JAK-STAT信号通路JAK2、STAT3和SOCS1基因在鸭肠炎病毒感染中各组织的表达变化

Changes of expression of JAK2,STAT3and SOCS1genes in JAK-STAT signaling pathway in various tissues of duck infected with duck enteritis virus

将100只健康30日龄雏鸭随机分为试验组(Ⅰ组,50只)和对照组(Ⅱ组,50只),进行鸭肠炎病毒(duck enteritis viral,DEV)检测。试验组每只雏鸭腿部肌肉注射0.2 mL(1000LD_(50)),Ⅱ组注射无菌生理盐水0.2 mL,分别于感染后12,24,36,48,72,84,96 h无菌采集各组织样本(肌肉、胸腺、十二指肠、法氏囊、心脏、肝脏、脾脏、肺脏、肾脏、脑),利用荧光定量PCR和ELISA检测方法确定Ⅱ组未发生感染、Ⅰ组成功感染后,所有样品-80℃保存备用。同时,对各时段各组织中JAK2、STAT3和SOCS1基因的转录水平与蛋白质表达量进行检测,并对十二指肠进行转录水平和蛋白表达水平验证。结果显示,与Ⅱ组相比,JAK2、STAT3和SOCS1基因在DEV感染鸭机体组织中转录水平差异显著(P<0.05)。JAK2和STAT3蛋白表达水平低于Ⅱ组,差异显著(P<0.05),而SOCS1蛋白表达量水平高于Ⅱ组,差异显著(P<0.05);取十二指肠组织进行转录组测序和Western blot验证,结果与荧光定量PCR和ELISA检测结果一致。结果表明,感染DEV对雏鸭各组织JAK-STAT信号通路相关基因JAK2、STAT3和SOCS1基因的转录水平均有不同程度的影响。感染DEV对鸭机体组织JAK-STAT信号通路SOCS1基因的影响是在蛋白水平的上调,而非转录水平,同时SOCS1基因对JAK2、STAT3基因的抑制作用也是在蛋白水平。

The purpose of this study is to explore the changes in the transcription level and protein expression of JAK2,STAT3 and SOCS1 genes related to the JAK-STAT signaling pathway in various tissues of duck infected with duck enteritis virus(DEV).One hundred healthy 30-day-old ducklings were randomly divided into experimental group and control group,50 ducklings in each group.Each duck in the experimental group was injected with 0.2 mL DEV(1000 LD_(50))in the leg,ducks in the control group was injected with 0.2 mL physiological saline.Tissue samples(muscle,thymus,duodenum,bursa of Fabricius,heart,liver,spleen,lung,kidney,brain)were aseptically collected 12,24,36,48,72,84 and 96 hafter infection.Fluorescence quantitative PCR and ELISA were used to detect the transcription level and protein expression of JAK2,STAT3 and SOCS1 genes in each tissue at each time period,and the transcription level was further verified by transcriptome sequencing.The results showed that the mRNA expression level of JAK2 gene did not increase significantly after infection with DEV,while the expression of JAK2 protein was lower than that of the control group,it was significantly inhibited.The transcription levels of STAT3 and JAK2 genes showed consistency in tissue and time,while the expression of STAT3 protein was lower than that of the control group,which was inhibited.The transcription level of SOCS1 gene did not appear to be significantly up-regulated,but the protein expression level of SOCS1 was higher than that of the control group,and there was a significant increase.The above results indicate that DEV infection has different effects on the transcription levels of JAK2,STAT3 and SOCS1 genes related to the JAK-STAT signaling pathway in various tissues of ducklings,and the expression levels of JAK2 and STAT3 proteins can up-regulate the expression of SOCS1 protein.After infection with DEV,SOCS1 gene inhibits JAK2 and STAT3 genes at the protein level,not at the transcription level,and its own upregulation is also at protein level,not transcription level.