摘要
为了研究伪狂犬病毒(PRV)感染对小鼠脾脏细胞周期的影响及相关因子的变化,本研究分别以1×10^(3)TCID_(50)/100μL、1×10^(4)TCID_(50)/100μL和1×10^(5)TCID_(50)/100μL剂量经皮下接种感染小鼠48 h后采集小鼠脾脏,采用流式细胞检测法对脾脏细胞周期进行检测,结果显示,和对照组比较,不同剂量PRV感染小鼠48 h后,G0/G1期脾脏细胞比率均显著增加,处于S期和G2期的脾脏细胞均显著减少,且各剂量组间无差异。提取低剂量组小鼠脾脏RNA,以RT-qPCR检测细胞周期相关基因转录水平,结果显示,和对照组比较,CDK4和Cyclin D基因转录水平显著下调;提取低剂量组小鼠脾脏总蛋白,经western blot检测分析上述两个基因蛋白的表达水平,结果显示,和对照组比较,CDK4和Cyclin D蛋白表达水平均降低。本研究首次证实PRV感染小鼠后可通过下调表达细胞周期相关因子CDK4和Cyclin D,将脾脏细胞周期阻滞在G0/G1期,为进一步研究PRV免疫抑制机理提供一定的数据参考。
To investigate the effect of pseudorabies virus(PRV)on spleen cell cycle and its related factors,the mice were inoculated subcutaneously with PRV a dose of 1×10^(3)TCID_(50),1×10^(4)TCID_(50)and 1×10^(5)TCID_(50),respectively.Compared with the control group,the each experimental group results of flow cytometry showed that at 48h post infection,the spleen cells in the G0/G1 phase were significantly increased,while the spleen cells in S phase and G2 phase were significantly decreased,but there was no difference among the dose groups.In this experiment.Spleen RNA was extracted from mice in low dose group,and the transcription levels of cell cycle related-genes were detected by quantitative RT-PCR method.The results showed that the transcription levels of CDK4 and Cyclin D were significantly down-regulated.In addition,the total spleen protein in low dose group was extracted and analyzed by western blot.The results showed that the protein expression levels of CDK4 and Cyclin D were also significantly down-regulated.The results of this study confirmed for the first time that PRV infection could arrest the splenic cell cycle at G0/G1 phase by decreasing the expression of cell cycle-related factors of CDK4 and cyclin D.This study should offer some information for further study on the immunosuppressive mechanism of PRV.
作者
刘杉杉
孙伟
周佳
曾泽
罗杰
杨慧
张稳
LIU Shan-shan;SUN Wei;ZHOU Jia;ZENG Ze;LUO Jie;YANG Hui;ZHANG Wen(College of Agriculture,Tongren Polytechnic College,Tongren 554300,China;National and Local Engineering Research Centre for Separation and Purification Ethnic Chinese Veterinary Herbs,Tongren 554300,China)
出处
《中国预防兽医学报》
CAS
CSCD
北大核心
2021年第10期1085-1088,共4页
Chinese Journal of Preventive Veterinary Medicine
基金
贵州省科技厅科技计划项目(黔科合基础[2019]1456号)
铜仁市科技局计划项目(铜市科研[2019]12-6)。
