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| 泽漆水提取物对猪流行性腹泻病毒的体外抑制作用研究 |
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钟睦琪1,2,李凯远2,王雪2,刘甜甜2,3,杨明2,3,郝智慧2*
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| (1.青岛农业大学 化学与药学院, 山东 青岛 266109;2.中国农业大学 动物医学院, 北京 100193;3.新疆农业大学 动物医学院, 乌鲁木齐 830099) |
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| 摘要: |
| 为分析泽漆水提取物对猪流行性腹泻病毒(Porcine epidemic diarrhea virus，PEDV)的体外抑制作用，本研究使用CCK-8法检测泽漆水提取物对非洲绿猴肾细胞(Vero CCL-81)增殖活力的影响，通过间接免疫荧光、蛋白免疫印迹、实时荧光定量PCR方法检测泽漆水提取物对PEDV的抑制作用和时间依赖关系。以广谱抗病毒药利巴韦林(Ribavirin)为阳性对照药物，探究泽漆水提取物对PEDV生命周期各个阶段的影响。结果表明:1)泽漆水提取物浓度≤500 μg/mL时在12、24 和36 h细胞无毒性。2)泽漆水提取物以剂量依赖方式显著降低PEDV-N mRNA的表达量及PEDV 病毒滴度。3)泽漆水提取物可长时间有效抑制PEDV-N蛋白的表达，预防给药对PEDV-N蛋白表达无明显抑制效果。4)泽漆水提取物在病毒复制周期的吸附及复制增殖阶段对PEDV-N蛋白的表达具有显著抑制作用，在入侵阶段对PEDV-N蛋白的表达无明显抑制效果。综上，泽漆水提取物在体外对PEDV具有显著抑制作用，主要作用于PEDV生命周期的吸附和复制增殖阶段。本研究为深入挖掘泽漆抗病毒优势奠定了基础。 |
| 关键词: 泽漆水提取物 猪流行性腹泻病毒 体外抑制 |
| DOI：10.11841/j.issn.1007-4333.2023.10.12 |
| 投稿时间：2023-01-20 |
| 基金项目:中兽药现代化与产业化技术研发(SQ2022YFD1800005) |
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| Inhibitory effect of aqueous extract from Euphorbia helioscopia L. on PEDV in vitro |
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ZHONG Muqi1,2,LI Kaiyuan2,WANG Xue2,LIU Tiantian2,3,YANG Ming2,3,HAO Zhihui2*
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| (1.College of Chemistry and Pharmacy, Qingdao Agricultural University, Qingdao, Shandong 266109, China;2.College of Veterinary Medicine, China Agricultural University, Beijing 100193, China;3.College of Veterinary Medicine, Xinjiang Agricultural University, Urumqi 830099, China) |
| Abstract: |
| In order to analyze the inhibitory effect of aqueous extract from Euphorbia helioscopia L. on porcine epidemic diarrhea virus(PEDV)in vitro, here, the effect of the aqueous extract on the proliferation activity of African green monkey kidney cells(Vero CCL-81)was detected by CCK-8 assay, the inhibition effect and time-response relationship of aqueous extract from Euphorbia helioscopia L. on PEDV were detected by indirect immunofluorescence, western blotting and real-time quantitative PCR(RT-qPCR). Taking the broad-spectrum antiviral drug ribavirin as positive control drug, we explored the effects of the aqueous extract from Euphorbia helioscopia L. on each stage of PEDV life cycle. The results indicated that: 1)When the concentration of aqueous extract from Euphorbia helioscopia L. was no more than 500 μg/mL, there was no cytotoxicity at 12, 24 and 36 h. 2)The PEDV virus titer and its transcription levels of PEDV-N mRNA were significantly decreased in a dose-dependent manner. 3)The extract effectively inhibited PEDV-N protein expression for a long time and the preventive administration of Euphorbia helioscopia L. aqueous extract had no obvious inhibitory effect on the expression of PEDV-N protein. 4)The aqueous extract of Euphorbia helioscopia L. had significant inhibitory effects on the expression of PEDV-N protein during the attachment and replication-proliferation phases of the virus life cycle and had no significant inhibitory effect on the expression of PEDV-N protein during the invasion phase. In conclusion, the aqueous extract of Euphorbia helioscopia L. had a significant inhibitory effect on PEDV in vitro, mainly in the adsorption and replication-proliferation phase of the PEDV life cycle. The study lays the foundation for further exploration of the antiviral advantages of Euphorbia helioscopia L. . |
| Key words: aqueous extract of Euphorbia helioscopia L. porcine epidemic diarrhea virus inhibition in vitro |
