ChMDA5通路参与传染性法氏囊病病毒致雏鸡法氏囊损伤的研究  

Study on the Involvement of Chicken Melanoma Differentiation-Associated Gene 5 Pathway in Bursa Injury of Chicken Induced by Infectious Bursal Disease Virus

作　　者：栾亚男 许丹蕾 葛铭 唐泽群 赵霞[1,2] 张瑞莉 LUAN Ya’nan;XU Danlei;GE Ming;TANG Zequn;ZHAO Xia;ZHANG Ruili(College of Veterinary Medicine,Northeast Agricultural University,Harbin 150030,China;Heilongjiang Key Laboratory for Laboratory Animals and Comparative Medicine,Harbin 150030,China)

机构地区：[1]东北农业大学动物医学学院,哈尔滨150030 [2]黑龙江省实验动物与比较医学重点实验室,哈尔滨150030

出　　处：《畜牧兽医学报》2020年第10期2557-2566,共10页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：国家自然科学基金(31272533)。

摘　　要：传染性法氏囊病病毒(IBDV)为dsRNA病毒,可造成雏鸡法氏囊损伤进而发生免疫抑制;MDA5是能够特异性识别dsRNA病毒的模式识别受体。为研究鸡MDA5(chMDA5)信号通路在IBDV致雏鸡法氏囊病理损伤中的作用,试验选取50只14日龄SPF雏鸡随机分为IBDV感染组和空白对照组,每组25只,IBDV感染组雏鸡通过点眼、滴鼻感染IBDV JIC7株病毒液,0.6 mL·只^-1,空白对照组雏鸡经相同途径给予相同剂量无菌PBS,感染IBDV后第1、4、7、21及35天采集雏鸡法氏囊。采用qRT-PCR方法检测法氏囊中IBDV载量,chMDA5及chMDA5信号通路衔接蛋白(chIPS-1)、转录因子(chIRF3和chNF-κB)、下游产物细胞因子(chIFN-β、chTNF-α、chIL-1β、chIL-6) mRNA水平变化;间接免疫荧光法检测chMDA5蛋白表达变化,传统病理学方法检查法氏囊病理组织学变化。结果发现,雏鸡感染IBDV后,其法氏囊中chMDA5、chIPS-1、chIRF3、chNF-κB、chIFN-β、chTNF-α、chIL-1β、chIL-6的表达量均显著高于对照组,且法氏囊组织发生形态损伤,上述变化趋势与IBDV载量变化基本一致。结果表明,雏鸡法氏囊chMDA5及其信号转导通路可被IBDV激活,参与到IBDV感染雏鸡法氏囊损伤与抗损伤过程中。Infectious bursal disease virus(IBDV),a double-stranded RNA virus,can cause the infection of chickens with bursal lesions,leading to immunosuppression;melanoma differentiation-associated gene 5(MDA5)is a pattern recognition receptor that specifically recognises double-stranded RNA viruses.In order to investigate the mechanism of the chicken MDA5(chMDA5)signalling pathway in pathological injury in IBDV-infected chickens,fifty 14-day-old specific pathogen-free(SPF)chickens were allocated into 2 groups(infected and control)and twenty-five chickens were in each group.Each chicken of infected group was inoculated with 0.6 mL IBDV JIC7 virus solution intraocularly and intranasally.The chickens in control group were administered sterile phosphate buffered saline(PBS)in the same manner simultaneously.The chickens’bursa of Fabricius was isolated at the 1st,4th,7th,21st and 35th day after infection.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect the IBDV load,and mRNA expression of chMDA5,chMDA signal pathway adaptor protein(chIPS-1),transcription factor(chIRF 3,chNF-κB)and downstream product cytokines(chIFN-β,chTNF-α,chIL-1βand chIL-6).Indirect immunofluorescence assay was performed for the expression of chMDA5 protein in chicken bursa of Fabricius.Conventional pathological methods were used to examine the pathological changes in the bursa of Fabricius.The results showed that the expression of chMDA5 as well as chIPS-1,chIRF3,chNF-κB,chIFN-β,chTNF-α,chIL-1βand chIL-6 were significantly higher than those in the control group,and morphological damage occurred in the bursal tissue in the infected group.The aforementioned trend was consistent with the change in IBDV load.The results indicated that chMDA5 and its signal transduction pathway could be activated by IBDV,which involves the bursa of Fabricius injury and anti-injury process in chickens infected with IBDV.

关 键 词：IBDV 法氏囊 chMDA5信号转导 病理形态损伤 雏鸡 

分 类 号：S852.659.4[农业科学—基础兽医学]