cells were significantly increased in the asthmatic group(P<0.05);the mRNA relative expression levels of IL-4,IL-5 and IL-13 were increased(P<0.05);the expression of IL-25 and NMU increased(P<0.05).Compared with the asthma group,the proportions of Th2 and ILC2 cells were significantly decreased in the polypeptide group(P<0.05);the relative expression levels of IL-4,IL-5 and

关 键 词：哮喘 SJMHE1 2型固有淋巴细胞 2型辅助性T细胞 白细胞介素-25 神经介素U 

分 类 号：R562.25[医药卫生—呼吸系统]