PCV2与PPV共感染猪外周血单个核细胞对其细胞凋亡相关因子表达水平的影响

摘要

为分析猪圆环病毒2型(PCV2)与猪细小病毒(PPV)体外共感染对猪外周血单个核细胞(PBMC)细胞凋亡相关因子mRNA转录水平的影响,探讨PCV2和PPV共感染机制及宿主一病毒之间的作用关系,运用病毒滴度和相对荧光定量PCR技术,测定和分析PCV2和PPV感染PBMC后PCV2、PPV的病毒滴度含量及Bcl-2、FasL、p53、Cas pase-8、PBR、TNF-α等的转录时相变化.结果表明:PCV2、PPV能够感染PBMC细胞,PCV2/PPV共感染中PCV2、PPV的含量分别在24 h显著最高(P＜0.001)；PCV2、PPV单独感染和PCV2与PPV共感染PBMC后引起Bcl-2、FasL、p53、Caspase-8、PBR、TNF-α mRNA转录水平上升；在3h时PCV2/PPV共感染组PBR、P53mRNA转录水平显著高于PCV2、PPV单感染组(P＜0.05),12 h时PCV2/PPV共感染组FasL mRNA转录水平显著高于PCV2、PPV单感染组(P＜0.05),24 h时Bcl-2、Caspase-8 mRNA转录水平显著高于PCV2、PPV单感染组(P＜0.05),PCV2/PPV共感染组TNF-α mRNA转录水平显著高于PPV组(P＜0.05),与PCV2组差异不显著.结论:PCV2与PPV共感染引起细胞凋亡相关因子mRNA转录水平上调,加速淋巴细胞凋亡,本试验为PCV2和PPV共感染机制研究提供了理论基础和试验依据.%In order to investigate the influence on the mRNA expression level of apoptosis related factors in porcine peripheral blood mononuclear cells (PBMC) following the co-infection with porcine circovirus type 2 (PCV2) and porcine parvovirus (PPV) , and clarify the mechanism of PCV2 and PPV co-infection, and the interaction of the host and virus,we used PPV and PCV2 to co-infect the swine PBMC in vitro, then measured and analyzed the viral titre, and the transcript level of cytokines of Bcl-2, FasL, p53, Caspase-8, PBR and TNF-α in mRNA level using relative quantification real-time PCR. The results were as follows; PCV 2 and PPV could infect the swine PBMC in vitro, and the virus replication levels of PCV2, PPV in the PCV2/PPV co-infection group were highest 24 h post inoculation (p. i. , P<0. 001). The transcript levels of Bcl-2,FasL, p53, Caspase-8, PBR and TNF-α were increased in the PCV2, PPV and PCV2/PPV co-infection groups. The mRNA transcript levels of PBR and p53 of the PPV/PCV2 co-infected group were significantly higher than that of PCV2 or PPV infection alone at 3 h p. i. , and also with the higher expression levels of FasL in 12 h p. i. , Bcl-2 and Caspase-8 expression in 24 h p. i. (P< 0.05). Furthermore, the mRNA transcript levels of TNF-a of the PPV/PCV2 co-infected group showed significantly higher level than that of PPV infection alone (P<0. 05), but there is no significant increase when compared with PCV2 infection alone. PCV2 and PPV co-infection could increase the mRNA expression level of the apoptosis related factors, and accelerate the lymphocyte apoptosis. This research provides the basis for further study of the mechanism of PCV2 and PPV co-infection.