Pseudomonas Quinolone Signal Induces Oxidative Stress and Inhibits Heme Oxygenase-1 Expression in Lung Epithelial Cells
Pseudomonas Quinolone Signal Induces Oxidative Stress and Inhibits Heme Oxygenase-1 Expression in Lung Epithelial Cells
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DOI：
10.1128/iai.00176-17
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发表时间：
2017-09-01
影响因子：
3.1
通讯作者：
Britigan, Bradley E.
中科院分区：
文献类型：
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作者：
Abdalla, Maher Y.;Hoke, Traci;Britigan, Bradley E.
铜绿假单胞菌可导致囊性纤维化患者肺部感染。铜绿假单胞菌喹诺酮信号(PQS)化合物是一种分泌型铜绿假单胞菌毒力因子，参与了铜绿假单胞菌的致病作用。我们能够在感染铜绿假单胞菌的CF患者的痰样本中检测到PQS，但在未感染患者的样本中检测不到PQS。然后，我们通过测定PQS诱导肺上皮细胞(A549和原代正常人支气管上皮细胞[NHBE])细胞和巨噬细胞(J774A)产生活性氧(ROS)的能力，验证了PQS诱导宿主细胞氧化应激的假设。1和THP-1)。用荧光探针(二氯二氢荧光素二醋酸酯、二氢乙锭和MitoSOX红)结合共聚焦显微镜和流式细胞仪检测PQS诱导的ROS产生。PQS诱导肺上皮细胞(A549和NHBE)和巨噬细胞(J774A)产生ROS。1和THP-1细胞)。PQS浓度低至500 ng/ml时，NHBE细胞对PQS敏感。流式细胞仪检测，PQS显著诱导肺上皮细胞早期凋亡(P<；0.05，n=6)。而PQS对J774A细胞的细胞凋亡率无明显影响。1个细胞。血红素氧合酶-1(HO-1)蛋白是一种抗氧化酶，通常由氧化应激诱导。有趣的是，与PQS孵育后，A549和NHBE细胞中HO-1和Nrf2的表达显著降低，而J774A细胞中HO-1的表达显著增加。1细胞(P<；0.05，n=3)，用免疫印迹和密度计法测定。这些PQS对宿主细胞的作用可能在铜绿假单胞菌感染的致病性中起重要作用。
Pseudomonas aeruginosa causes lung infections in patients with cystic fibrosis (CF). The Pseudomonas quinolone signal (PQS) compound is a secreted P. aeruginosa virulence factor that contributes to the pathogenicity of P. aeruginosa. We were able to detect PQS in sputum samples from CF patients infected with P. aeruginosa but not in samples from uninfected patients. We then tested the hypothesis that PQS induces oxidative stress in host cells by determining the ability of PQS to induce the production of reactive oxygen species (ROS) in lung epithelial cells (A549 and primary normal human bronchial epithelial [NHBE]) cells and macrophages (J774A. 1 and THP-1). ROS production induced by PQS was detected with fluorescent probes (dichlorodihydrofluorescein diacetate, dihydroethidium, and MitoSOX Red) in conjunction with confocal microscopy and flow cytometry. PQS induced ROS production in lung epithelial (A549 and NHBE) cells and macrophages (J774A. 1 and THP-1 cells). NHBE cells were sensitive to PQS concentrations as low as 500 ng/ml. PQS significantly induced early apoptosis (P < 0.05, n = 6) in lung epithelial cells, as measured by annexin/propidium iodide detection by flow cytometry. However, no change in apoptosis upon PQS treatment was seen in J774A. 1 cells. Heme oxygenase-1 (HO-1) protein is an antioxidant enzyme usually induced by oxidative stress. Interestingly, incubation with PQS significantly reduced HO-1 and NrF2 expression in A549 and NHBE cells but increased HO-1 expression in J774A. 1 cells (P < 0.05, n = 3), as determined by immunoblotting and densitometry. These PQS effects on host cells could play an important role in the pathogenicity of P. aeruginosa infections.