巴西日圆线虫缓解葡聚糖硫酸钠盐诱导小鼠溃疡性结肠炎的初步研究  

Nippostrongylus brasiliensis alleviates dextran sulfate sodium salt-induced ulcerative colitis in mice:a preliminary study

作　　者：张莹舒 袁彩怡 王强 丁昕 姚甲凯 张蓓 乔树苗 戴洋 ZHANG Yingshu;YUAN Caiyi;WANG Qiang;DING Xin;YAO Jiakai;ZHANG Bei;QIAO Shumiao;DAI Yang(School of Public Health,Nanjing Medical University,Nanjing,Jiangsu 211166,China;National Health Commission Key Laboratory of Parasitic Disease Control and Prevention,Jiangsu Provincial Key Laboratory on Parasite and Vector Control Technology,Jiangsu Provincial Medical Key Laboratory,Jiangsu Institute of Parasitic Diseases,Wuxi,Jiangsu 214064,China)

机构地区：[1]南京医科大学公共卫生学院,江苏南京211166 [2]国家卫生健康委员会寄生虫病预防与控制技术重点实验室、江苏省寄生虫与媒介控制技术重点实验室、江苏省医学重点实验室、江苏省血吸虫病防治研究所,江苏无锡214064

出　　处：《中国血吸虫病防治杂志》2024年第5期450-459,473,共11页Chinese Journal of Schistosomiasis Control

基　　金：江苏省科教能力提升工程项目(ZDXYS202207);江苏省无锡市卫生健康委员会面上项目(m202221)。

摘　　要：目的观察巴西日圆线虫(Nipponstrongylus brasiliensis,Nb)感染对葡聚糖硫酸钠盐(dextran sulphate sodium salt,DSS)诱导的小鼠溃疡性结肠炎的缓解作用,并初步探讨其作用机制。方法将30只SPF级雄性C57BL/6J小鼠(体质量约25 g)随机分为3组,分别为空白对照组(NC组)、DSS造模组(DSS组)以及巴西日圆线虫干预组(Nb+DSS组),每组10只。DSS组小鼠于实验第1天(D0)开始每日口服3.5%DSS,至D6换为正常饮用水;在DSS组小鼠口服3.5%DSS 5 d前,Nb+DSS组小鼠按500条/鼠的剂量经皮下注射NbⅢ期幼虫进行感染,并于D0开始每日口服3.5%DSS溶液,至D6换为正常饮用水;NC组小鼠给予正常饮用水。实验期间,观察各组小鼠体质量及粪便性状,并进行疾病活动指数(disease activity index,DAI)评分。各组小鼠均于D9剖杀,取结肠测量长度;收集小鼠结肠组织样本,采用苏木精-伊红染色观察组织病理学变化,进行病理学评分。采用实时荧光定量PCR(quantitative fluorescent real-time PCR,qPCR)法及酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测小鼠结肠组织白细胞介素(interleukin,IL)-1β、IL-10 mRNA和蛋白表达水平;采用qPCR和免疫荧光法检测小鼠结肠黏膜修复相关分子闭锁小带蛋白1(zonula occludens-1,ZO-1)、黏蛋白2(mucin-2,MUC2)、封闭蛋白1(claudin-1)m RNA和蛋白表达水平。结果D9时,NC组、DSS组、Nb+DSS组小鼠体质量分别为(26.26±1.93)、(22.39±1.65)、(25.00±1.58)g(F=8.06,P<0.01),3组小鼠DAI评分分别为(1.89±0.34)、(0.47±0.39)、0分(F=57.61,P<0.0001),结肠长度分别为(42.50±5.75)、(56.20±5.96)、(61.17±7.88)mm(F=13.72,P<0.001);两两比较结果显示,D9时,Nb+DSS组小鼠体质量较DSS组显著增加(P<0.05)、DAI评分显著降低(P<0.0001)、结肠长度显著增加(P<0.01)。病理学观察结果显示,与DSS组相比,Nb+DSS组小鼠结肠隐窝相对完整、炎性细胞浸润水平较低。NC组、DSS组、Nb+DSS组小鼠结肠组织病理学评分分别为Objective To investigate the alleviation of Nippostrongylus brasiliensis infection on dextran sulfate sodium salt(DSS)-induced ulcerative colitis in mice,and to explore the underlying mechanism.Methods Thirty male C57BL/6J mice of the SPF grade,each weighing approximately 25 g,were randomly divided into three groups,including the blank control group(NC group),DSS modeling group(DSS group),and N.brasiliensis treatment group(Nb+DSS group),of 10 mice in each group.Mice in the DSS group were orally administered with 3.5%DSS daily since day 1(D0)for 6 successive days,and given normal drinking water since D6,and animals in the Nb+DSS group were subcutaneously injected with the third-stage larvae of N.brasiliensis at a dose of 500 larvae per mice 5 days prior to D0,followed by oral administration with 3.5%DSS daily since D0 for 6 successive days and normal drinking water since D6,while mice in the NC group were given normal drinking water.Mouse body weight and stool were observed and the disease activity index(DAI)was scored in each group during the study period.All mice were sacrificed on D9.The mouse colon length was measured,and mouse colon specimens were subjected to hematoxylin-eosin(HE)staining and histopathological scoring.In addition,the mRNA and protein expression of interleukin(IL)-1βand IL-10 was quantified in mouse colon specimens using quantitative fluorescent real-time PCR(qPCR)assay and enzyme-linked immunosorbent assay(ELISA),and the mRNA and protein expression of mucosal repair-associated molecules zonula occludens-1(ZO-1),mucin 2(MUC2)and claudin-1 was detected in mouse colon specimens using qPCR assay and immunofluorescence assay.Results The mice body weights,DAI scores and colon lengths were(26.26±1.93),(22.39±1.65),(25.00±1.58)g(F=8.06,P<0.01);(1.89±0.34),(0.47±0.39),0 points(F=57.61,P<0.0001);and(42.50±5.75),(56.20±5.96)mm and(61.17±7.88)mm(F=13.72,P<0.001)in the NC,DSS and Nb+DSS groups on D9,respectively,and elevated mouse body weight(P<0.05),reduced DAI score(P<0.0001)and increased colo

关 键 词：巴西日圆线虫 抗炎作用 黏膜修复 溃疡性结肠炎 炎症性肠病 小鼠 

分 类 号：R383.19[医药卫生—医学寄生虫学]