硫酸化茯苓多糖对PRRSV体外感染Marc-145细胞作用的影响

Effect of Sulfated Pachymaran Polysaccharide on PRRSV Infected Marc-145 in vitro

用稀碱法从茯苓中提取茯苓多糖,应用氯磺酸-吡啶法得到硫酸化茯苓多糖。通过观察细胞病变(CPE)效应来评价不同浓度硫酸化茯苓多糖和不同作用方式对猪繁殖与呼吸综合征病毒(PRRSV)感染Marc-145细胞的影响。结果表明,硫酸酯化茯苓多糖预作用于单层细胞12 h后感染104TCID50PRRSV,能明显阻断病毒感染细胞,其阻断病毒感染作用的最小浓度为100 mg/L;而硫酸化茯苓多糖预作用4 h后感染104TCID50的PRRSV阻断感染作用不明显,显示硫酸化茯苓多糖体外对PRRSV感染细胞的阻断感染作用呈一定的浓度和时间依赖性。感染剂量为104TCID50PRRSV后加入不同浓度硫酸化茯苓多糖,也能有效抑制病毒在细胞上的感染与增殖,其作用最小浓度为50 mg/L;而不同浓度多糖与104TCID50病毒在体外共同孵育12 h后加入到单层细胞,多糖直接杀灭PRRSV的最小浓度为100 mg/L。

The pachymaran polysaccharide was extracted by alkaline solution and then modified by chlorosulfonic-pyridine method to obtain sulfated pachymaran polysaecharide （SPP）. The effects of SPP on growth of Mark-145 cells infected with PRRSV were investigated by observing eytopathie effect （CPE）. The results showed that infecting 104TCID50 PRRSV on Mark-145 which was pre-treated of SPP for 12 h, the antiviral effect was abvious, and the minimal inhibitory concentration （MIC） was 100 mg/L. And if the pre-treation time shorted from 12 h to 4 h, the antiviral effect decreased. Adding different concentration of SPP after PRRSV infection, the infection and duplication of PRRSV was inhibited effectively and the MIC was 50 mg/L. Adding the complex of different concentration SPP and PRRSV which co-cultured 12 h in vitro to Mark-145, MIC of SPP was 100 mg/L. The results indicated that anti-PRRSV effect of SPP showed a dose or time dependent manner.