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| 猪繁殖与呼吸综合征病毒激活 mTORC1/HIF⁃1α通路诱导炎症的作用机制 |
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李晓冰1, 段滇宁1,2, 黄敏1,2, 江潜城1, 杜以梦1, 刘建奎1,3, 邱龙新1,2, 陈洪博1,2
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1.龙岩学院 生命科学学院，福建 龙岩 364012;2.预防兽医学与生物技术福建省高等学校重点实验室，福建 龙岩 364012;3.动物源性人兽共患病防控福建省高校工程研究中心，福建 龙岩 364012
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| 摘要: |
| 【 目的】探究缺氧诱导因子 1α（ HIF-1α）在猪繁殖与呼吸综合征病毒（PRRSV）诱导的猪肺泡巨噬细胞炎症中的作用机制。【 方法】采用蛋白质免疫印迹试验（Western Blot）和实时荧光定量PCR（ Real-time PCR，RT-qPCR），检测感染PRRSV的猪肺脏、肺泡巨噬细胞 HIF-1α mRNA 和蛋白表达水平。使用BAY87-2243或CoCl2处理PRRSV感染的猪肺泡巨噬细胞，检测PRRSV M蛋白和mRNA表达水平，ELISA检测白细胞介素1β（IL-1β）、白细胞介素6（ IL-6）和肿瘤坏死因子α（ TNF-α）的水平。将猪肺泡巨噬细胞转染TSC2 siRNA后，用mTORC1抑制剂 Rapa和mTOR抑制剂KU0063794处理，用Western Blot检测猪肺泡巨噬细胞mTOR对HIF-1α的调控机制。将猪肺泡巨噬细胞过表达mTORC1后，检测PRRSV对HIF-1α和炎症因子水平的影响。【 结果】PRRSV感染可明显提高猪肺脏、肺泡巨噬细胞HIF-1α mRNA和蛋白表达水平。与PRRSV组相比，HIF-1α激活组（CoCl2处理）促进了PRRSV M蛋白和mRNA的表达，IL-1β、IL-6和TNF-α的含量极显著升高（P<0. 01）；HIF-1α 抑制组（BAY87-2243 处理）降低了PRRSV M蛋白和mRNA的表达，IL-1β、IL-6和TNF-α的含量显著或极显著降低（P<0. 01）。PRRSV促进肺泡巨噬细胞中 mTOR 表达，抑制 mTOR 后 HIF-1α 的表达降低，炎症细胞因子水平显著或极显著降低；重新恢复HIF-1α的表达后，炎症细胞因子水平显著升高（P<0. 05）。mTOR主要通过mTORC1上调 HIF-1α 表达；过表达mTORC1可提高PRRSV M和HIF-1α蛋白水平，促进炎症细胞因子表达（P<0. 05）。【 结论】HIF-1α可调控PRRSV诱导的肺泡巨噬细胞炎症反应，其表达受mTORC1信号通路调节。 |
| 关键词: 猪繁殖与呼吸综合征病毒 缺氧诱导因子 1α 哺乳动物雷帕霉素靶蛋白 炎症因子 肺泡巨噬细胞 |
| DOI：10.13207/j.jnwafu.2026.04.001 |
| 分类号: |
| 基金项目:国家自然科学基金项目（32102628）；福建省自然科学基金项目（ 2022J011157）；龙岩学院博士科研启动基金项目（LB2022009） |
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| Mechanism of inflammation induced by porcine reproductive and respiratory syndrome virus via mTORC1/HIF⁃1α signaling pathway activation |
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LI Xiaobing1,2, DUAN Dianning1,2, HUANG Min1, JIANG Qiancheng1, DU Yimeng1, LIU Jiankui1,3, QIU Longxin1,2, CHEN Hongbo1,2
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1.College of Life Science，Longyan University，Longyan，Fujian 364012，China;2.Key Laboratory of Fujian Universities Preventive Veterinary Medicine and Biotechnology，Longyan，Fujian 364012，China;3.Engineering Research Center for the Prevention and Control of Animal Original Zoonosis，Longyan，Fujian 364012，China
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| Abstract: |
| 【Objective】The study aims to explore the mechanism of hypoxia-inducible factor HIF-1α in porcine reproductive and respiratory syndrome virus（PRRSV）-induced inflammation in porcine alveolar macro‐phages.【Method】HIF-1α mRNA and protein expression levels in PRRSV-infected porcine lungs and alveolar macrophages were assessed using Western Blot and real-time quantitative PCR（ RT-qPCR）. PRRSV-infected porcine alveolar macrophages were treated with BAY87-2243 or CoCl2，followed by detection of PRRSV M protein and mRNA expression，and ELISA quantification of interleukin-1β（IL-1β），interleukin-6（IL-6），and tumor necrosis factor alpha（TNF- α）. TSC2 siRNA-transfected porcine alveolar macrophages were treated with Rapa（mTORC1 inhibitor） and KU0063794（mTOR inhibitor），and the regulatory mechanism of mTOR on HIF-1α in porcine alveolar macrophages was detected by Western Blot. After mTORC1 overexpression in porcine alveolar macrophages，the effects of PRRSV on the levels of HIF-1α and inflammatory cytokines were evaluated.【Result】PRRSV infection significantly elevated HIF-1α mRNA and protein levels in porcine lungs
and alveolar macrophages. Compared with the PRRSV-infected group，the HIF-1α activation group（CoCl2treatment） showed increased expression of PRRSV M protein and mRNA and significantly elevated levels of
IL-1β，IL-6，and TNF- α（P<0. 01）. The HIF-1α inhibition group（BAY87-2243 treatment） exhibited decreased expression of PRRSV M protein and mRNA and significantly reduced levels of IL-1β，IL-6，and TNF-α （P<0. 01）. PRRSV was found to enhance mTOR expression in alveolar macrophages，while mTOR inhibition decreased HIF-1α expression and significantly or extremely significantly reduced inflammatory cytokine levels.Upon re-elevating HIF-1α expression，the inflammatory cytokine levels significantly increased （P<0. 05）.mTOR mainly upregulated HIF-1α expression through mTORC1. Overexpression of mTORC1 leads to increased levels of PRRSV M protein and HIF-1α protein，enhancing inflammatory cytokine expression（P<0. 05）.【Conclusion】HIF-1α plays a pivotal role in regulating PRRSV-induced inflammatory responses of al‐veolar macrophages，with its expression modulated by the mTORC1 signaling pathway. |
| Key words: porcine reproductive and respiratory syndrome virus hypoxia inducible factor 1α mammalian target of rapamycin inflammatory factors alveolar macrophage |
