苦参碱对PRRSV的体外抑制作用及其机制

Inhibitory action and mechanism of matrine against porcine reproductive and respiratory syndrome virus in vitro

本试验以Marc-145细胞为模型,采用细胞病变(CPE)抑制试验和MTT法测定细胞活力,观察苦参碱(ma-trine)体外抗猪繁殖呼吸与综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)活性,并从抑制病毒复制、阻断吸附和直接杀灭3个方面探讨苦参碱体外抑制PRRSV的作用机制。结果显示,苦参碱CC50>1 500mg/L,最大安全浓度为750mg/L,EC50为(443.5±33.4)mg/L,SI≥3.38,在安全浓度范围内对PRRSV感染Marc-145细胞的抑制率达到93.6%;在直接杀灭和复制抑制试验中最高抑制率分别达到105.10%和91.53%;在病毒吸附抑制试验中抑制率<20%,表明苦参碱不能阻断PRRSV对Marc-145细胞的吸附。结果表明,苦参碱在体外可有效抑制PRRSV对Marc-145细胞的感染,其抗病毒机制可能是干扰病毒的复制或/和直接杀灭病毒,提示苦参碱可作为研发预防及治疗猪蓝耳病的药物或先导化合物。

To observe the inhibitory action of matrine against PRRSV in vitro using visualization of cytopathologic effect （CPE） assay and MTT test on Marc-145 cell,and explores the mechanism by time-of-addition assay,adsorption inhibition assay and virucidal assay. The result shows that CCs0 of matrine is larger than 1 500 mg/L. The maximum non-cytotoxic concentration （MNTC） and EC0 are nearly 750 mg/L and （443.5±33.4） mg/L separately,S/is greater than 3.8,the inhibi- tion ratio of Matrine against PRRSV up to 93.6% in MNTC. The inhibition ratios separately are up to 105.10% and 91.53% in virucidal assay and the time-of-addition assay,lower than 20% of the inhibition ratios in the adsorption inhibition assay showes that matrine did not inhibite virus attach to Marc-145 cell. The experimental results demonstrates that matrine could reduce the damage of PRRSV to Marc-145 cell effectively. Its mechanism of antiviral actions could be due to inhibit the virus in the early or middle stage of replication or / and inactivate the virus directly, which suggests that matrine could be used for developing a new drug or precursor in treating and preventing porcine reproductive and respiratory syndrome.