Effect of Phosphatidylinositol‐3 Kinase Inhibitors on the Proliferation of Vesicular Stomatitis Virus in Vitro

doi:10.15933/j.cnki.1004-3268.2025.04.014

Journal of Henan Agricultural Sciences ›› 2025, Vol. 54 ›› Issue (4): 136-143.DOI: 10.15933/j.cnki.1004-3268.2025.04.014

• Animal Husbandry and Veterinary Medicine • Previous Articles Next Articles

Effect of Phosphatidylinositol‐3 Kinase Inhibitors on the Proliferation of Vesicular Stomatitis Virus in Vitro

WANG Mengdi¹，FAN Wenjie²，ZHANG Zhen³，ZHAO Ruihong¹，LI Huawei⁴，LIU Kun⁵，ZHENG Ming¹，WANG Yakai¹

（1.College of Food and Bioengineering，Henan University of Animal Husbandry and Economy，Zhengzhou 450046，China；2.College of Veterinary Medicine，Henan Agricultural University，Zhengzhou 450046，China；3.College of Veterinary Medicine，Henan University of Animal Husbandry and Economy，Zhengzhou 450046，China；4.Institute of Animal Product Quality and Safety Technology，Henan University of Animal Husbandry and Economy，Zhengzhou 450046，China；5.Henan Key Laboratory of Innovation and Utilization of Unconventional Feed Resources，Zhengzhou 450046，China）

Received:2024-09-18 Published:2025-04-15 Online:2025-05-20

磷脂酰肌醇-3 激酶抑制剂对水疱性口炎病毒体外增殖的影响

王梦迪¹，樊文杰²，张震³，赵瑞红¹，李华玮⁴，刘昆⁵，郑鸣¹，王亚锴¹

（1.河南牧业经济学院食品与生物工程学院，河南郑州 450046；2.河南农业大学动物医学院，河南郑州 450046；3.河南牧业经济学院动物医药学院，河南郑州 450046；4.河南牧业经济学院畜产品质量安全技术研究院，河南郑州 450046；5.河南省非常规饲料资源创新利用重点实验室，河南郑州 450046）

通讯作者: 王亚锴（1987-），男，河南长垣人，讲师，硕士，主要从事动物生理生化研究。E-mail：yk_wang87@163.com
作者简介:王梦迪（1989-），女，河南新野人，讲师，博士，主要从事动物生理生化研究。E-mail：mengdi.ok@163.com。樊文杰为
同等贡献作者
基金资助:
国家自然科学基金项目（32402849）；河南省重点研发专项（241111110300）；河南省科技攻关项目（252102110035，
222102110158）；河南省教育科学规划一般课题（2024YB0273）；河南牧业经济学院教育教学改革研究与实践项目（2024XJJYXM46）

Abstract

Abstract: The aims are to study the effect of phosphatidylinositol‐3 kinase inhibitor（PI3K‐IN‐1）on the proliferation of vesicular stomatitis virus（VSV）in vitro on PK‐15 cell model，and to reveal the antiviral effect of PI3K‐IN‐1 on VSV in vitro，which will lay a foundation for seeking and developing drugs to prevent and control VSV infection.The safe concentration of PI3K‐IN‐1 at the cellular level was detected by CCK‐8 method；flow cytometry was used to detect the effect of PI3K‐IN‐1 on the percentage of each period in the cell cycle and apoptosis；and flow cytometry and fluorescence microscopy were used to analyze the difference of virus proliferation in PK‐15 cells infected with VSV after PI3K‐IN‐1 treatment；the mRNA expression level of VSV N gene was analyzed by RT‐qPCR；the expression level of GFP protein in VSV‐GFP was analyzed by immunoblotting；the effect of PI3K‐IN‐1 on the titer of VSV progeny virus was detected by virus titer assay；and the inhibitory effect of PI3K‐IN‐1 on the fluorescence intensity of GFP protein in VSV‐GFP was analyzed by confocal microscopy.The results showed that 0—10 μmol/L PI3K‐IN‐1 had no effect on PK‐15 cell viability，percentage of each period in the cell cycle and apoptosis rate. But with the increase of PI3K‐IN‐1 concentration，the expression level of GFP protein in VSV‐GFP decreased. When the concentration of PI3K‐IN‐1 was 10 μmol/L，the inhibitory effect on VSV proliferation was the best. The relative expression level of VSV N gene mRNA in PI3K‐IN‐1 treatment group was significantly or extremely significantly lower than that in DMSO（CK）group and showed a time‐dependent trend.At 12 h and 24 h after VSV‐GFP infection，the proportion of GFP positive fluorescent cells in PI3K‐IN‐1 treatment group was extremely significantly lower than that in CK group，and the progeny virus titer of PI3K‐IN‐1 treatment group was significantly lower than that of CK at 12 h and 24 h after infection，and was extremely significantly lower at 36 h after infection. In summary，PI3K‐IN‐1 can effectively inhibit the proliferation of VSV in PK‐15 cells，which provides a new strategy and method for the research and development of anti‐VSV drugs.

Key words: Vesicular stomatitis virus, Phosphatidylinositol‐3 kinase inhibitor, Antivirus, Proliferation in vitro

摘要： 研究磷脂酰肌醇-3激酶抑制剂（PI3K-IN-1）在PK-15细胞模型上对水疱性口炎病毒（Vesicular stomatitis virus，VSV）体外增殖的影响，揭示PI3K-IN-1在体外细胞中对VSV的抗病毒作用，为寻求和开发防控VSV感染的药物奠定基础。采用CCK-8法检测PI3K-IN-1在细胞水平上的安全使用浓度；采用流式细胞术检测PI3K-IN-1对细胞周期中各时期占比和凋亡率的影响；利用流式细胞术和荧光显微镜分析PI3K-IN-1处理PK-15细胞感染VSV后病毒增殖的差异；利用RT-qPCR分析VSV N 基因的mRNA表达量；利用免疫印迹技术分析VSV-GFP中GFP蛋白的表达量；利用病毒滴度测定试验检测PI3K-IN-1对VSV子代病毒滴度的影响；利用共聚焦显微镜分析PI3K-IN-1对VSV-GFP中GFP蛋白荧光强度的抑制作用。结果表明，0~10 μmol/L PI3K-IN-1对PK-15细胞活力、细胞周期中各时期占比和细胞凋亡率无影响，但随着PI3K-IN-1浓度的增加，VSV-GFP中GFP蛋白的表达量降低，当PI3K-IN-1的浓度为10 μmol/L时，对VSV增殖的抑制效果最好；PI3K-IN-1处理组VSV N 基因mRNA相对表达量显著或极显著低于DMSO（CK）组且呈时间依赖趋势；VSV-GFP感染12 h和24 h时，PI3K-IN-1处理组与CK相比，GFP阳性荧光细胞比例极显著降低；PI3K-IN-1处理组子代病毒滴度与CK相比在感染12、24 h时显著降低，在感染36 h时极显著降低。综上，PI3K-IN-1可有效抑制VSV在PK-15细胞中的增殖，为研究和开发抗VSV药物提供了新的策略与方法。

关键词: 水疱性口炎病毒, 磷脂酰肌醇-3激酶抑制剂, 抗病毒, 体外增殖

CLC Number:

S852.65

WANG Mengdi, FAN Wenjie, ZHANG Zhen, ZHAO Ruihong, LI Huawei, LIU Kun, ZHENG Ming, WANG Yakai. Effect of Phosphatidylinositol‐3 Kinase Inhibitors on the Proliferation of Vesicular Stomatitis Virus in Vitro[J]. Journal of Henan Agricultural Sciences, 2025, 54(4): 136-143.

王梦迪, 樊文杰, 张震, 赵瑞红, 李华玮, 刘昆, 郑鸣, 王亚锴. 磷脂酰肌醇-3 激酶抑制剂对水疱性口炎病毒体外增殖的影响[J]. 河南农业科学, 2025, 54(4): 136-143.

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Effect of Phosphatidylinositol‐3 Kinase Inhibitors on the Proliferation of Vesicular Stomatitis Virus in Vitro

磷脂酰肌醇-3 激酶抑制剂对水疱性口炎病毒体外增殖的影响

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