LncRNA 18850对猪流行性腹泻病毒复制的影响

doi:10.11843/j.issn.0366-6964.2025.03.035

畜牧兽医学报 ›› 2025, Vol. 56 ›› Issue (3): 1366-1375.doi: 10.11843/j.issn.0366-6964.2025.03.035

LncRNA 18850对猪流行性腹泻病毒复制的影响

余昕雅¹(), 何海健², 王磊¹, 倪语晨¹, 杜静¹, 周莹珊¹, 董婉玉¹^,*(), 王晓杜¹^,*()

1. 浙江农林大学动物医学院, 浙江省畜禽绿色生态健康养殖应用技术研究重点实验室/动物健康互联网检测技术浙江省工程研究中心/浙江省动物医学与健康管理国际科技合作基地/中澳动物健康大数据分析联合实验室/同一健康和食品安全一带一路国际联合实验室, 杭州 311300
2. 金华职业技术大学农学院, 金华 321017

收稿日期:2024-04-17 出版日期:2025-03-23 发布日期:2025-04-02
通讯作者: 董婉玉,王晓杜 E-mail:1131440070@qq.com;wanyudong@zafu.edu.cn;xdwang@zafu.edu.cn
作者简介:余昕雅(1997-)，女，浙江杭州人，硕士生，主要从事猪传染性腹泻病毒研究，E-mail: 1131440070@qq.com
基金资助:
浙江省自然科学基金探索类项目(LY23C180003)；浙江省自然科学基金基础公益项目(TGN24C180001)；金华市重点科技计划项目(农业类)(2023-2-022)；浙江省领雁计划项目(2023C02022)；农业农村部禽流感等家禽重大疾病防控重点实验室开放课题项目(YDWS202210)；国家级大学生创新创业训练计划项目(202310341032)

Effect of lncRNA 18850 on Porcine Epidemic Diarrhea Virus Replication

YU Xinya¹(), HE Haijian², WANG Lei¹, NI Yuchen¹, DU Jing¹, ZHOU Yingshan¹, DONG Wanyu¹^,*(), WANG Xiaodu¹^,*()

1. Key Laboratory of Applied Biotechnology on Animal Science & Veterinary Medicine of Zhejiang Province, Zhejiang Engineering Research Center for Veterinary Diagnostics & Advanced Technology, Zhejiang International Science and Technology Cooperation Base for Veterinary Medicine and Health Management, China-Australia Joint Laboratory for Animal Health Big Data Analytics, Belt and Road International Joint Laboratory for One Health and Food Safety, College of Veterinary Medicine of Zhejiang A&F University, Hangzhou 311300, China
2. Agriculture College, Jinhua University of Vocational Technology, Jinhua 321017, China

Received:2024-04-17 Online:2025-03-23 Published:2025-04-02
Contact: DONG Wanyu, WANG Xiaodu E-mail:1131440070@qq.com;wanyudong@zafu.edu.cn;xdwang@zafu.edu.cn

摘要/Abstract

摘要：

长链非编码RNA(lncRNA)是一类长度超过200 bp且缺乏蛋白质编码能力的RNA，已有研究表明lncRNA在病毒复制过程中具有重要的生物学作用。本研究分析了猪流行性腹泻病毒(PEDV)感染细胞的lncRNA表达变化及其对PEDV复制的影响。转录组测序PEDV感染Vero-E6细胞24h的样品中lncRNA，构建lncRNA 18850过表达质粒，Western blot、qPCR以及TCID₅₀等方法分析lncRNA 18850过表达对PEDV复制的影响，转录组测序分析lncRNA 18850过表达的Vero-E6细胞差异基因表达，生物信息学分析lncRNA 18850靶向基因及差异蛋白互作关系。结果显示，PEDV感染Vero-E6细胞24和48 h lncRNA 18850的表达量均呈极显著上升(P < 0.01)；lncRNA 18850的过表达可显著促进PEDV的复制(P < 0.05)；LIF、IL11、EPHA2、CCND1、DUSP5、CCN2基因与lncRNA 18850存在靶向关系。PEDV感染可上调Vero-E6细胞内lncRNA 18850的表达，lncRNA 18850可能通过靶向调控多个基因而促进病毒的复制，本研究为深入探析PEDV的复制机制及潜在靶向治疗策略提供了新的视角。

关键词: 长链非编码RNA, 猪流行性腹泻病毒, 转录组测序, Apelin信号通路

Abstract:

Long-stranded non-coding RNAs (lncRNAs) are a class of RNAs longer than 200 bp that lack protein-coding ability. lncRNAs have been shown to play important biological roles in viral replication. This study analyzed changes of lncRNAs in cells infected with porcine epidemic diarrhea virus (PEDV) and their effect on PEDV replication. Transcriptome sequencing was performed on Vero-E6 cells infected with PEDV at different time points. lncRNA 18850 was selected, and its overexpression plasmid was constructed. The effects of lncRNA 18850 overexpression on PEDV replication in Vero-E6 cells were analyzed by Western blot, qPCR and TCID₅₀ methods. Transcriptome sequencing was also used to detect gene changes in Vero-E6 cells overexpressing lncRNA 18850, as well as lncRNA 18850 target genes and differential protein interactions. The results showed that the expression of multiple lncRNAs in PEDV-infected Vero-E6 cells changed significantly, especially the expression of lncRNA 18850, which showed an upward trend at both 24 and 48 hours after viral infection. Overexpression of lncRNA 18850 significantly promoted the replication of PEDV compared with the control group. The genes LIF, IL11, EPHA2, CCND1, DUSP5, and CCN2 in Vero-E6 cells have target relationships with lncRNA 18850. The findings indicate that PEDV infection upregulates the expression of lncRNA 18850 in Vero-E6 cells, and lncRNA 18850 may promote viral replication by regulating multiple target genes, providing new perspectives and insights for a deeper understanding of the replication mechanism of PEDV as well as for exploring the potential therapeutic strategies against it.

Key words: long non-coding RNA, porcine epidemic diarrhea virus, RNA-seq, apelin signaling pathway

中图分类号:

S852.659.6

余昕雅, 何海健, 王磊, 倪语晨, 杜静, 周莹珊, 董婉玉, 王晓杜. LncRNA 18850对猪流行性腹泻病毒复制的影响[J]. 畜牧兽医学报, 2025, 56(3): 1366-1375.

YU Xinya, HE Haijian, WANG Lei, NI Yuchen, DU Jing, ZHOU Yingshan, DONG Wanyu, WANG Xiaodu. Effect of lncRNA 18850 on Porcine Epidemic Diarrhea Virus Replication[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(3): 1366-1375.

图/表 5

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基因 Gene	引物序列(5′→3′) Primer sequences(5′→3′)	长度/bp Length(bp)
lncRNA 18850	GTCCTGCCTCTTCTTTCCATAAG	137
	GAGTTCCAAGTGGCTATCTACAA
CCN2	TGGAAGAGAACATTAAGAAGGGCA	190
	TCAGGGCACTTGAACTCCAC
TFPI2	TGCCAACGTGACTCGCTATT	141
	ATTGGCCTCAACCCTTTTGC
NOTCH3	GGTGGAAGAGCTTATCGCCA	166
	CAGAGGGGTTTCTTCCTTGCT
CXCL8	AAACCACCGGAAGGAACCAT	113
	GGCAAAACTGCACCTTCACA
PTN	TGTTCACCTAACTGCCGTAGCA	101
	TGGTACTGTTGAGCCTGCAT
RRAD	GTATGGTGGGAGACGGATGG	134
	GTCTGTCCACGACCATGAGG
ITGA7	CAAGTATGAAGTCACGGTC	133
	GTCCACATCCAGGTAGAG
GAPDH	GGATGCAGGGATGATGTTC	374
	TGCACCACCAACTGCTTAG

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LncRNA 18850对猪流行性腹泻病毒复制的影响

Effect of lncRNA 18850 on Porcine Epidemic Diarrhea Virus Replication

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