百里香酚体外抗伪狂犬病毒活性评价及其作用方式  

Evaluation of thymol activity against pseudorabies virus in vitro and its action mode

作　　者：宋天浩 庞莲凤 陈凌霜 邓惠丹[1] 徐志文[1] 朱玲[1] 任志华[1] 邓俊良[1] SONG Tianhao;PANG Lianfeng;CHEN Lingshuang;DENG Huidan;XU Zhiwen;ZHU Ling;REN Zhihua;DENG Junliang(College of Veterinary Medicine,Sichuan Agricultural University,Chengdu 611130,China;Animal Disease Prevention and Control Center,Agricultural and Rural Affairs Bureau of Ganluo County,Liangshan Yi Autonomous Prefecture,Sichuan Province,Ganluo 616850,Sichuan,China)

机构地区：[1]四川农业大学动物医学院,四川成都611130 [2]四川省凉山彝族自治州甘洛县农业农村局动物疫病预防控制中心,四川甘洛616850

出　　处：《浙江农业学报》2023年第1期41-49,共9页Acta Agriculturae Zhejiangensis

基　　金：中央财政农业重大技术协同推广计划(035-2012129217);四川农业大学2019年学科建设双支计划(035-1921993026)。

摘　　要：采用细胞体外培养技术,以BHK-21细胞为研究模型,探讨不同质量浓度的百里香酚在体外抗伪狂犬病毒(PRV)的活性及其作用方式。使用CCK-8法检测百里香酚对BHK-21细胞的最大无害浓度(MNTC),计算得出其半数抑制浓度(IC50)为(212.789±1.652)μg·mL^(-1),对PRV的半数有效浓度(EC50)为(30.710±0.303)μg·mL^(-1),对PRV的治疗指数(TI)为6.929,说明百里香酚属于高效低毒类抗PRV药物。使用CPE观察法检测百里香酚对PRV病毒滴度和病毒一步生长曲线的影响,结果显示,百里香酚能够剂量依赖性显著降低PRV的病毒滴度,并降低PRV在BHK-21中增殖后的毒力。采用荧光定量PCR方法检测不同处理下PRV感染的BHK-21细胞的PRV-gE基因的拷贝量,研究百里香酚体外抗PRV的作用方式,结果显示,百里香酚主要是通过抑制PRV在BHK-21细胞中的增殖来发挥抗病毒活性的,同时能够直接杀灭部分PRV病毒粒子。In this experiment, BHK-21 cells were used as the research model to explore the anti pseudorabies virus(PRV) activity and action mode of thymol at different mass concentrations in vitro. The maximum no-cytotoxic concentration(MNTC) of thymol on BHK-21 cells was detected by CCK-8 method, and the half maximal inhibitory concentration(IC50) was calculated to be(212.789±1.652) μg·mL^(-1). The half effective concentration(EC50) of thymol against PRV was determined as(30.710±0.303) μg·mL^(-1)and the therapeutic index(TI) of thymol was 6.929, which indicated that thymol was a high-efficiency and low-toxicity anti PRV drug. The effects of thymol on virus titer and one-step growth curve of PRV were detected by CPE observation method. The results showed that thymol could significantly reduce the virus titer of PRV and the virulence of PRV after proliferation in BHK-21 cells in a dose-dependent manner. The quantitative PCR was used to detect the gene copies of PRV gE in BHK-21 cells after different treatments to study the action mode of anti PRV effect of thymol in vitro. It was shown that the thymol exerted its antiviral activity mainly by inhibiting the proliferation of PRV in BHK-21 cells, as well as it could directly kill some PRV virus particles.

关 键 词：百里香酚 伪狂犬病毒 体外抗病毒活性 作用方式 

分 类 号：S859[农业科学—临床兽医学]