IFITM3对小反刍兽疫病毒在山羊子宫内膜上皮细胞中增殖的调控效应

doi:10.11843/j.issn.0366-6964.2023.05.040

摘要/Abstract

摘要： 干扰素诱导跨膜蛋白3(interferon-induced transmembrane protein 3,IFITM3)在调控病毒感染过程中发挥重要作用,而其在小反刍兽疫病毒(peste des petits ruminants virus,PPRV)感染中是否发挥作用尚不明确。本研究旨在探究IFITM3对PPRV感染的影响。通过Western blot技术检测PPRV感染对山羊子宫内膜上皮细胞(endometrial epithelial cells,EECs)中IFITM3表达的影响,进一步通过基因过表达及敲降技术探究IFITM3对PPRV复制的影响。结果表明,PPRV感染山羊EECs后,PPRV N蛋白表达水平持续升高,而IFITM3的表达水平较未感染对照组呈先上升后下降趋势。在感染后2 h,IFITM3的表达极显著上调(P<0.001);感染后3~4 h,IFITM3的表达水平最大(P<0.000 1);感染后24 h,IFITM3表达水平下降且与对照组差异不显著。抑制IFITM3表达与对照组相比,PPRV感染后0~6 h,细胞内PPRV的N蛋白水平下降,感染6 h时下降尤为明显(P<0.01);感染后 12~24 h,细胞内PPRV的N蛋白水平变化不明显。过表达IFITM3与对照组相比,感染后6 h,细胞内PPRV的N蛋白水平极显著上升(P<0.001);感染后12~24 h,细胞内PPRV的N蛋白水平较对照组不显著。以上结果表明,PPRV感染山羊EECs早期,通过上调IFITM3表达水平促进PPRV在宿主细胞的复制水平。

关键词: 干扰素诱导跨膜蛋白3, 小反刍兽疫病毒, 山羊EECs, 病毒复制

Abstract: Interferon-induced transmembrane protein 3 (IFITM3) plays an important role in the regulation of viral infection. However, the role of IFITM3 in peste des petits ruminants virus (PPRV) infection remains unclear. This study aimed to investigate the effects of IFITM3 on virus infection in goat endometrial epithelial cells (EECs) infected with PPRV. We performed Western blot assay to detect the kinetics of IFITM3 expression in PPRV-infected EECs. Then, the effect of IFITM3 on PPRV replication was further explored by gene overexpression and knockdown techniques. Our data showed that, compared with mock-infected cells, virus replication increased in PPRV-infected cells in post infection time-dependent manner, which was accompanied with an increased IFITM3 expression followed by decreased expression. The expression of IFITM3 levels increased starting at 2 hours post infection (hpi) (P<0.001), and reached peak at 3-4 hpi (P<0.0001). At 24 hpi, there was no significant difference in IFITM3 expression between PPRV-and mock-infected cells. Moreover, IFITM3 silencing with siRNA significantly inhibit PPRV N expression levels compared to negative control cells at 0-6 hpi, particularly, at 6 hpi. In contrast, over expression of IFITM3 by pcDNA3.1-IFITM3 transfection enhanced PPRV replication levels compared to untransfected cells at 6 hpi (P<0.001). There was no significant difference of virus replication levels in pcDNA3.1-IFITM3 transfected-cells compared to negative control at 12-24 hpi. Together, these results demonstrate that PPRV-induced IFITM3 expression facilitate virus replication during early infection.

Key words: interferon-inducible transmembrane protein 3, peste des petits ruminants virus, Endometriumepithelialcells, virus replication

中图分类号:

S852.659.5

方源, 侯巧弟, 项超辉, 赵红奕, 齐雪峰. IFITM3对小反刍兽疫病毒在山羊子宫内膜上皮细胞中增殖的调控效应[J]. 畜牧兽医学报, 2023, 54(5): 2200-2207.

FANG Yuan, HOU Qiaodi, XIANG Chaohui, ZHAO Hongyi, QI Xuefeng. Regulatory Effects of IFITM3 on Proliferation of Peste des Petits Ruminants Virus (PPRV) in Goat Endometrial Epithelial Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(5): 2200-2207.

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