Nrf2 protects human alveolar epithelial cells against injury induced by influenza A virus.
Nrf2 protects human alveolar epithelial cells against injury induced by influenza A virus.
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DOI：
10.1186/1465-9921-13-43
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发表时间：
2012-06-06
影响因子：
5.8
通讯作者：
Mason RJ
中科院分区：
文献类型：
--
作者：
Kosmider B;Messier EM;Janssen WJ;Nahreini P;Wang J;Hartshorn KL;Mason RJ
甲型流感病毒（IAV）感染主要针对呼吸道上皮细胞，并产生从轻度上呼吸道感染到严重肺炎的临床结果。最近的研究表明，肺抗氧化防御系统对IAV损伤的重要性。核因子-红细胞2相关因子2（Nrf 2）激活大多数抗氧化基因。从不适合移植的人肺中分离肺泡II型（ATII）细胞和肺泡巨噬细胞（AM），并捐赠用于医学研究。在一些研究中，ATII细胞转分化为肺泡I型样（ATI样）细胞。用A/PR/8/34（PR 8）病毒感染肺泡上皮细胞。我们分析了PR 8病毒的产生、甲型流感病毒核蛋白水平、ROS产生和抗病毒基因的表达。免疫细胞荧光法检测Nrf 2易位，Western blotting法检测Nrf 2、HO-1和caspase 1、3的裂解。我们还通过AM分析了PR 8病毒感染的凋亡ATII细胞的摄取，通过ELISA分析了细胞因子水平，通过TUNEL分析分析了谷胱甘肽水平、坏死和凋亡。此外，我们使用腺病毒Nrf 2（AdNrf 2）或Nrf 2 siRNA分别过表达或敲低Nrf 2来确定Nrf 2的关键重要性。我们发现，IAV诱导氧化应激，细胞毒性和凋亡的ATI样和ATII细胞。我们还发现，AM可以摄取PR 8病毒诱导的凋亡ATII细胞（胞浆细胞增多症），但不能摄取活细胞，而ATII细胞不摄取这些凋亡细胞。PR 8病毒增加ROS的产生，增加Nrf 2、HO-1、Mx 1和OAS 1的表达，增加Nrf 2向细胞核的转运。用siRNA敲低Nrf 2使ATI样细胞和ATII细胞对IAV诱导的损伤敏感，用AdNrf 2过表达Nrf 2保护这些细胞。此外，Nrf 2过表达，随后感染PR 8病毒减少病毒复制，甲型流感病毒核蛋白表达，抗病毒反应和氧化应激。然而，AdNrf 2不增加IFN-λ1（IL-29）水平。我们的研究结果表明，IAV诱导肺泡上皮细胞损伤和Nrf 2保护这些细胞从IAV的细胞病变的影响可能通过增加抗氧化基因的表达。确定在流感感染期间保护细胞免受损伤的途径对于开发新的治疗策略可能特别重要。
Influenza A virus (IAV) infection primarily targets respiratory epithelial cells and produces clinical outcomes ranging from mild upper respiratory infection to severe pneumonia. Recent studies have shown the importance of lung antioxidant defense systems against injury by IAV. Nuclear factor-erythroid 2 related factor 2 (Nrf2) activates the majority of antioxidant genes. Alveolar type II (ATII) cells and alveolar macrophages (AM) were isolated from human lungs not suitable for transplantation and donated for medical research. In some studies ATII cells were transdifferentiated to alveolar type I-like (ATI-like) cells. Alveolar epithelial cells were infected with A/PR/8/34 (PR8) virus. We analyzed PR8 virus production, influenza A nucleoprotein levels, ROS generation and expression of antiviral genes. Immunocytofluorescence was used to determine Nrf2 translocation and western blotting to detect Nrf2, HO-1 and caspase 1 and 3 cleavage. We also analyzed ingestion of PR8 virus infected apoptotic ATII cells by AM, cytokine levels by ELISA, glutathione levels, necrosis and apoptosis by TUNEL assay. Moreover, we determined the critical importance of Nrf2 using adenovirus Nrf2 (AdNrf2) or Nrf2 siRNA to overexpress or knockdown Nrf2, respectively. We found that IAV induced oxidative stress, cytotoxicity and apoptosis in ATI-like and ATII cells. We also found that AM can ingest PR8 virus-induced apoptotic ATII cells (efferocytosis) but not viable cells, whereas ATII cells did not ingest these apoptotic cells. PR8 virus increased ROS production, Nrf2, HO-1, Mx1 and OAS1 expression and Nrf2 translocation to the nucleus. Nrf2 knockdown with siRNA sensitized ATI-like cells and ATII cells to injury induced by IAV and overexpression of Nrf2 with AdNrf2 protected these cells. Furthermore, Nrf2 overexpression followed by infection with PR8 virus decreased virus replication, influenza A nucleoprotein expression, antiviral response and oxidative stress. However, AdNrf2 did not increase IFN-λ1 (IL-29) levels. Our results indicate that IAV induces alveolar epithelial injury and that Nrf2 protects these cells from the cytopathic effects of IAV likely by increasing the expression of antioxidant genes. Identifying the pathways involved in protecting cells from injury during influenza infection may be particularly important for developing new therapeutic strategies.
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影响因子：
3.8
作者：
Mok, Chris K. P.;Lee, Davy C. W.;Lau, Allan S. Y.
通讯作者：
Lau, Allan S. Y.
影响因子：
24.3
作者：
Hodge, S.;Hodge, G.;Reynolds, P. N.
通讯作者：
Reynolds, P. N.
DOI：
10.1165/rcmb.2005-0205oc
发表时间：
2006-02-01
影响因子：
6.4
作者：
Manzer, R;Wang, JR;Mason, RJ
通讯作者：
Mason, RJ
DOI：
10.1164/rccm.200804-535oc
发表时间：
2009-01-15
影响因子：
24.7
作者：
Cho, Hye-Youn;Imani, Farhad;Kleeberger, Steven R.
通讯作者：
Kleeberger, Steven R.
影响因子：
3.7
作者：
Mossel, Eric C.;Wang, Jieru;Jeffers, Scott;Edeen, Karen E.;Wang, Shuanglin;Cosgrove, Gregory P.;Funk, C. Joel;Manzer, Rizwan;Miura, Tanya A.;Pearson, Leonard D.;Holmes, Kathryn V.;Mason, Robert J.
通讯作者：
Mason, Robert J.