Bactericidal antibiotics promote reactive oxygen species formation and inflammation in human sinonasal epithelial cells.
Bactericidal antibiotics promote reactive oxygen species formation and inflammation in human sinonasal epithelial cells.
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DOI：
10.1002/alr.21646
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发表时间：
2016-02
影响因子：
6.4
通讯作者：
Ramanathan M Jr
中科院分区：
文献类型：
--
作者：
Kohanski MA;Tharakan A;Lane AP;Ramanathan M Jr
杀菌抗生素已被证明通过线粒体功能障碍刺激哺乳动物细胞中活性氧（ROS）的形成。这导致氧化性组织损伤，可能对长期使用抗生素产生负面影响。抗生素广泛应用于急慢性鼻窦炎的治疗，但抗生素与鼻窦上皮细胞（SNECs）活性氧（ROS）形成的关系尚未得到证实。在内窥镜鼻窦手术期间从患者中收集人SNEC，并在气液界面处培养生长。用杀菌抗生素阿莫西林和左氧氟沙星以及抑菌抗生素克拉霉素刺激分化的SNEC 24小时。活性氧通过荧光定量。通过LDH分泌定量细胞死亡。采用RT-PCR检测TNF-α和Nrf 2介导的抗氧化基因等炎症标志物的表达。用杀菌抗生素阿莫西林和左氧氟沙星处理培养的SNEC导致ROS产生（p<0.05）和LDH分泌（p<0.05）显著增加。活性氧形成的增加与Nrf-2介导的抗氧化基因表达的增加以及促炎细胞因子TNF- α和IL-1β的表达和产生相关（p<0.05）。用克拉霉素处理的SNEC没有表现出ROS或促炎细胞因子产生的统计学显著增加。在这项研究中，我们证明了用杀菌抗生素治疗培养的人SNEC会导致ROS的形成，并伴随炎症和抗氧化基因表达的增加以及细胞死亡。这表明在鼻窦炎的治疗中长期或不适当地使用抗生素可能导致鼻窦上皮的氧化组织损伤。未来的研究将探讨这种损伤对鼻窦上皮的临床意义。
Bactericidal antibiotics have been shown to stimulate reactive oxygen species (ROS) formation in mammalian cells through mitochondrial dysfunction. This results in oxidative tissue damage that may have negative consequences for long-term antibiotic use. Antibiotics are widely and heavily used in the treatment of acute and chronic sinusitis, however the relationship between antibiotics and ROS formation in sinonasal epithelial cells (SNECs) has not yet been demonstrated. Human SNECs were collected from patients during endoscopic sinus surgery and grown in culture at the air-liquid interface. Differentiated SNECs were stimulated with the bactericidal antibiotics amoxicillin and levofloxacin and the bacteriostatic antibiotic clarithromycin for 24-hours. Reactive oxygen species were quantified via fluorescence. Cell death was quantified by LDH secretion. Expression of inflammatory markers such as TNF-α and Nrf2 mediated antioxidant genes were measured by RT-PCR. Cultured SNECs treated with the bactericidal antibiotics amoxicillin and levofloxacin resulted in a significant increase in production of ROS (p<0.05) and secretion of LDH (p<0.05). The increase in ROS formation correlated with an increase in expression of Nrf-2 mediated antioxidant genes as well as the expression and production of pro-inflammatory cytokine TNF- α, and IL-1β (p<0.05). SNECs treated with clarithromycin did not demonstrate statistically significant increases in ROS or pro-inflammatory cytokine production. In this study, we demonstrate that treatment of cultured human SNECs with bactericidal antibiotics leads to formation of ROS with an associated increase in inflammatory and antioxidant gene expression and cell death. This suggests that long term or inappropriate antibiotic use in the treatment of sinusitis, may result in oxidative tissue damage to the sinonasal epithelium. Future studies will explore the clinical implications of such damage to the sinonasal epithelium.