他莫昔芬对猪伪狂犬病病毒体外增殖的影响

doi:10.16431/j.cnki.1671-7236.2022.11.042

摘要/Abstract

摘要： 【目的】研究他莫昔芬（Tamoxifen）在PK15细胞模型上对猪伪狂犬病病毒（Pseudorabies virus，PRV）感染的抗病毒作用。【方法】以PK15细胞为模型，采用CCK-8细胞计数法检测他莫昔芬对细胞活力的影响；利用ANNEXIN V-FITC/PI凋亡试剂盒检测他莫昔芬对细胞周期和凋亡的影响；利用CytoFLEX流式细胞仪和荧光显微镜检测他莫昔芬处理细胞感染PRV-GFP后病毒增殖的差异；利用实时荧光定量PCR和Western blotting方法分别检测他莫昔芬处理细胞感染PRV-QXX后PRV gB基因mRNA和蛋白表达水平的变化；利用病毒滴度测定法检测他莫昔芬处理细胞感染PRV-QXX后对病毒的抑制情况。【结果】他莫昔芬用药浓度在6 μmol/L时对细胞活力无影响；在6 μmol/L以下时，与空白组相比，他莫昔芬处理组对细胞周期与凋亡没有显著影响（P>0.05）。在同一时间点，他莫昔芬处理组PRV-GFP在PK15细胞中的增殖速度极显著低于对照组（P<0.01）；实时荧光定量PCR结果表明，他莫昔芬处理后极显著抑制了PRV gB基因mRNA在PK15细胞中的表达（P<0.01）；Western blotting结果显示，不同给药浓度同一时间点，他莫昔芬都显著或极显著抑制了PRV gB蛋白的表达（P<0.05；P<0.01）；病毒滴度测定结果表明，在PK15细胞中，同一时间点他莫昔芬处理组的PRV-QXX子代病毒的复制速度显著或极显著低于对照组（P<0.05；P<0.01）。【结论】他莫昔芬能显著抑制PRV在PK15细胞中的增殖。

关键词: 他莫昔芬; 伪狂犬病病毒(PRV); 猪肾上皮细胞; 抗病毒作用

Abstract: 【Objective】 The purpose of this study was to study the antiviral effect of Tamoxifen on the infection of porcine Pseudorabies virus (PRV) in PK15 cell model.【Method】 Taking PK15 cells as the model, the effect of Tamoxifen on cell viability was detected by CCK-8 cell counting method.The effects of Tamoxifen on cell cycle and apoptosis were detected by ANNEXIN V-FITC/PI apoptosis kit.CytoFLEX flow cytometry and fluorescence microscope were used to detect the difference of virus proliferation in Tamoxifen treated cells infected with PRV-GFP.The changes of PRV gB gene mRNA and protein expression in Tamoxifen treated cells infected with PRV-QXX were detected by Real-time quantitative PCR and Western blotting, respectively.Virus titer assay was used to detect the inhibition of Tamoxifen treated cells infected with PRV-QXX.【Result】 Tamoxifen had no effect on cell viability at the concentration of 6 μmol/L.When the concentration of Tamoxifen was below 6 μmol/L, compared with control group, Tamoxifen treatment group had no significant effect on cell cycle and apoptosis(P>0.05).At the same time point, the proliferation rate of PRV-GFP in PK15 cells in Tamoxifen treatment group was extremely significantly lower than that in control group(P<0.01).Real-time quantitative PCR showed that Tamoxifen extremely significantly inhibited the mRNA expression of PRV gB gene in PK15 cells (P<0.01).The results of Western blotting showed that Tamoxifen significantly or extremely significantly inhibited the expression of PRV gB protein in the treatment groups with different concentrations at the same time point (P<0.05 or P<0.01).The results of virus titer showed that the replication rate of PRX-QXX progeny virus in Tamoxifen treatment group was significantly or extremely significantly lower than that in control group at the same time point (P<0.05 or P<0.01).【Conclusion】 Tamoxifen could significantly inhibit the proliferation of PRV in PK15 cells.

Key words: Tamoxifen; Pseudorabies virus (PRV); PK15 cell; antiviral effect

中图分类号:

S859.7

邢嘉友, 樊文杰, 王昱旻, 鲁秀香, 宋月, 褚贝贝, 王江, 王梦迪. 他莫昔芬对猪伪狂犬病病毒体外增殖的影响[J]. 中国畜牧兽医, 2022, 49(11): 4503-4512.

XING Jiayou, FAN Wenjie, WANG Yumin, LU Xiuxiang, SONG Yue, CHU Beibei, WANG Jiang, WANG Mengdi. Effect of Tamoxifen on the Proliferation of Porcine Pseudorabies Virus in vitro[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4503-4512.

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