Bactericidal antibiotics promote oxidative damage and programmed cell death in sinonasal epithelial cells

Abstract

Background Antibiotics are widely and heavily used in the treatment of chronic sinusitis. Bactericidal antibiotics can stimulate reactive oxygen species (ROS) formation, a pro-inflammatory response and cell death in cultured human sinonasal epithelial cells(SNECs). Sulforaphane is a potent stimulator of the antioxidant Nrf-2 system and a suppressor of inflammation. In this study we utilized sulforaphane to further explore the relationship between levofloxacin treatment, ROS formation and the cell death response. Methods SNECs were collected from patients during endoscopic sinus surgery and grown in culture at the air-liquid interface. Differentiated SNECs were stimulated with levofloxacin with or without sulforaphane pretreatment. Reactive oxygen species were quantified. Apoptosis markers of Caspase-3 activity and DNA fragmentation were quantified. Results Cultured SNECs treated with levofloxacin resulted in a significant increase in activity of the pro-apoptotic Caspase-3 protease (5.9 fold, p = 0.01). The increase in activity was suppressed by pretreatment with sulforaphane (1.9 fold). ROS levels increased with levofloxacin treatment, (range 1.2-1.8 fold) but were not significantly suppressed by pretreatment with sulforaphane (range 1-1.3 fold). Discussion In this study, we demonstrate that treatment of cultured SNECs with levofloxacin leads to an increase in Caspase-3 activity. Sulforaphane pretreatment suppresses the increased apoptotic response possibly through its antioxidant stimulating properties. Our results suggest that levofloxacin treatment stimulates a potent pro-apoptotic possibly through an ROS-dependent mechanism. Future studies will explore if this antibiotic-induced response is harmful to recovery of function in those with sinusitis.

Antibiotics Affect ROS Production and Fibroblast Migration in an In-vitro Model of Sinonasal Wound Healing

“…Whilst many bactericidal antibiotics have been shown to provoke ROS activity in unwounded cells (Kalghatgi et al, 2013;Kohanski et al, 2016Kohanski et al, , 2017, their effect in the post-operative setting, where they are often used empirically to prevent infection, has been hitherto unknown. Our study elucidates the effect of a wide range of commonly used antibiotics on ROS activity in wounded fibroblast and HNEC cell layers, at concentrations that reflect postoperative dosages.…”

“…Apart from their direct antimicrobial effects, many antibiotics also have immunomodulatory functions and have been shown to affect ROS production. Specifically, bactericidal antibiotics have been shown to increase ROS production, whilst bacteriostatic antibiotics do not (Kohanski et al, 2016(Kohanski et al, , 2017. Among the classes of antimicrobials that have been best characterized in the literature are beta-lactams, macrolides, and quinolones (Kohanski et al, 2016).…”

Antibiotic-dependent relationships between nasal microbiome and secreted proteome in chronic rhinosinusitis and nasal polyps

“…Kohanski et al . (Kohanski et al 2017) reported that levofloxacin, a commonly used antibiotic for upper airway infections, simulated reactive oxygen species (ROS) and caspase-3 activity in cultured human sinonasal epithelial cells. Thus, we speculated that antibiotic use could affect the nasal epithelial cells and alter the proteome secreted by these cells.…”

Impact of antibiotics on smell dysfunction

“…Following literature precedence, 7 , 17 penicillins, quinolones and cephalosporins were classified as the primary bactericidal antibiotics. Macrolides, tetracyclines, sulfonamdies and lincosamides were classified as the primary bacteriostatic antibiotics.…”