包虫囊液干预的肝细胞Foxp3与IFITM3表达变化及其与干扰素刺激蛋白相关性分析  

Expression changes of Foxp3 and IFITM3 in liver cells intervened by hydatid cyst fluid and their correlations with interferon-stimulated proteins

作　　者：杨雨阳 蒙克嘎勒 阿不来提·阿合买提 屈晴[1] 齐新伟 单骄宇[1,2,3] YANG Yuyang;Mengkegale;Abulaiti·Ahemaiti;QU Qing;QI Xinwei;SHAN Jiaoyu(Department of Human Parasitology,College of Basic Medicine of Xinjiang Medical University,Urumqi 830000,China;不详)

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,乌鲁木齐830000 [2]新疆医科大学第一附属医院中亚高发疾病发病机制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]新疆医科大学基础医学院生理学教研室 [5]新疆医科大学基础医学院机能中心实验室

出　　处：《山东医药》2022年第17期1-6,共6页Shandong Medical Journal

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项目(SKL-HIDCA-2019-10)。

摘　　要：目的观察包虫囊液干预的肝细胞核转录因子Foxp3、干扰素诱导跨膜蛋白3(IFITM3)表达变化并分析其与干扰素刺激蛋白IFITM1、IFITM2、IFIT3、ISG15的相关性。方法从包虫病羊肝脏包囊中抽取囊液,将人肝癌细胞HepG2分为高浓度组、中浓度组、低浓度组及对照组,分别加入1∶10、1∶100和1∶1000浓度比例的囊液及不加囊液,采用Western blotting法及荧光定量PCR法检测各组干预后及低浓度组在干预后0、12、24、36、48、60、72 h细胞中的Foxp3、IFITM3及相关干扰素刺激蛋白IFITM1、IFITM2、IFIT3、ISG15蛋白及mRNA表达,采用Spearman分析Foxp3、IFITM3与IFITM1、IFITM2、IFIT3、ISG15的相关性。结果不同囊液浓度干预下,IFITM1、IFITM2、IFITM3、Foxp3、ISG15蛋白及mRNA表达随着干预囊液浓度降低逐渐升高,IFIT3蛋白表达中浓度组>低浓度组、高浓度组(P均<0.05),IFIT3 mRNA高、中、低浓度组差异无统计学意义。低囊液浓度干预下各时点IFITM1、IFITM2、IFIT3蛋白表达随囊液干预时间延长而降低,IFITM3、ISG15蛋白表达随囊液干预时间延长先升高后降低,Foxp3蛋白表达随囊液干预时间延长而升高;IFITM2、IFIT3、ISG15 mRNA表达随囊液干预时间延长而降低,IFITM3、IFITM1 mRNA表达随囊液干预时间延长先升高后降低,Foxp3 mRNA表达随囊液干预时间延长而升高。Spearman相关分析结果显示,不同囊液浓度干预下Foxp3表达与IFITM3表达呈正相关(P=0.004);Foxp3 mRNA、IFITM3蛋白及mRNA表达与IFITM1、IFITM2、ISG15 mRNA表达均呈正相关(P均<0.05),与IFIT3 mRNA表达无相关性;Foxp3蛋白表达与IFITM1、ISG15蛋白表达呈正相关(P均<0.05),与IFITM2和IFIT3蛋白表达无相关性。低囊液浓度干预下各时点Foxp3表达与IFITM3表达呈负相关(P均<0.05);Foxp3表达与IFITM1、IFITM2、IFIT3、ISG15表达呈负相关;IFITM3表达与IFITM1、IFITM2、IFIT3、ISG15表达呈正相关(P均<0.05)。结论在低浓度包虫囊液干预下,HepG2细胞IObjective To observe the expression changes of hepatic nuclear transcription factor Foxp3 and interferon-induced transmembrane protein 3(IFITM3)in liver cells intervened by hydatid cyst fluid,and to analyze their correlations with interferon-stimulated proteins IFITM1,IFITM2,IFIT3,and ISG15.Methods The cyst fluid was extracted from the liver cyst of hydatid sheep.Human hepatoma cells HepG2 were divided into the high-concentration group,medium-concentration group,low-concentration group,and control group;cells in the first three groups were added with capsule solutions in the concentration ratios of 1∶10,1∶100 and 1∶1000,and the cells in the control group were not added.The expression levels of Foxp3,IFITM3,and the related interferon-stimulated proteins IFITM1,IFITM2,IFIT3 and ISG15 protein and mRNA in cells of each group and of the low concentration group at 0,12,24,36,48,60 and 72 h after intervention were detected by Western blotting and fluorescent quantitative PCR.Spearman was used to analyze the correlations between Foxp3,IFITM3 and IFITM1,IFITM2,IFIT3,and ISG15.Results Under the intervention of different capsule concentration,the protein and mRNA expression levels of IFITM1,IFITM2,IFITM3,Foxp3,and ISG15 increased gradually with the decrease of intervention capsule concentration.The expression of IFIT3 protein was higher in the medium-concentration group than in the low-concentration group and high-concentration group(both P<0.05).There was no significant difference in the IFIT3 mRNA among the high-,medium-and low-concentration groups.After the lowconcentration intervention,the protein expression levels of IFITM1,IFITM2,and IFIT3 decreased over the course of cyst fluid intervention at each time point,while the protein expression levels of IFITM3 and ISG15 first increased and then decreased over the course of cyst fluid intervention,and the expression of Foxp3 protein increased with the cyst fluid intervention time.The mRNA expression levels of IFITM2,IFIT3 and ISG15 decreased over the course of cyst fl

关 键 词：包虫囊液 干扰素诱导跨膜蛋白3 核转录因子Foxp3 细粒棘球蚴感染 干扰素刺激基因 

分 类 号：R383[医药卫生—医学寄生虫学]