MiR-222对山羊副流感病毒3型复制的影响

doi:10.11843/j.issn.0366-6964.2018.12.015

畜牧兽医学报 ›› 2018, Vol. 49 ›› Issue (12): 2664-2671.doi: 10.11843/j.issn.0366-6964.2018.12.015

MiR-222对山羊副流感病毒3型复制的影响

钟纯燕^1,3, 李基棕^1,2*, 毛立¹, 李文良¹, 郝飞¹, 孙敏¹, 刘茂军¹, 主性³, 嵇辛勤³, 肖芳^1,3, 杨蕾蕾¹, 张纹纹¹

1. 江苏省农业科学院兽医研究所, 农业部兽用生物制品工程技术重点实验室, 南京 210014;
2. 临沂大学药学院, 临沂 276000;
3. 贵州大学动物科学学院, 贵阳 550025

收稿日期:2018-05-07 出版日期:2018-12-23 发布日期:2018-12-23
通讯作者: 李基棕,E-mail:lijizong22@sina.com
作者简介:钟纯燕(1992-),女,贵州兴义人,硕士生,主要从事动物传染病防治和诊断技术研究,Tel:025-84391152,E-mail:1462651179@qq.com
基金资助:
国家自然科学基金（31702272）；江苏省自然科学基金（BK2017059）；山东省自然科学基金（ZR2016CP08）；“十三五”国家重点研发计划项目（2016YFD0500908）

The Molecular Mechanism Study of miR-222 Inhibit Caprine Parainfluenza Virus Type 3 Replication

ZHONG Chun-yan^1,3, LI Ji-zong^1,2*, MAO Li¹, LI Wen-liang¹, HAO Fei¹, SUN Min¹, LIU Mao-jun¹, ZHU Xing³, JI Xin-qin³, XIAO Fang^1,3, YANG Lei-lei¹, ZHANG Wen-wen¹

1. Key Laboratory of Veterinary Biological Engineering and Technology of Ministry of Agriculture, Institute of Veterinary Medicine, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China;
2. School of Pharmacy, Linyi University, Linyi 276000, China;
3. College of Animal Science of Guizhou University, Guiyang 550025, China

Received:2018-05-07 Online:2018-12-23 Published:2018-12-23

摘要/Abstract

摘要：

为探讨miR-222对山羊副流感病毒3型（CPIV3）复制的影响及其机制。本研究将miR-222模拟体转染MDBK细胞，接种CPIV3病毒液，收集上清和细胞，测定病毒效价和干扰素（IFN）含量；通过TargetScan和miRanda等生物信息学软件预测miR-222调控的靶基因，用双荧光素酶报告系统验证其靶基因，RNA干扰技术研究靶基因对CPIV3复制的影响。结果显示，miR-222模拟体能抑制CPIV3复制，转染miR-222组IFN的表达水平较高，且病毒复制效价（10^2.5 TCID₅₀·0.1 mL^-1）显著低于miRNA阴性对照组（10^3.3 TCID₅₀·0.1 mL^-1）；双荧光素酶报告系统验证和qPCR检测得出，miR-222可靶向结合IRF2的3'-UTR，并降解IRF2的mRNA和抑制IRF2蛋白表达；敲除IRF2的MDBK细胞感染CPIV3后，病毒复制效价（10^2.0 TCID₅₀·0.1 mL^-1）低于正常MDBK细胞组（10^3.2 TCID₅₀·0.1 mL^-1）。本研究表明，miR-222通过靶向降解IRF2 mRNA提高IFN表达水平来抑制CPIV3复制，为基于miR-222研制潜在抗病毒药物提供基础资料。

Abstract:

To explore the function of miR-222 on effecting the replication of caprine parainfluenza virus type 3 (CPIV3). The miR-222 mimics were synthesized and transfected into the MDBK cells, then infected with CPIV3. Twenty-four hours after infection, the cell cultures were harvested to determine the expression of CPIV3 genome and IFN mRNA. The molecular mechanism of effecting CPIV3 replication was explored using bioinformatics software, reporter gene system, RT-qPCR, Western blot and RNAi analysis. The results showed that the overexpression of miR-222 promote interferon (IFN) production to inhibit CPIV3 replication. Prediction the targets of miR-222 discovered that interferon regulatory factor 2 (IRF2) has a putative miR-222 target site and miR-222 mimics markedly decreased the luciferase level. Furthermore, transfection with miR-222 mimics decreased IRF2 expression in MDBK cells at both the mRNA and protein levels. Knockdown of miR-222 target inhibited CPIV3 replication. These findings indicate that overexpression of miR-222 promote the host antiviral innate immune response by targeting IRF2.

中图分类号:

S852.659.5

钟纯燕, 李基棕, 毛立, 李文良, 郝飞, 孙敏, 刘茂军, 主性, 嵇辛勤, 肖芳, 杨蕾蕾, 张纹纹. MiR-222对山羊副流感病毒3型复制的影响[J]. 畜牧兽医学报, 2018, 49(12): 2664-2671.

ZHONG Chun-yan, LI Ji-zong, MAO Li, LI Wen-liang, HAO Fei, SUN Min, LIU Mao-jun, ZHU Xing, JI Xin-qin, XIAO Fang, YANG Lei-lei, ZHANG Wen-wen. The Molecular Mechanism Study of miR-222 Inhibit Caprine Parainfluenza Virus Type 3 Replication[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(12): 2664-2671.

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MiR-222对山羊副流感病毒3型复制的影响

The Molecular Mechanism Study of miR-222 Inhibit Caprine Parainfluenza Virus Type 3 Replication

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