小鼠感染BVDV诱导NLRP3炎症小体的表达及其对病毒复制的影响  

Differential expression of NLRP3inflammasome induced by BVDV infection in mice and its effect on virus replication

作　　者：刘思雨 张月莹 黄江 崔月琦 刘宇[1,2] 周玉龙 朱战波[1,2] 张泽财 LIU Siyu;ZHANG Yueying;HUANG Jiang;CUI Yueqi;LIU Yu;ZHOU Yulong;ZHU Zhanbo;ZHANG Zecai(College of Animal Science and Veterinary Medicine,Heilongjiang Bayi Agricultural University,Daqing 163319,China;Heilongjiang Provincial Technology Innovation Center for Bovine Disease Control and Prevention,Heilongjiang Bayi Agricultural University,Daqing 163319,China)

机构地区：[1]黑龙江八一农垦大学动物科技学院,黑龙江大庆163319 [2]黑龙江省牛病防控工程技术研究中心,黑龙江大庆163319

出　　处：《中国农业大学学报》2022年第10期134-141,共8页Journal of China Agricultural University

基　　金：国家自然科学基金项目(32072896);黑龙江省博士后落户科研启动金(2032030150);黑龙江八一农垦大学研究生创新科研项目(YJSCX2021-Y16)。

摘　　要：为分析小鼠感染牛病毒性腹泻病毒(Bovine viral diarrhea virus,BVDV)诱导的NLRP3炎症小体表达情况及其对病毒复制的影响,选用40只4~6周龄雌性SPF级BALB/c小鼠,随机分成8组,其中BVDV接种小鼠7组,对照1组,每组5只。应用105 TCID50的CP型BVDV腹腔感染小鼠后,分别在感染后0、6、12、24、48、96、168和240h采集小鼠的肠道、脾脏和血液,通过qRT-PCR和Western Blot方法对小鼠十二指肠中NLRP3、IL-1β和Caspase-1分别进行基因及蛋白水平的检测。应用qRT-PCR方法确定不同时间点小鼠十二指肠和血液中的病毒载量,同时于感染后第168h检查十二指肠和脾脏病理组织变化。结果表明:1)小鼠感染BVDV早期(6、12和24h)组织中的NLRP3、IL-1β和Caspase-1基因转录水平逐步升高,48h未见明显变化,同时感染24h时,蛋白表达水平均显著高于对照组(P<0.05);2)小鼠感染BVDV后期(96、168和240h)NLRP3、IL-1β和Caspase-1的基因转录水平呈现下降趋势,168h时转录和蛋白水平降低最为显著(P<0.05);3)感染小鼠十二指肠、脾脏和血液的病毒载量均在168h达到高峰;第168h采集的小鼠十二指肠有大量炎性细胞浸润,脾脏淋巴细胞变性和坏死,证明BVDV感染模型成立。综上,NLRP3炎症小体在BVDV感染早期被激活,然而,随着病毒载量的增加,可能对其活性有一定的抑制作用。The aim of this study was to analyze the activation of NLRP3 inflammasome induced by Bovine viral diarrhea virus(BVDV)infection in mice and its influence on virus replication.Forty female SPF BALB/c mice aged 4-6weeks were randomly divided into eight groups,including seven groups inoculated with BVDV and one control group with five mice in each group.After intraperitoneal infection of mice with CP BVDV of 105 TCID50,the intestine,spleen and blood were collected at 0,6,12,24,48,96,168and 240h,respectively.The expressions of NLRP3,IL-1βand Caspase-1were detected by qRT-PCR and Western blot.The viral loads in duodenum and blood at different time points were detected by qRT-PCR,and the pathological changes of duodenum and spleen were examined at 168hafter infection.The results showed that:1)The mRNA expressions of NLRP3,IL-1βand Caspase-1in the early stage(6,12and 24h)of BVDV infection increased gradually and there was no significant change at 48h.Simultaneously,the protein expression level was significantly higher than those in the control group at 24hof infection(P<0.05);2)In the late stage of BVDV infection(96,168and 240h),the expression of NLRP3,IL-1βand Caspase-1 mRNA showed a downward trend,and the transcription and protein levels decreased most significantly at 168h(P<0.05);3)The viral loads in duodenum,spleen and blood of infected mice reached the peak at 168h;There were a large number of inflammatory cell infiltration in the duodenum,ymphocyte degeneration and necrosis were observed in the spleen,which proved that the BVDV infection model was established.In conclusion,NLRP3inflammasome in the early stage of BVDV infection was activated.However,with the increase of virus load,it may inhibit its activity.

关 键 词：牛病毒性腹泻病毒 BALB/C小鼠 NLRP3炎症小体 病毒载量 

分 类 号：S85[农业科学—兽医学]