Nrf2 Activation Protects Against Organic Dust and Hydrogen Sulfide Exposure Induced Epithelial Barrier Loss and K. pneumoniae Invasion.
Nrf2 Activation Protects Against Organic Dust and Hydrogen Sulfide Exposure Induced Epithelial Barrier Loss and K. pneumoniae Invasion.
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DOI：
10.3389/fcimb.2022.848773
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发表时间：
2022
影响因子：
5.7
通讯作者：
中科院分区：
文献类型：
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作者：
农业工人报告由于职业接触有机粉尘(OD)和各种气体而出现各种呼吸道症状。在此之前，我们证明了预先暴露在硫化氢(H_2S)中会改变宿主对OD的反应，并诱导氧化应激。Nrf2是宿主抗氧化反应的主要调节者，暴露于毒物中已知会降低Nrf2的活性。已知过量暴露引起的肺部炎症增加了对继发性微生物感染的易感性。我们验证了一种假设，即反复暴露于OD或H2S会导致Nrf2丢失，上皮细胞失去完整性，并且Nrf2的激活可以拯救这种上皮屏障功能障碍。将原代正常人支气管上皮(NHBE)细胞或小鼠肺精密切片(PCLS)分别用培养液、猪限制设施有机粉尘提取物(ODE)、硫化氢或ODE+H2S处理1~5天。细胞还用载体对照(DMSO)或Nrf2激活剂RTA-408进行预处理。急性暴露于H_2S和ODE+H_2S后，NHBE细胞形态发生改变，细胞存活率下降，Nrf2表达减少，Keap1水平升高。反复暴露于ODE或H2S或ODE+H2S可诱导氧化应激和细胞因子的产生，降低紧密连接蛋白occludin和细胞骨架蛋白Ezrin的表达，破坏上皮完整性，导致肺炎克雷伯菌侵袭增加。RTA-408(Nrf2的药理激活剂)通过降低Keap1水平激活Nrf2，并减轻ODE+H2S诱导的改变，包括逆转PCLS的屏障完整性丧失、炎性细胞因子产生和微生物侵袭，但不能逆转NHBE细胞模型。我们认为，Nrf2的激活对ODE和H_2S具有部分保护作用。
Agriculture workers report various respiratory symptoms owing to occupational exposure to organic dust (OD) and various gases. Previously, we demonstrated that pre-exposure to hydrogen sulfide (H2S) alters the host response to OD and induces oxidative stress. Nrf2 is a master-regulator of host antioxidant response and exposures to toxicants is known to reduce Nrf2 activity. The OD exposure-induced lung inflammation is known to increase susceptibility to a secondary microbial infection. We tested the hypothesis that repeated exposure to OD or H2S leads to loss of Nrf2, loss of epithelial cell integrity and that activation of Nrf2 rescues this epithelial barrier dysfunction. Primary normal human bronchial epithelial (NHBE) cells or mouse precision cut-lung slices (PCLS) were treated with media, swine confinement facility organic dust extract (ODE) or H2S or ODE+H2S for one or five days. Cells were also pretreated with vehicle control (DMSO) or RTA-408, a Nrf2 activator. Acute exposure to H2S and ODE+H2S altered the cell morphology, decreased the viability as per the MTT assay, and reduced the Nrf2 expression as well as increased the keap1 levels in NHBE cells. Repeated exposure to ODE or H2S or ODE+H2S induced oxidative stress and cytokine production, decreased tight junction protein occludin and cytoskeletal protein ezrin expression, disrupted epithelial integrity and resulted in increased Klebsiella pneumoniae invasion. RTA-408 (pharmacological activator of Nrf2) activated Nrf2 by decreasing keap1 levels and reduced ODE+H2S-induced changes including reversing loss of barrier integrity, inflammatory cytokine production and microbial invasion in PCLS but not in NHBE cell model. We conclude that Nrf2 activation has a partial protective function against ODE and H2S.
影响因子：
3.6
作者：
Shrestha D;Bhat SM;Massey N;Santana Maldonado C;Rumbeiha WK;Charavaryamath C
通讯作者：
Charavaryamath C
影响因子：
5.8
作者：
Schneberger D;Pandher U;Thompson B;Kirychuk S
通讯作者：
Kirychuk S
影响因子：
5.8
作者：
Charavaryamath C;Janardhan KS;Townsend HG;Willson P;Singh B
通讯作者：
Singh B