细粒棘球绦虫微囊对小鼠腹腔巨噬细胞活化及炎症因子水平的影响  

Effects of Echinococcus granulosus microcysts on the activation of peritoneal macrophages and the level of inflammatory factors in mice

作　　者：周璇 侯秋莲[2] 李斌[1] 田凤鸣 蔡璇琳 吕洁 高剑[2,3] 马秀敏 ZHOU Xuan;HOU Qiu-lian;LI Bin;TIAN Feng-ming;CAI Xuan-lin;LV Jie;GAO Jian;MA Xiu-min(State Key Laboratory of Pathogenesis,Prevention,Treatment of Disease Highly Prevalent in Central Asia,Tumor Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830000,China;Department of Human Parasitology of College of Basic Medicine of Xinjiang Medical University;Key Laboraory of Parasite and Vector Biology,Ministry of Health)

机构地区：[1]新疆医科大学附属肿瘤医院,省部共建中亚高发病成因与防治国家重点实验室,新疆乌鲁木齐830000 [2]新疆医科大学基础医学院人体寄生虫学教研室 [3]卫生部寄生虫病原与媒介生物学重点实验室

出　　处：《中国病原生物学杂志》2021年第4期411-418,共8页Journal of Pathogen Biology

基　　金：国家自然科学基金项目(No.82060372);卫生部寄生虫病原与媒介生物学重点实验室开放课题项目(No.WSBKFKT201803);新疆维吾尔自治区研究生科研创新项目(No.XJ2020G196);新疆医科大学临床医学高峰学科校内配套经费资助项目(No.33-0104006020801#)。

摘　　要：目的探索细粒棘球绦虫微囊感染对小鼠腹腔不同表型巨噬细胞活化及腹腔灌洗液中炎症因子水平的影响。方法体外培养细粒棘球绦虫原头蚴形成微囊,通过腹腔注射微囊和原头蚴,分别建立BALB/c小鼠细粒棘球蚴继发感染模型,对照组小鼠腹腔注射等体积的磷酸盐缓冲液(PBS)。于感染后的第8、30、60、90、180、240 d,收集各组小鼠的腹腔灌洗液和腹腔细胞,流式细胞术检测F4/80+CD16/32+(M1型)与F4/80+CD206+(M2型)巨噬细胞比例变化,流式细胞微球阵列法(Cytometric Bead Array,CBA)检测6种炎症因子IL-12p70、TNF-α、IFN-γ、MCP-1、IL-10、IL-6水平。结果感染后240 d,微囊感染组(微囊组)存活囊泡和囊团数分别为(237.67±4.5)、(10.33±1.21)个,与原头蚴感染组(原头蚴组)的囊泡和囊团数(70.33±3.5)、(7.17±1.17)个比较差异有统计学意义(均P<0.01)。微囊组和原头蚴组小鼠腹腔M1型巨噬细胞比例均在感染早期开始升高,8~30 d逐渐升高达峰值,后逐渐下降,在240 d有一定程度回升。感染后30、60、90、180 d,微囊组M1型巨噬细胞比例分别为(78.6±7.52)、(72.95±3.07)、(63.33±3.89)、(28.7±4.84)%,与原头蚴组(66.30±3.47)、(64.03±2.67)、(47.42±2.46)、(18.00±6.77)%比较差异有统计学意义(均P<0.05)。而微囊组和原头蚴组的M2型巨噬细胞细胞比例均在感染早期第8 d处于较高水平,8 d后下降但在感染晚期90~240 d持续升高。感染后8、30、180和240 d,微囊组M2型巨噬细胞比例分别为(60.24±1.35)、(78.6±7.52)、(28.7±4.84)、(51.93±9.49)%,与原头蚴组(56.70±4.25)、(66.30±3.47)、(18.00±6.77)、(43.92±2.66)%比较差异有统计学意义(均P<0.05)。CBA显示,微囊组和原头蚴组腹腔灌洗液中促炎因子IL-12p70和TNF的水平均在8~90 d升高,90 d后下降,其中8、30、60和90 d,微囊组IL-12p70和TNF水平均与原头蚴组比较差异有统计学意义(均P<0.01)。微囊组和原头蚴组促炎因子MCP-1、IL-6均在8~180 d�Objective To explore the effect of Echinococcus granulosus microcyst infection on the activation of peritoneal macrophages and the level of inflammatory factors in the peritoneal lavage fluid of mice.Methods Microcysts of E.granulosus were formed by protoscoleces via in vitro culture.BALB/c murine models of secondary hydatid infection were respectively established via intraperitoneal injection of microcysts and protoscoleces.Mice in the control group were intraperitoneally injected with the same volume of phosphate-buffered saline(PBS).Peritoneal lavage fluid and cells from each group were collected 8,30,60,90,180,and 240 d after infection.Flow cytometry was used to detect the proportion of F4/80+CD16/32+(M1 type)and F4/80+CD206+(M2 type)macrophages.A flow cytometric bead array(CBA)was used to determine levels of six inflammatory cytokines:IL-12 p70,TNF,IFN-γ,MCP-1,IL-10,and IL-6.Results Two hundred and forty d after infection,there were 237.67±4.5 surviving cysts and 10.33±1.21 cystic masses in the group infected with microcysts(the microcyst group).The number of surviving cysts and cystic masses differed significantly from the number of surviving cysts and cystic masses(70.33±3.5 and 7.17±1.17)in the group infected with protoscoleces(the protoscolex group)(P<0.01 for all).In the microcyst and protoscolex groups,the proportion of M1 macrophages in the abdominal cavity was relatively high in the early stage of infection,it increased starting on day 8 and peaked on day 30,it then decreased gradually but rose to an extent on day 240.On days 30,60,90,and 180,the proportion of M1 macrophages in the microcyst group was 78.6±7.52,72.95±3.07,63.33±3.89,and 28.7±4.84%,which differed significantly from the proportion in the protoscolex group(66.30±3.47,64.03±2.67,47.42±2.46,and 18.00±6.77%)(P<0.05 for all).The proportion of M2 macrophages in both the microcyst group and the protoscolex group was high during early infection on day 8 and then decreased,but it continued to increase during the late period of infe

关 键 词：细粒棘球绦虫 微囊 原头蚴 巨噬细胞 炎症因子 

分 类 号：R383.33[医药卫生—医学寄生虫学]