| 本文已被：浏览 941次 下载 1250次 |
码上扫一扫！ |
| 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 |
|
刘思雨1,张月莹1,黄江1,崔月琦1,刘宇1,2,周玉龙1,2,朱战波1,2*,张泽财1,2*
|
|
|
| (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) |
|
| 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240 h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168 h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24 h)组织中的NLRP3、IL-1β和Caspase-1基因转录水平逐步升高，48 h未见明显变化，同时感染24 h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240 h)NLRP3、IL-1β和Caspase-1的基因转录水平呈现下降趋势，168 h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168 h达到高峰；第168 h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。 |
| 关键词: 牛病毒性腹泻病毒 BALB/c小鼠 NLRP3炎症小体 病毒载量 |
| DOI：10.11841/j.issn.1007-4333.2022.10.12 |
| 投稿时间：2022-01-09 |
| 基金项目: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) |
|
| Differential expression of NLRP3 inflammasome induced by BVDV infection in mice and its effect on virus replication |
|
LIU Siyu1,ZHANG Yueying1,HUANG Jiang1,CUI Yueqi1,LIU Yu1,2,ZHOU Yulong1,2,ZHU Zhanbo1,2*,ZHANG Zecai1,2*
|
| (1.College of Animal Science and Veterinary Medicine, Heilongjiang Bayi Agricultural University, Daqing 163319, China;2.Heilongjiang Provincial Technology Innovation Center for Bovine Disease Control and Prevention, Heilongjiang Bayi Agricultural University, Daqing 163319, China) |
| Abstract: |
| 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 4-6 weeks 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, 168 and 240 h, respectively. The expressions of NLRP3, IL-1β and Caspase-1 were 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 168 h after infection. The results showed that: 1)The mRNA expressions of NLRP3, IL-1β and Caspase-1 in the early stage(6, 12 and 24 h)of BVDV infection increased gradually and there was no significant change at 48 h. Simultaneously, the protein expression level was significantly higher than those in the control group at 24 h of infection(P<0. 05); 2)In the late stage of BVDV infection(96, 168 and 240 h), the expression of NLRP3, IL-1β and Caspase-1 mRNA showed a downward trend, and the transcription and protein levels decreased most significantly at 168 h(P<0. 05); 3)The viral loads in duodenum, spleen and blood of infected mice reached the peak at 168 h; 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, NLRP3 inflammasome in the early stage of BVDV infection was activated. However, with the increase of virus load, it may inhibit its activity. |
| Key words: BVDV BALB/c mice NLRP3 inflammatory virus load |
