PCV2通过NF-κB/NLRP3信号通路调控体外培养PAMs分泌IL-1β

doi:10.16431/j.cnki.1671-7236.2016.09.022

《中国畜牧兽医》 ›› 2016, Vol. 43 ›› Issue (9): 2366-2372.doi: 10.16431/j.cnki.1671-7236.2016.09.022

PCV2通过NF-κB/NLRP3信号通路调控体外培养PAMs分泌IL-1β

李海花^1,2, 张蕾^1,2, 杨春蕾^1,2, 赵向华^1,2, 乔家运^1,2, 王文杰^1,2

1. 天津市畜牧兽医研究所, 天津 300381;
2. 天津市畜禽健康养殖技术工程中心, 天津 300381

收稿日期:2016-03-21 出版日期:2016-09-20 发布日期:2016-09-20
通讯作者: 乔家运 E-mail:Qiaojy1979@126.com
作者简介:李海花(1981-),女,河南周口人,博士,助理研究员,研究方向:微生物学与免疫学,E-mail:lihaihuaok@126.com
基金资助:
国家自然科学基金项目（31402087）；天津市农业科学院院长基金（13004）

PCV2 Regulates PAMs Secreting IL-1β through the NF-κB/NLRP3 Signalling Pathway in vitro

LI Hai-hua^1,2, ZHANG Lei^1,2, YANG Chun-lei^1,2, ZHAO Xiang-hua^1,2, QIAO Jia-yun^1,2, WANG Wen-jie^1,2

1. Tianjin Institute of Animal Husbandry and Veterinary Science, Tianjin 300381, China;
2. Tianjin Engineering Research Center for Livestock and Poultry Health Breeding, Tianjin 300381, China

Received:2016-03-21 Online:2016-09-20 Published:2016-09-20

摘要/Abstract

摘要：

为探讨猪圆环病毒2型（porcine circovirus 2，PCV2）诱导猪肺泡巨噬细胞（porcine alveolar macrophages，PAMs）产生白细胞介素-1β（interleukin-1β，IL-1β）的分子机制，试验选取2头PCV2和PRRSV抗原、抗体均为阴性的6周龄普通仔猪，无菌分离PAMs，以体外培养的PAMs为研究对象，采用ELISA方法检测PAMs培养上清液中IL-1β的生成，采用实时荧光定量PCR方法检测PAMs中NLRP3和凋亡相关点样蛋白（apoptosis-associated speck-like protein containing a CARD，ASC）的mRNA表达水平，分别用小干扰RNA（siRNA）方法和核因子-kappa B（NF-κB）抑制试验分析NLRP3和NF-κB对PCV2诱导PAMs产生IL-1β的调控作用。结果显示，PCV2感染PAMs后能够显著或极显著增加IL-1β、NLRP3（1 h除外）和ASC（1、3 h除外）的生成（P<0.05；P<0.01）。siRNA能使58.3%的NLRP3基因沉默，且NLRP3沉默后PCV2诱导PAMs产生IL-1β的水平显著下降（P<0.05）。NF-κB被抑制后PCV2诱导PAMs产生IL-1β的水平也明显下降。结果表明，PCV2通过NF-κB/NLRP3信号通路调控体外培养PAMs分泌IL-1β。

关键词: 猪圆环病毒2型; 猪肺泡巨噬细胞; 白细胞介素-1β 核因子-kappa B; NLRP3

Abstract:

This study investigated the molecular mechanism of interleukin-1β (IL-1β) production in porcine alveolar macrophages (PAMs),which were infected by porcine circovirus 2 (PCV2).Two 6-week-old piglets were selected as experiment animals,which were free of PCV2 and porcine reproductive and respiratory syndrome virus (PRRSV) antibody and antigen.The alveolar macrophages were isolated aseptically and cultured in vitro as this study object.IL-1β in the supernatants from PCV2-infected PAMs was assayed by ELISA.The expressions of NLRP3 and ASC in the PCV2-infected PAMs were detected by Real-time quantitative PCR.The regulatory function of NLRP3 and NF-κB in IL-1β secretion by the PCV2-infected PAMs was analyzed by small interfering RNA (siRNA) and NF-κB inhibitor BAY11-7082,respectively.The results showed that the expressions of IL-1β,NLRP3 (except for 1 h) and ASC (except for 1 and 3 h) in cell culture supernatants or PCV2-infected PAMs were significantly increased(P<0.05;P<0.01).siRNA could lead to 58.3% of NLRP3 gene silencing.IL-1β secretion in the PCV2-infected PAMs,which were companied by NLRP3 gene silencing,was significantly decreased (P<0.05).The IL-1β level in the PCV2-infected PAMs,which were treated with inhibitor BAY11-7082,was also significantly decreased.These results suggested that PCV2 regulated PAMs secreting IL-1β through the NF-κB/NLRP3 signal pathway in vitro.

Key words: PCV2; PAMs; IL-1β; NF-κB; NLRP3

中图分类号:

Q785

李海花, 张蕾, 杨春蕾, 赵向华, 乔家运, 王文杰. PCV2通过NF-κB/NLRP3信号通路调控体外培养PAMs分泌IL-1β[J]. 《中国畜牧兽医》, 2016, 43(9): 2366-2372.

LI Hai-hua, ZHANG Lei, YANG Chun-lei, ZHAO Xiang-hua, QIAO Jia-yun, WANG Wen-jie. PCV2 Regulates PAMs Secreting IL-1β through the NF-κB/NLRP3 Signalling Pathway in vitro[J]. , 2016, 43(9): 2366-2372.

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PCV2通过NF-κB/NLRP3信号通路调控体外培养PAMs分泌IL-1β

PCV2 Regulates PAMs Secreting IL-1β through the NF-κB/NLRP3 Signalling Pathway in vitro

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