诃子、矮紫堇、甘青乌头提取物对牛病毒性腹泻病毒的体外抑制作用

doi:10.11843/j.issn.0366-6964.2018.09.025

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (9): 2036-2043.doi: 10.11843/j.issn.0366-6964.2018.09.025

诃子、矮紫堇、甘青乌头提取物对牛病毒性腹泻病毒的体外抑制作用

王丹阳, 张康, 王旭荣, 王海瑞, 王磊, 张凯, 张景艳, 李建喜*, 王学智*

中国农业科学院兰州畜牧与兽药研究所, 甘肃省新兽药工程重点实验室, 兰州 730050

收稿日期:2017-12-06 出版日期:2018-09-23 发布日期:2018-09-23
通讯作者: 王学智(1969-),男,甘肃静宁人,研究员,博士,主要从事兽医临床诊断的研究,E-mail:wangxz628@sina.com;李建喜(1971-),男,甘肃庄浪人,研究员,博士,主要从事兽医临床诊断的研究,E-mail:fyjohn@sina.com
作者简介:王丹阳(1991-),女,黑龙江人,硕士生,主要从事临床兽医学研究,E-mail:2434370881@qq.com
基金资助:
防治螨病和痢疾藏中兽药制剂制备关键技术研究与应用（201303040-14）；国家奶牛产业技术体系科学家岗位项目（CARS37）；中国农业科学院科技创新工程（CAAS-ASTIP-2017-LIHPS）

Inhibitory Effects of Extracts from Terminalia chebula, Corydalis hendersonii, Aconitum tanguticum on Bovine Viral Diarrhea Virus in vitro

WANG Dan-yang, ZHANG Kang, WANG Xu-rong, WANG Hai-rui, WANG Lei, ZHANG Kai, ZHANG Jing-yan, LI Jian-xi*, WANG Xue-zhi*

Key Labaratory of New Animal Drug Project of Gansu Province, Lanzhou Institute of Husbandry and Pharmaceutical Sciences of CAAS of Ministry of Agriculture, Lanzhou 730050, China

Received:2017-12-06 Online:2018-09-23 Published:2018-09-23

摘要/Abstract

摘要：

为探讨诃子、矮紫堇、甘青乌头3种藏药的体外抗牛病毒性腹泻病毒（BVDV）作用效果，利用细胞培养技术，对BVDV进行增殖并检测TCID₅₀；通过CPE观察和CCK8检测相结合的方法测定3种藏药和利巴韦林的最大药物安全浓度和药物半数中毒浓度（CC₅₀）；采用先加药后加病毒、先加病毒后加药、药和病毒预先作用的3种不同加药方式进行体外抗病毒抑制试验，计算不同作用方式下抗病毒有效率、药物半数有效浓度（EC₅₀）和治疗指数（TI），进一步明确各个药物对BVDV的抑制作用。结果表明，经Reed-Muench氏法计算出病毒的TCID₅₀为10^-4.67·0.1 mL^-1；诃子、矮紫堇、甘青乌头和利巴韦林的最大安全浓度分别为1 mg·mL^-1、1 mg·mL^-1、8 mg·mL^-1和2 μg·mL^-1；3种藏药及利巴韦林在药物和病毒预先作用方式下的最大有效抑制率均高于50%，其中矮紫堇浓度为1 mg·mL^-1时，有效抑制率可达到72.86%，抑制效果极显著高于利巴韦林（P<0.01）；在先加药后加病毒、先加病毒后加药两种作用方式下的最大有效抑制率均小于30%，药物的吸附阻断作用和复制阻断作用并不明显。综上所述，3种藏药对BVDV均有一定的直接杀灭作用。其中，矮紫堇直接杀灭效果最佳且优于利巴韦林，有望被进一步开发成抗病毒药物。

Abstract:

This study was conducted to explore anti-BVDV (bovine viral diarrhea virus) effect of three kinds of Tibetan medicine. BVDV was proliferated by virus propagation technology on bovine kidney cells (MDBK) and the TCID₅₀ was detected. The maximum safe concentration and the CC₅₀ value were measured by CCK8 detection and CPE observation. Antiviral inhibition tests were performed by three different modes, adding virus before adding drugs, adding drugs before adding virus, and adding drugs and virus pre-acted for a while in vitro. The Effective inhibition rate, the EC₅₀ value and the therapeutic index (TI) were calculated by statistical analysis software, which would further clarify the antiviral effection of the extracts. The results showed that TCID₅₀ was 10^-4.67·0.1 mL^-1 by the Reed-Muench method. The maximum safe concentrations of Terminalia chebula, Corydalis hendersonii, Aconitum tanguticum, ribavirin were 1 mg·mL^-1, 1 mg·mL^-1, 8 mg·mL^-1 and 2 μg·mL^-1 respectively. All of the maximum inhibition rates of the drugs were more than 50% in virus-and-drug mode. The most effective inhibitory rate treated with Corydalis hendersonii was 72.86% at the dose of 1 mg·mL^-1, which was significantly higher than that of ribavirin (P<0.01). In addition, all of the maximum inhibitory rates were less than 30% in "virus before drugs" and "virus after drugs" mode, indicating unobvious adsorption block effect and unobvious replication blocking effect. In summary, all three kinds of Tibetan medicine had direct killing effect on BVDV. Corydalis hendersonii had the most significant direct inactivated effect and that was superior to ribavirin, which would expect to be further developed into antiviral drugs.

中图分类号:

王丹阳, 张康, 王旭荣, 王海瑞, 王磊, 张凯, 张景艳, 李建喜, 王学智. 诃子、矮紫堇、甘青乌头提取物对牛病毒性腹泻病毒的体外抑制作用[J]. 畜牧兽医学报, 2018, 49(9): 2036-2043.

WANG Dan-yang, ZHANG Kang, WANG Xu-rong, WANG Hai-rui, WANG Lei, ZHANG Kai, ZHANG Jing-yan, LI Jian-xi, WANG Xue-zhi. Inhibitory Effects of Extracts from Terminalia chebula, Corydalis hendersonii, Aconitum tanguticum on Bovine Viral Diarrhea Virus in vitro[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(9): 2036-2043.

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诃子、矮紫堇、甘青乌头提取物对牛病毒性腹泻病毒的体外抑制作用

Inhibitory Effects of Extracts from Terminalia chebula, Corydalis hendersonii, Aconitum tanguticum on Bovine Viral Diarrhea Virus in vitro

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