猪繁殖与呼吸综合征病毒在猪动脉血管内皮细胞上的培养特性

Experimental studies on the culture of porcine reproductive and respiratory syndrome virus in porcine arterial endothelial cells

探索应用原代猪血管内皮细胞培养猪繁殖与呼吸综合征病毒(PRRSV)的方法,为进一步研究PRRSV对血管内皮细胞的损伤及其作用机制建立模型。用Ⅱ型胶原酶灌注60日龄健康仔猪颈总动脉分离获得动脉血管内皮细胞,以PRRSVHuN4株接种原代猪血管内皮细胞,进行细胞形态和荧光定量RT-PCR检测。结果显示,PRRSV接种原代猪血管内皮细胞后,呈现典型的细胞病变,病毒增殖规律与Marc-145细胞基本相同。结果表明,在猪血管内皮细胞上成功复制了PRRSV,为下一步研究PRRSV在血管内皮细胞中的复制及其导致的病理损伤提供了试验依据。

The main purpose of this study was to establish the culture method for porcine reproductive and respiratory syndrome virus（PRRSV）in primary porcine arterial endothelial cells and to establish cell model of injury in PRRSV for further investigation of its mechanism.Collagenase typeⅡ was used to isolate endothelial cells from the carotid artery of 60-day-old healthy piglets by irrigative digestion technique. The cultured primary porcine arterial endothelial cells inoculated by PRRSV HuN4 strain and control cells were observed by optical microscopy and the concentrations of PRRSV RNA were detected by quantitative polymerase chain reaction.The marked cytopathological alterations were observed in cells inoculated by PRRSV.The growth curves of PRRSV HuN4 strain showed the similar laws in primary porcine arterial endothelial cells and Marc-145 cells.The method of culturing PRRSV was first reported in primary porcine arterial endothelial cells,and the results showed that this study provided one experimental evidence that multiplication of PRRSV in porcine endothelial cells could cause the CPE.