小反刍兽疫病毒感染外周血单个核细胞凋亡相关基因的筛选鉴定  

Identification of apoptosis-related gene in peripheral blood mononuclear cells of infected with peste des petits ruminants virus

作　　者：苏乾隆 赵帅阳[1] 李彦敏 张瑞 张志雄 孙跃峰 张志东[1,2] SU Qian-long;ZHAO Shuai-yang;LI Yan-min;ZHANG Rui;ZHANG Zhi-xiong;SUN Yue-feng;ZHANG Zhi-dong(State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;College of Animal Husbandry and Veterinary Medicine,Southwest Minzu University,Chengdu 610041,China)

机构地区：[1]中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,甘肃兰州730046 [2]西南民族大学畜牧兽医学院,四川成都610041

出　　处：《中国兽医科学》2021年第9期1140-1147,共8页Chinese Veterinary Science

基　　金：“十三五”国家重点研发计划项目(2016YFD0500108);甘肃省科技计划国际科技合作类项目(20YF3WA008)。

摘　　要：细胞凋亡、自噬、坏死及炎症反应是机体重要的抗病毒反应机制,尤其淋巴细胞凋亡是小反刍兽疫病毒(PPRV)引起宿主免疫抑制的重要因素,但其分子机制尚不清楚。本研究分别在绵羊外周血单个核细胞(PBMCs)体外感染PPRV后经24、48和72 h收样,利用PCR芯片检测共84个与凋亡、自噬、坏死及炎症有关的通路相关关键分子的变化。结果表明,感染PPRV后经24、48和72 h上调表达的基因个数分别为29、30、3,下调表达的基因个数分别为24、12、28;qPCR验证结果表明,细胞凋亡相关基因FASLG、BID、APAF1、BCL2L1、BCL2L10、CASP9、BAK1、BCL2、FAS、XIAP等随着PPRV感染时间的延长,表达量逐渐升高,CASP8随着PPRV感染时间的延长,表达量逐渐下降,与PCR芯片检测结果一致。这为进一步阐明PPRV引起的细胞凋亡及动物免疫抑制的分子机制提供理论依据。Apoptosis,autophagy,necrosis and inflammation are important antiviral mechanisms,and in particular,lymphocyte apoptosis is an important factor of host immunosuppression caused by peste des petits ruminants virus(PPRV),but its molecular mechanism had not been fully understood.In this study,sheep peripheral blood mononuclear cells(PBMCs)were infected with PPRV in vitro,and then samples were collected at 24,48 and 72 hours post infection(hpi),respectively.Subsequently,the changes of expression of key molecule genes in 84 pathways related to apoptosis,autophagy,necrosis and inflammation,were screened by PCR Array.The results showed that the numbers of up-regulated genes at 24,48 and 72 hpi were 29,30,and 3,respectively,and the numbers of down-regulated genes were 24,12,and 28,respectively.The q PCR results further showed that with the extension of infection time,the expression level of apoptosis-related genes FASLG,BID,APAF1,BCL2 L1,BCL2 L10,CASP9,BAK1,BCL2,FAS,XIAP gradually increased,in contrast,the expression level of CASP8 decreased.The q PCR results were consistent with those of PCR Array,which would help improve our understanding the molecular mechanism of apoptosis and immunosuppression caused by PPRV.

关 键 词：PCR芯片 小反刍兽疫病毒 外周血单个核细胞 凋亡 免疫抑制 

分 类 号：S852.659.5[农业科学—基础兽医学]