天然中草药提取物抗猪繁殖与呼吸综合征病毒活性研究

Study on Antiviral Activity of Four Chinese Herbs to Porcine Reproductive and Respiratory Syndrome Virus

为研究天然中草药对猪繁殖与呼吸综合征病毒(PRRSV)的抗病毒活性,本研究以体外细胞培养体系为模型,在采用CCK-8法测定青黛(Indigo naturalis)、花椒(Zanthoxylum bungeanum)、卫矛(Euonymus ala-tus)和沉香(Aquilaria agallocha Roxb.)4种中草药提取物对细胞的半数毒性浓度(TC50)及最大无毒浓度(TC0)基础上,通过观察细胞病变效应(CPE),实时荧光定量RT-PCR方法检测病毒核蛋白基因(ORF7)mRNA表达量的变化,对天然中草药提取物抑制PRRSV增殖能力进行评价。实验结果分析显示:青黛、花椒、卫矛和沉香4种不同提取物的药物抗病毒选择指数(SI)分别为10.3,5.2,12.7,6。实时荧光检测病毒基因mRNA表达量变化结果表明:这4种天然中草药提取物在一定浓度范围内均能显著(P<0.05)抑制病毒NP蛋白基因(ORF7)的表达。本次研究结果证实,4种受试中草药提取物对PRRSV的增殖均具有显著(P<0.05)的抑制效果,以卫矛提取物的效果最佳。

To study the antiviral activity of natural herbal to porcine reproductive and respiratory syndrome virus（PRRSV）,antiviral effect of Indigo naturalis,Zanthoxylum bungeanum,Euonymus alatus and Aquilaria agallocha Roxb.were analyzed by using in vitro cell culture system model.Based on detection of median toxic concentration（TC50） and maximal nontoxic concentration（TC0） of four herbal extracts by CCK-8 method,the cytopathic effect was observed and virus nucleoprotein gene（ORF7） mRNA expression was determined by real-time quantitative RT-PCR.Results of cytotoxicity and virus proliferation inhibition analysis showed that antiviral selection index（SI） of I.naturalis,Z.bungeanum,E.alatus and L.squilariae resinatum were 10.3,5.2,12.7 and 6 respectively.Besides,mRNA expression level of viral NP protein gene（ORF7） determined by real-time PCR were significantly inhibited within certain range of concentration（P〈0.05）.Results of this study confirm that extracts of four herbals have obvious anti-PRRSV effects,the best effect found in extract of Euonymus Alatus.