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为探究猪繁殖与呼吸综合征病毒(PRRSV)体外感染猪肺泡巨噬细胞(PAMs)后对Ⅰ型干扰素(IFN)及其信号分子mR NA转录的抑制效应,本研究选用PRRSV血清抗体和抗原均为阴性的4周龄健康仔猪3头,无菌分离PAMs,分为对照组和PRRSV感染组(PRRSV组),分别在培养6 h、12 h、24 h、36 h、48 h和60 h收集PAMs。并采用MTS检测各组细胞活力和荧光定量PCR检测IFN-α/β及其信号分子mR NA转录水平的动态变化。结果显示,与对照组相比PRRSV感染组细胞活力明显下降(p<0.01);IFN-αmR NA转录水平在6 h~36 h逐渐降低,36 h时与对照组相比差异极显著(p<0.01),48 h后转录水平有所上调,但仍显著低于对照组(p<0.01或p<0.05);IFN-βmR NA转录量在24 h、36 h、48 h和60 h显著低于对照组(p<0.01或p<0.05);MDA5、RIG-I、MAVS和IRF3 mR NA转录量在12 h显著下调(p<0.01)。这些结果表明,PRRSV能够导致PAMs活力降低,并通过抑制MDA5、RIG-I、MAVS和IRF3基因的转录水平从而抑制Ⅰ型IFN的产生。
In order to investigate the inhibitory effect of porcine reproductive and respiratory syndrome virus (PRRSV) on the transcription of type I interferon (IFN) and its signaling molecule mR NA after in vitro infection of porcine alveolar macrophages (PAMs), we used PRRSV serum antibody and antigen Three healthy 4-week-old healthy piglets, all of which were negative, were randomly divided into control group and PRRSV-infected group (PRRSV group), and were cultured for 6, 12, 24, 36, 48 and 60 h Collect PAMs. The activity of IFN-α / β and its mR-NA transcription were detected by MTS assay and fluorescence quantitative PCR. The results showed that compared with the control group, the viability of PRRSV infection group was significantly decreased (p <0.01); the transcriptional level of IFN-αmR NA gradually decreased from 6 h to 36 h, the difference was significant at 36 h compared with the control group (p < 0.01). After 48 h, the transcription level was increased but still significantly lower than that of the control group (p <0.01 or p <0.05). The transcript levels of IFN-β mRNA were significantly lower at 24 h, 36 h, 48 h and 60 h (P <0.01 or p <0.05). The transcript levels of MDA5, RIG-I, MAVS and IRF3 mR NA were significantly down-regulated at 12 h (p <0.01). These results indicate that PRRSV can reduce the viability of PAMs and inhibit the production of type I IFN by inhibiting the transcription of MDA5, RIG-I, MAVS and IRF3 genes.