β-雌二醇和孕酮促进非洲猪瘟病毒体外复制

doi:10.11843/j.issn.0366-6964.2022.07.021

畜牧兽医学报 ›› 2022, Vol. 53 ›› Issue (7): 2252-2259.doi: 10.11843/j.issn.0366-6964.2022.07.021

β-雌二醇和孕酮促进非洲猪瘟病毒体外复制

袁兴国^1,2, 申超超², 赵登率^1,2, 杨博², 崔卉梅^1,2, 郝雨², 杨金柯², 陈学辉², 张婷^1,2*, 张克山^1*, 刘霞², 郑海学²

1. 甘肃农业大学生命科学技术学院, 兰州 730070;
2. 中国农业科学院兰州兽医研究所, 兰州大学兽医学院, 家畜疫病病原生物学国家重点实验室, 兰州 730000

收稿日期:2021-10-27 出版日期:2022-07-23 发布日期:2022-07-23
通讯作者: 张克山,主要从事兽医微生物及其分子生物学研究,E-mail:zks009@126.com;刘霞,主要从事家畜基因克隆及其表达调控相关研究,E-mail:413319674@qq.com
作者简介:袁兴国(1998-),男,甘肃白银人,硕士生,主要从事兽医病毒学研究,E-mail:y15390692865@163.com
基金资助:
甘肃省科技重大专项(20ZD7NA006)；国家重点研发计划项目(2021YFD1801300)；中国农业科学院基本科研业务费(Y2020XK09)；国家自然科学基金专项项目(31941002)

β-estradiol and Progesterone Promote Replication of African Swine Fever Virus in vitro

YUAN Xingguo^1,2, SHEN Chaochao², ZHAO Dengshuai^1,2, YANG Bo², CUI Huimei^1,2, HAO Yu², YANG Jinke², CHEN Xuehui², ZHANG Ting^1,2*, ZHANG Keshan^1*, LIU Xia², ZHENG Haixue²

1. College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070;
2. State Key Laboratory of Veterinary Etiological Biology, College of Veterinary Medicine of Lanzhou University, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Science, Lanzhou 730000, China

Received:2021-10-27 Online:2022-07-23 Published:2022-07-23

摘要/Abstract

摘要： 旨在探究雌二醇(β-estradiol，E2)和孕酮(progesterone，P4)对非洲猪瘟病毒(African swine fever virus，ASFV)体外复制的影响。采集空怀猪血清(non-pregnant swine serum，NPSS)和怀孕猪血清(pregnant swine serum，PSS)处理后，用添加胎牛血清(fetal bovine serum，FBS)、NPSS和PSS的培养液分别培养猪肺泡巨噬细胞(porcine alveolar macrophages，PAMs)和骨髓巨噬细胞(bone marrow-derived macrophages，BMDM)并感染ASFV，利用绝对定量PCR和Western blot检测ASFV复制差异；用E2和P4 ELISA试剂盒检测各组血清中E2和P4含量；并在FBS组中单独或者混合添加E2和P4后感染ASFV，检测ASFV复制差异；用NPSS和PSS培养BMDM 24 h后，相对定量检测β-干扰素及下游抗病毒因子转录差异。PSS培养组ASFV复制高于NPSS培养组；PSS中E2和P4的含量均高于NPSS，且FBS中E2和P4的含量很低；在FBS组中单独或者混合添加E2和P4后ASFV复制量增加；PSS培养BMDM后会抑制β-干扰素及下游抗病毒因子的转录。PSS中高含量的E2和P4促进ASFV复制，本研究为解释临床上猪群感染非洲猪瘟(African swine fever，ASF)后母猪首先发病且怀孕母猪病情较重的现象提供一定的科学依据。

关键词: 非洲猪瘟病毒, 雌二醇, 孕酮, 体外, 促进复制

Abstract: This study aimed to investigate the effect of estradiol (E2) and progesterone (P4) on the replication of African swine fever virus (ASFV) in vitro. Non-pregnant swine serum (NPSS) and pregnant swine serum (PSS) were collected and treated. Porcine alveolar macrophages (PAMs) and Bone marrow-derived macrophages (BMDM) were cultivated with Fetal bovine serum (FBS), NPSS and PSS respectively and infected with ASFV. The difference of ASFV replication were detected by absolute quantitative PCR and Western blot. Using E2 and P4 ELISA kits to detect the contents of E2 and P4 in each serum. ASFV was infected with E2 and P4 alone or in combination, and the replication differences of ASFV were detected by absolute quantitative PCR and Western blot. NPSS and PSS treated BMDM for 24 hours, the transcriptional difference of beta-interferon and downstream antiviral factors was detected by relative quantification. The results showed that ASFV replication in PSS group was more effective than NPSS group. The contents of E2 and P4 in PSS were higher than NPSS, but the contents of E2 and P4 in FBS were low levels, and ASFV replication is more effective after adding E2 and P4 alone or mixed in FBS group. And PSS treatment of BMDM can inhibit the transcription of beta-interferon and downstream antiviral factors. High levels of E2 and P4 in PSS promote ASFV replication in vitro. This study provides a scientific basis for explaining the African swine fever (ASF) occurs preferentially in sows and symptoms are more serious in pregnant sows.

Key words: African swine fever virus, estradiol, progesterone, in, vitro, promote replication

中图分类号:

S852.659.6

袁兴国, 申超超, 赵登率, 杨博, 崔卉梅, 郝雨, 杨金柯, 陈学辉, 张婷, 张克山, 刘霞, 郑海学, 刘湘涛. β-雌二醇和孕酮促进非洲猪瘟病毒体外复制[J]. 畜牧兽医学报, 2022, 53(7): 2252-2259.

YUAN Xingguo, SHEN Chaochao, ZHAO Dengshuai, YANG Bo, CUI Huimei, HAO Yu, YANG Jinke, CHEN Xuehui, ZHANG Ting, ZHANG Keshan, LIU Xia, ZHENG Haixue, LIU Xiangtao. β-estradiol and Progesterone Promote Replication of African Swine Fever Virus in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2252-2259.

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β-雌二醇和孕酮促进非洲猪瘟病毒体外复制

β-estradiol and Progesterone Promote Replication of African Swine Fever Virus in vitro

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