旋毛虫肌幼虫排泄分泌抗原诱导的肠道免疫保护作用研究  

Intestinal immunoprotective effects induced by Trichinella spiralis muscle larval excretory-secretory products

作　　者：那皮沙·居热提 何灵 艾尼瓦尔·吾买尔[1] 白杰[1] NAPISHA Jureti;HE Ling;ANIWA’er Wumai’er;BAI Jie(Xinjiang Medical University College of Pharmacy,Urumqi 830011,China;Chongqing Fengjie Country Hospital,Chongqing 404600,China)

机构地区：[1]新疆医科大学药学院,乌鲁木齐830011 [2]重庆市奉节县医院,重庆404600

出　　处：《中国人兽共患病学报》2023年第7期650-656,共7页Chinese Journal of Zoonoses

基　　金：自治区区域协同创新专项-上海合作组织科技伙伴计划及国际科技合作计划(No.2020E01057)。

摘　　要：目的 研究旋毛虫肌幼虫排泄分泌抗原(Trichinella spiralis muscle larvae excretory-secretory, Ts-Es)诱导小鼠肠道保护性免疫能力以及免疫调节作用。方法 54只BALB/c小鼠随机分为3组,空白对照组、旋毛虫感染组(Ts)、旋毛虫Ts-Es抗原免疫组(Ts-Es)。空白组小鼠腹腔注射PBS;抗原免疫组小鼠腹腔注射0.1 mL的Ts-Es抗原与等量弗氏完全佐剂;旋毛虫感染组腹腔注射PBS和弗氏完全佐剂,每隔7 d注射1次,共3次,末次注射后7 d旋毛虫感染组和Ts-Es抗原免疫组用400条/mL旋毛虫感染性幼虫灌胃攻击感染。解剖取材,检查肠道成虫及肌肉幼虫减虫率,用HE染色法观察肠道病理变化,RT-qPCR法检测小肠中目的基因mRNA表达水平。结果 与旋毛虫感染组相比Ts-Es抗原免疫组成虫和肌幼虫减虫率分别为49%(t=8.109,P<0.05)和67%(t=8.090,P<0.05);与空白对照组相比旋毛虫感染组小肠T-bet(t=5.25,P<0.05)、GATA3(t=2.50,P<0.05)、RORγ-t(t=4.71,P<0.05)、IFN-γ(t=3.17,P<0.05)、IL-4(t=2.60,P<0.05)、IL-5(t=3.05,P<0.05)、IL-17A(t=4.88,P<0.05) mRNA表达量升高,Foxp3表达量没有统计学差异(t=1.55,P>0.05);与旋毛虫感染组相比Ts-Es抗原免疫组T-bet(t=4.23,P<0.05)、RORγ-t(t=4.30,P<0.05)、IFN-γ(t=3.02,P<0.05)、IL-17A(t=3.17,P<0.05) mRNA表达量下降;GATA3(t=3.96,P<0.05)、Foxp3(t=2.76,P<0.05)、IL-4(t=5.16,P<0.05)、IL-5(t=3.69,P<0.05)、IL-10(t=2.61,P<0.05) mRNA表达量升高;与空白对照组相比Ts-Es抗原免疫组GATA3(t=6.4,P<0.05)、Foxp3(t=4.32,P<0.05)、IL-10(t=2.6,P<0.05)、IL-4(t=7.26,P<0.05)、IL-5(t=6.3,P<0.05) mRNA表达量升高均有统计学差异;RORγ-t(t=0.41,P>0.05)、T-bet(t=1.02,P>0.05)、IFN-γ(t=0.09,P>0.05)、IL-17A(t=1.80,P>0.05) mRNA表达量无统计学差异;肠道HE染色结果显示,与旋毛虫感染组相比抗原免疫组炎症好转,肠道HE染色结果显示,与旋毛虫感染组相比抗原免疫组炎症好转。结论 Ts-Es抗原免疫小鼠后可诱发转录因子GATA3,Foxp3相关的混合型免疫应答,This study was aimed at studying the protective immunocompetence and immunoregulatory effects induced by Trichinella spiralis muscle larval excretory-secretory products(Ts-Es)on the intestinesin mice.A total of 54 BALB/c mice were randomly divided into three groups:a control group,T.spiralis infected group,and Ts-Es antigen immunized group.Mice in the control group were intraperitoneally injected with PBS;TS-ES antigen immunized mice received 0.1 mL of TS-ES antigen plus an equal volume of Freund’s complete adjuvant intraperitoneally;the T.spiralis infected group was intraperitoneally injected with PBS and Freund’s complete adjuvant once every 7 days three times;and the T.spiralis infected group and the TS-ES antigen immunized group were intragastrically challenged with 400 strips/mL of T.spiralis infected larvae 7 days after the last injection.We assessed the decreases in intestinal adult worms and muscle larvae.HE staining was used to detect the pathological alterations in the intestinal tract,and RT-qPCR was used to determine the mRNA levels of the target genes in the small intestine.With respect to the T.spiralis infected group,the reduction rates of adult worms and muscle larvae in the Ts-Es antigen immunization group were 49%(t=8.109,P<0.05)and 67%(t=8.090,P<0.05),respectively.T-bet(t=5.25,P<0.05),GATA3(t=2.50,P<0.05),RORγ-t(t=4.71,P<0.05),IFN-γ(t=3.17,P<0.05),IL-4(t=2.60,P<0.05),IL-5(t=3.05,P<0.05),andIL-17A(t=4.88,P<0.05)mRNA expression levels in the small intestine of the T.spiralis infected group were higher than those in the control group,although Foxp3 expression showed no statistically significant difference(t=1.55,P>0.05).The T-bet(t=4.23,P<0.05),RORγ-t(t=4.30,P<0.05),IFN-γ(t=3.02,P<0.05),and IL-17A(t=3.17,P<0.05)mRNA expression in the antigen immunized group was lower than that in the T.spiralis infected group.GATA3(t=3.96,P<0.05),Foxp3(t=2.76,P<0.05),IL-4(t=5.16,P<0.05),IL-5(t=3.69,P<0.05),and IL-10(t=2.61,P<0.05)had higher levels of mRNA expression.The mRNA expression of GATA3(t=6.4,P<0.0

关 键 词：旋毛虫肌幼虫 排泄分泌抗原 肠道免疫 免疫保护作用 

分 类 号：R383.1[医药卫生—医学寄生虫学]