论文部分内容阅读
目的观察硒蛋白对手足口病病原体肠道病毒71型(EV71)在感染动物体内复制的影响情况。方法将30只雌性ICR小鼠分为常规饲料组、缺硒饲料组和加硒饲料组,各种饲料饲养小鼠6周后,通过电感耦合等离子体质谱仪(ICP-MS)测定小鼠外周血中硒的含量以及荧光定量PCR方法测定动物体内肠道病毒EV71的载量,然后用病毒空斑实验测定病毒滴度并用酶促法来测定主要硒蛋白酶的活性。结果通过测定外周血中硒的含量发现,缺硒饲料组的ICR小鼠硒含量为0.081μg/g,显著低于常规饲料组0.135μg/g和加硒饲料组的0.128μg/g,差异有统计学意义(P<0.01);用荧光定量PCR方法测定EV71载量,加硒饲料组ICR小鼠体内EV71核酸扩增Ct值,明显高于缺硒饲料组(P<0.05);ICR小鼠体内主要硒蛋白酶(GPX1)的活力在缺硒饲料组的动物体内显著低于加硒饲料组;病毒空斑实验结果表明,缺硒饲料组的ICR小鼠体内EV71病毒复制活力显著高于其他两组。结论缺硒可能导致硒蛋白GPX1合成减少或其活力降低从而加快EV71在宿主内的复制。
Objective To investigate the effect of selenoprotein on the replication of enterovirus 71 (EV71), a pathogen of foot-mouth disease in animals. Methods Thirty female ICR mice were divided into the normal diet group, the selenium-deficient diet group and the selenium-fed diet group, and the mice were fed with various feedstuffs for 6 weeks. The peripheral blood samples were determined by inductively coupled plasma mass spectrometry (ICP-MS) Blood selenium content and fluorescence quantitative PCR method was used to determine the content of enterovirus EV71 in animals. The viral titer was determined by virus plaque assay and the activity of major seial protease was determined by enzymatic method. Results Selenium content in peripheral blood was found to be 0.081 μg / g in ICR mice fed selenium deficient diet, which was significantly lower than 0.135 μg / g in normal diet group and 0.128 μg / g in selenium fed group (P <0.01). The quantitative RT-PCR method was used to determine the EV71 load, and the Ct value of EV71 in sera of ICR mice fed selenium diet was significantly higher than that of selenium-deficient diet (P <0.05) The activity of main seial protease (GPX1) in the selenium-deficient diet group was significantly lower than that of the selenium-fed group. The virus plaque test results showed that the replication activity of EV71 virus in the selenium-deficient diet group was significantly higher than that of the other two group. Conclusions Selenium deficiency may result in the decrease of selenoprotein GPX1 synthesis or decrease of its vitality and thus accelerate the replication of EV71 in the host.