包虫囊液干预下宿主限制性因子IFITM3和Foxp3之间的关联研究  

Association study between the host limiting factor IFITM3 and Foxp3 under the intervention of hydatid cyst fluid

作　　者：杨雨阳 祁晶晶[4] 吕洁 齐新伟[2] 马秀敏[2,5] 单骄宇[1,2,3] YANG Yu-yang;QI Jing-jing;LV Jie;QI Xin-wei;MA Xiu-min;SHAN Jiao-yu(Department of Human Parasitology,Basic Medicine College,Xinjiang Medical University,Urumqi 830017,China;Xinjiang Laboratory of Hydatid Functional Medicine,the First Affiliated Hospital;Xinjiang Key Laboratory of Molecular Biology of Endemic Diseases;Inspection of the Fifth Affiliated Hospital;Laboratory Department of the Third Affiliated Hospital)

机构地区：[1]新疆医科大学基础医学院人体寄生虫学教研室,新疆乌鲁木齐830017 [2]新疆医科大学第一附属医院省部共建中亚高发病因与防治国家重点实验室 [3]新疆地方病分子生物学重点实验室 [4]新疆医科大学第五附属医院检验科 [5]新疆医科大学第三附属医院检验科

出　　处：《中国病原生物学杂志》2022年第5期520-525,531,共7页Journal of Pathogen Biology

基　　金：省部共建中亚高发病因与防治国家重点实验室开放课题(No.SKL-HIDCA-2019-10)。

摘　　要：目的探究在包虫囊液干预过程中IFITM3与Foxp3之间的相互影响。方法小鼠细粒棘球蚴病造模后收集肝脏组织,通过HE染色、免疫组织化学方法检测肝脏组织中IFITM3和Foxp3的表达状况。采用Western blot和荧光定量PCR(Real-time PCR)测定未加囊液浓度(阴性组)、1∶10、1∶100和1∶1000共4组不同囊液浓度比例干预HEK293T细胞后IFITM3和Foxp3的蛋白表达量和mRNA相对表达量。用Spearman进行IFITM3和Foxp3相关性分析。结果HE染色后,可见CE肝脏组织的病灶;免疫组化结果显示随着细粒棘球蚴感染时间的延长,IFITM3在肝脏的表达量逐渐降低,Foxp3的表达量逐渐升高。在感染第2 d IFITM3的表达最高而Foxp3的表达量最低高,感染300 d IFITM3的表达量最低,Foxp3的表达量最高。Western blot显示,在4组囊液浓度比例干预下,IFITM3的灰度值分别是(1.421±0.162)、(0.798±0.091)、(0.936±0.118)、(1.269±0.101);Foxp3的灰度值分别是(0.614±0.034)、(0.761±0.020)、(0.919±0.019)、(1.115±0.177),说明IFITM3和Foxp3随着囊液干预浓度的降低表达量逐渐升高。Real-time PCR结果显示,在4组囊液浓度比例干预后IFITM3 mRNA相对表达水平分别是(1.00±0.322)、(0.605±0.321)、(0.736±0.113)、(2.384±0.049);Foxp3 mRNA相对表达水平分别是(1.000±0.153)、(1.321±0.316)、(1.675±0.376)、(3.848±1.645),说明随着囊液干预浓度的降低,IFITM3和Foxp3mRNA的相对表达量逐渐升高。Spearman相关分析结果显示,不同浓度囊液干预后,细胞中的IFITM3与Foxp3在mRNA相对表达和蛋白表达均成正相关。结论在细粒棘球蚴感染过程中,低浓度的囊液刺激会使IFITM3和Foxp3的表达量增高,并且IFITM3与Foxp3呈正相关性,说明宿主限制性因子IFITM3通过与核转录因子Foxp3存在关联,从而与获得性免疫发生联系。Objective To explore the interaction between IFITM3 and Foxp3 under the intervention of hydatid cyst fluid.Methods The liver tissues were collected after the model of mouse echinococcosis.The expression of IFITM3 and Foxp3 in liver tissues were detected by HE staining and immunohistochemistry.Western blot(WB)and real time PCR(real time PCR)were used to determine the protein expression and mRNA relative expression of IFITM3 and Foxp3 in HEK293 T cells after intervention with different capsule concentration ratios in four groups without capsule concentration(negative group),1∶10,1∶100 and 1∶1000.Spearman was used to analyze the correlation between IFITM3 and Foxp3.Results H&E staining results showed the lesions of CE liver;Immunohistochemical results showed that with the extension of Echinococcus granulosus infection time,the expression of IFITM3 in liver gradually decreased and the expression of Foxp3 gradually increased.On the second day of infection,the expression of IFITM3 was the highest and the expression of Foxp3 was the lowest.On the 300 days of infection,the expression of IFITM3 was the lowest and the expression of Foxp3 was the highest.WB data showed that the gray values of IFITM3 were(1.421±0.162),(0.798±0.091),(0.936±0.118),(1.269±0.101)under the intervention of the concentration ratio of capsule fluid in the four groups;The gray values of Foxp3 were(0.614±0.034),(0.761±0.020),(0.919±0.019),(1.115±0.177),indicating that the expression of IFITM3 and Foxp3 increased gradually with the decrease of capsule intervention concentration.Real time PCR showed that the relative expression levels of IFITM3 mRNA were(1.00±0.322),(0.605±0.321),(0.736±0.113),(2.384±0.049)respectively;The relative expression levels of Foxp3 mRNA were(1.000±0.153),(1.321±0.316),(1.675±0.376),(3.848±1.645)respectively,indicating that the relative expression levels of IFITM3 and Foxp3 mRNA gradually increased with the decrease of capsule fluid intervention concentration.Spearman correlation analysis showed that there

关 键 词：细粒棘球蚴感染 宿主限制性因子 干扰素诱导的跨膜蛋白3 核转录因子Foxp3 

分 类 号：R383.33[医药卫生—医学寄生虫学]