艾叶精油体外抑制鸡马立克病病毒增殖的效应分析

doi:10.11843/j.issn.0366-6964.2025.11.045

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (11): 5901-5911.doi: 10.11843/j.issn.0366-6964.2025.11.045

艾叶精油体外抑制鸡马立克病病毒增殖的效应分析

张苑¹(), 李明扬², 梁越², 于向谦², 王妍², 陈吉一², 刘海侠², 连雪²^,*(), 钱莺娟¹^,*()

1. 南京农业大学动物医学院教育部动物健康与食品安全国际合作联合实验室, 南京 210095
2. 江苏农林职业技术学院畜牧兽医学院, 句容 212400

收稿日期:2025-01-20 出版日期:2025-11-23 发布日期:2025-11-27
通讯作者: 连雪,钱莺娟 E-mail:1351040233@qq.com;lianxue@jsafc.edu.cn;yqian@njau.edu.cn
作者简介:张苑(2000-)，女，重庆人，硕士生，主要从事马立克病毒与宿主相互作用研究，E-mail：1351040233@qq.com
基金资助:
国家自然科学基金项目(32202799)；中央高校基本科研业务费专项资金项目(KYCXJC2024005)；江苏省高等学校自然科学研究面上项目(20KJB230011)

Analysis of the Effect of Artemisia argyi Essential Oil on the Inhibition of the Proliferation of Chicken Marek's Disease Virus in vitro

ZHANG Yuan¹(), LI Mingyang², LIANG Yue², YU Xiangqian², WANG Yan², CHEN Jiyi², LIU Haixia², LIAN Xue²^,*(), QIAN Yingjuan¹^,*()

1. MOE Joint International Research Laboratory of Animal Health and Food Safety, College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, China
2. School of Animal Husbandry and Veterinary Medicine, Jiangsu Vocational College of Agriculture and Forestry, Jurong 212400, China

Received:2025-01-20 Online:2025-11-23 Published:2025-11-27
Contact: LIAN Xue, QIAN Yingjuan E-mail:1351040233@qq.com;lianxue@jsafc.edu.cn;yqian@njau.edu.cn

摘要/Abstract

摘要：

本研究旨在探究艾叶精油(Artemisia argyi essential oil，AAEO)对鸡马立克病病毒(Marek’s disease virus，MDV)体外增殖的影响及潜在分子机制。通过CCK-8法测定AAEO对鸡胚成纤维(chicken embryo fibroblast，CEF)细胞的毒性作用，筛选AAEO的最佳作用浓度范围；根据筛选出的最佳作用浓度处理细胞收集样品。通过蚀斑分析和病毒滴度测定，检测AAEO对MDV体外增殖的影响。通过RT-qPCR和Western blot从分子水平上解析AAEO抑制MDV增殖的机制。结果表明，AAEO作用浓度为60~360 μg·mL^-1对CEF无细胞毒作用。AAEO处理组的病毒蚀斑明显减小且变少，病毒滴度下降，病毒在基因水平和蛋白水平的表达上均有下降，说明AAEO可抑制MDV的复制，且具有剂量依赖性。通过对AAEO作用的细胞内信号通路筛选发现，AAEO可下调磷酸化ERK和STAT3蛋白表达水平，下调IL-6 mRNA表达水平。使用ERK抑制剂U0126处理细胞，当细胞内ERK通路被抑制时，病毒复制减少。AAEO可抑制鸡MDV体外增殖，其抗病毒作用可能与AAEO抑制IL-6/STAT3和ERK通路激活有关。艾叶精油具有较好的抗MDV活性，研究结果可为抗疱疹病毒药物的研发提供新的依据。

关键词: 马立克病病毒, 艾叶精油, 鸡胚成纤维细胞, 体外抗病毒

Abstract:

This study aims to investigate the effects of Artemisia argyi essential oil on the in vitro replication of Marek's disease virus and elucidate the underlying molecular mechanisms involved. The cytotoxicity of AAEO on chicken embryo fibroblast (CEF) cells was determined by the CCK-8 method, and the optimal concentration range of AAEO was screened; The cells were treated and samples were collected according to the selected optimal concentration. The effect of AAEO on the proliferation of MDV in vitro was detected by plaque analysis and virus titer assay.The mechanism of AAEO inhibiting MDV proliferation was analyzed at the molecular level by RT-qPCR and Western blot. The results showed that AAEO had no cytotoxic effect on CEF cells in the range of 60-360 μg·mL^-1.In the AAEO treatment group, the virus plaque was significantly reduced and reduced, the virus titer was decreased, and the expression of virus at the gene level and protein level were decreased, indicating that AAEO can inhibit the replication of MDV in a dose-dependent manner. Through the screening of intracellular signaling pathways of AAEO, it was found that AAEO could down regulate the expression levels of phosphorylated ERK and STAT3 proteins, and down regulate the expression levels of IL-6 mRNA. Treatment of cells with the ERK inhibitor U0126 reduced viral replication when the intracellular ERK pathway was inhibited. AAEO effectively inhibits the in vitro proliferation of MDV in chickens. The antiviral mechanism may be associated with its suppression of IL-6/STAT3 signaling pathway and ERK pathway activation. This study demonstrates that AAEO exhibits significant anti-MDV activity, and the findings provide novel evidence for developing therapeutic agents against herpesviruses. The research outcomes offer new insights into the exploration of antiviral drug candidates targeting herpesvirus infections through natural product derivatives.

Key words: Marek's disease virus (MDV), Artemisia argyi essential oil (AAEO), Chicken embryo fibroblast cells, in vitro antiviral

中图分类号:

S859.7

张苑, 李明扬, 梁越, 于向谦, 王妍, 陈吉一, 刘海侠, 连雪, 钱莺娟. 艾叶精油体外抑制鸡马立克病病毒增殖的效应分析[J]. 畜牧兽医学报, 2025, 56(11): 5901-5911.

ZHANG Yuan, LI Mingyang, LIANG Yue, YU Xiangqian, WANG Yan, CHEN Jiyi, LIU Haixia, LIAN Xue, QIAN Yingjuan. Analysis of the Effect of Artemisia argyi Essential Oil on the Inhibition of the Proliferation of Chicken Marek's Disease Virus in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(11): 5901-5911.

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参考文献 29

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抗体 Antibodies	编号 Identifier	来源 Source	宿主 Host	稀释比例 Dilution rate
actin	66009-1-Ig	武汉三鹰生物技术有限公司	鼠	1∶30 000
ERK1/2	11257-1-AP	武汉三鹰生物技术有限公司	兔	1∶1 000
pERK	Cat No. 28733-1-AP	Cell Signaling Technology	兔	1∶500
STAT3	AP0365	bioworld	兔	1∶1 000
p-STAT3(B-7)	#10222	Santa Cruz Biotechnology	鼠	1∶500
VP22	无	以VP22全长为抗原免疫小鼠制备，本实验室保存	鼠	1∶20 000
Goat Anti-Rabbit IgG (H+L) HRP	BS13278	bioworld	兔	1∶10 000
Goat Anti-Mouse IgG (H+L) HRP	BS12478	bioworld	鼠	1∶10 000

引物名称 Primers name	引物对序列(5′→3′) Primer paris suquences
actin	F：GAGACCTTCAACACCCCAGCCATG；R：GCGACGTAGCACAGCTTCTCCTTG
vp22	F：ATCGGATCCATGGGGGATTCTGAAAG；R：ACACTCGAGTTATTCGCTATCACTGC
IL-6	F：CAGGACGAGATGTGCAAG；R：GTAGCACAGAGACTCGACGT

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艾叶精油体外抑制鸡马立克病病毒增殖的效应分析

Analysis of the Effect of Artemisia argyi Essential Oil on the Inhibition of the Proliferation of Chicken Marek's Disease Virus in vitro

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