微小膜壳绦虫感染对ICR小鼠小肠LY6A(Sca-1)及IFN-γ、STAT1表达的影响  

Effects of Hymenolepis nana infection on the expression of LY6A(Sca-1),IFN-γand STAT1 in the small intestine in ICR mice

作　　者：赵敬 杨汉蕾 陈素素 牟荣 ZHAO Jing;YANG Han-lei;CHEN Su-su;MOU Rong(Department of Parasitology,School of Basic Medical Sciences,Guizhou Medical University,Guiyang 550025,China;Characteristic and Key Laboratory of Modern Pathogenic Biology,Guizhou Medical University,Guiyang 550025,China)

机构地区：[1]贵州医科大学基础医学院人体寄生虫学教研室,贵阳550025 [2]贵州医科大学现代病原生物学特色重点实验室,贵阳550025

出　　处：《中国人兽共患病学报》2021年第7期611-617,共7页Chinese Journal of Zoonoses

基　　金：贵州省留学人员科技创新项目(黔人项目资助合同[2015]1);贵州省2017大学生创新创业训练计划项目(No.DC201710660035);贵州省普通高等学校青年科技人才成长项目(黔教合KY字[2021]153)联合资助。

摘　　要：目的探讨微小膜壳绦虫感染ICR小鼠小肠组织中LY6A及IFN-γ、STAT1的表达情况。方法采集微小膜壳绦虫成虫标本并收集虫卵制作悬液。将ICR小鼠随机分为对照组和实验组,实验组以定量1 000个/只虫卵灌胃感染,于感染后第2 d和第8 d按照编号处死小鼠获取小肠组织。采用HE染色进行小肠组织病理学观察,RT-PCR技术检测LY6A、IFN-γ和STAT1的mRNA相对表达量,免疫组织化学技术检测小肠中LY6A蛋白阳性细胞的表达,并用PRM技术对LY6A蛋白和STAT1蛋白进行相对丰度定量。结果 HE染色结果显示感染后第8 d在肠腔内发现成虫节片,并且虫体寄生处出现急性炎症反应。RT-PCR检测显示感染第2 d实验组LY6A(t=12.57,P<0.001)和STAT1(t=12.13,P<0.001)的mRNA相对表达量低于对照组,而IFN-γ的mRNA相对表达量高于对照组(t=7.78,P<0.01);感染后第8 d实验组LY6A(t=10.01,P<0.001)和STAT1(t=11.19,P<0.001)的mRNA相对表达量高于对照组;而IFN-γ的mRNA相对表达量低于对照组(t=26.47,P<0.001)。免疫组化结果显示感染后第2 d实验组LY6A阳性细胞百分比高于对照组(t=4.26,P<0.01),感染后第8 d实验组LY6A蛋白阳性细胞百分比高于对照组(t=8.18,P<0.001)。PRM检测结果显示感染后第2 d实验组LY6A蛋白相对表达量低于对照组(t=6.55,P<0.05),实验组STAT1蛋白与对照组相比无统计学差异,感染后第8 d实验组LY6A蛋白(t=4.95,P<0.05)和STAT1(t=2.91,P<0.05)蛋白的相对表达量均高于对照组。结论微小膜壳绦虫感染ICR小鼠小肠后LY6A(Sca-1)的mRNA水平和蛋白水平在幼虫侵入早期呈低表达,成虫期呈高表达;IFN-γ对LY6A的表达不起主导作用,而STAT1可能对LY6A(Sca-1)的表达起诱导作用。This study aimed to investigate the expression of LY6A,IFN-γand STAT1 in the small intestine in ICR mice infected with Hymenolepis nana.Adult specimens of Hymenolepis nana were collected,and the eggs were retrieved and used to make a suspension.ICR mice were randomly divided into a control group and experimental group.The experimental group was infected with 1000 eggs per mouse.On the 2nd and 8th days after infection,the mice were killed according to the number,and small intestinal tissue was obtained.The histopathology of the small intestine was observed with HE staining,the relative expression of LY6A,IFN-γand STAT1 mRNA was detected by RT-PCR,the expression of LY6A protein positive cells in the small intestine was detected by immunohistochemistry,and the relative abundance of LY6A and STAT1 proteins was quantified with the PRM technique.The results of HE staining showed that adult segments were found in the intestinal cavity on the 8th day after infection,and an acute inflammatory reaction was observed in the parasitic part of the parasite.RT-PCR detection showed that the relative expression of LY6A(t=12.57,P<0.001)and STAT1(t=12.13,P<0.001)mRNA on the second day of infection in the experimental group was lower than that in the control group,and the relative expression of IFN-γmRNA in the experimental group was higher than that in the control group(t=7.78,P<0.001).On the 8th day after infection,the relative expression of LY6A(t=10.01,P<0.001)and STAT1(t=11.19,P<0.001)mRNA in the experimental group was higher than that in the control group(t=26.47,P<0.001),and the relative expression of IFN-γmRNA was lower than that in the control group.The immunohistochemistry results showed that the percentage of LY6A positive cells in the experimental group was higher than that in the control group on the second day after infection(t=4.26,P<0.01),and the percentage of LY6A protein positive cells in the experimental group was significantly higher than that in the control group on the 8th day after infection(t=8.18,P<0.001

关 键 词：微小膜壳绦虫 LY6A/Sca-1 ICR小鼠 IFN-Γ STAT1 

分 类 号：R383.3[医药卫生—医学寄生虫学]