Effect of Ambroxol and Beclomethasone on Lipopolysaccharide-Induced Nitrosative Stress in Bronchial Epithelial Cells

Abstract

Background: Nitrosative stress is involved in different airway diseases. Lipopolysaccharide (LPS) induces neutrophil-related cytokine release and nitrosative stress in human bronchial epithelial (BEAS-2B) cells alone or with human polymorphonuclear neutrophils (PMNs). Ambroxol protects against oxidative stress, and beclomethasone dipropionate is an anti-inflammatory drug. Objectives: We evaluated the ability of ambroxol and/or beclomethasone dipropionate to inhibit LPS-induced expression/release of RANTES, IL-8, inducible NO synthase (iNOS), myeloperoxidase (MPO) and 3-nitrotyrosine (3-NT: nitrosative stress biomarker) in BEAS-2B ± PMNs stimulated with LPS (1 μg/ml). Methods: The effect of ambroxol and/or beclomethasone dipropionate on IL-8, RANTES and iNOS levels was assessed by Western blot analysis; IL-8, MPO and 3-NT levels were measured by ELISA. Cell viability was assessed by the trypan blue exclusion test. Results: In BEAS-2B alone, LPS (at 12 h) increased RANTES/iNOS expression and IL-8 levels (p < 0.001). Ambroxol suppressed LPS-induced RANTES expression and IL-8 release (p < 0.001), whilst inhibiting iNOS expression (p < 0.05). Beclomethasone dipropionate had no effect on RANTES but halved iNOS expression and IL-8 release. Coculture of BEAS-2B with PMNs stimulated IL-8, MPO and 3-NT production (p < 0.001), potentiated by LPS (p < 0.001). Ambroxol and beclomethasone dipropionate inhibited LPS-stimulated IL-8, MPO and 3-NT release (p < 0.05). Ambroxol/beclomethasone dipropionate combination potentiated the inhibition of IL-8 and 3-NT production in BEAS-2B with PMNs (p < 0.05 and p < 0.01, respectively). Ambroxol and/or beclomethasone dipropionate inhibited nitrosative stress and the release of neutrophilic inflammatory products in vitro. Conclusion: The additive effect of ambroxol and beclomethasone dipropionate on IL-8 and 3-NT inhibition suggests new therapeutic options in the treatment of neutrophil-related respiratory diseases such as chronic obstructive pulmonary disease and respiratory infections.

Ambroxol for the treatment of fibromyalgia: science or fiction?

“…It can be assumed that a reduction in IL8 is involved: in vitro as well as in vivo, both the release and detectable concentrations of IL8 are reduced by ambroxol, a fact that has been repeatedly shown. 47 , 96 , 201 – 205 This is probably an important finding concerning this “perhaps most important interleukin” in FMS.…”

“…in FMS. Although glucocorticoid tests in 12 female FMS patients showed no reductions in daytime cortisol profile in comparison to 15 controls, they did however show reduced sensitivity of glucocorticoid-receptor function; this was considered a pathophysiologically relevant finding for FMS by Geiss et al 94 In this context, the fact that the anti-inflammatory potency of ambroxol is comparable to dexamethasone 46 , 51 , 95 and beclomethasone 96 without requiring glucocorticoid receptors is not necessarily relevant, but nevertheless worthy of note.…”

“…Apart from obviously enhanced sympathetic activity, FMS patients also have increased IL8 levels in cerebrospinal fluid, 199 , 200 which in principle can be reduced by ambroxol. 96 , 201 – 205 Kadetoff et al 200 interpreted their findings to be a result of FMS symptoms being mediated by sympathetic activity, rather than being dependent on prostaglandin-associated mechanisms, and considered this supportive of the hypothesis of glia-cell activation in response to pain mechanisms. 200 Interestingly, intrathecal administration of ambroxol leads to an antiallodynic effect in an animal model without having an impact on peripheral swelling caused by inflammation.…”

Ion Transport Modulators Differentially Modulate Inflammatory Responses in THP-1-Derived Macrophages

“…Phenytoin and ambroxol HCl are potent inhibitors of voltage-gated sodium channels (Na V s) 1.5 and 1.8, [32,33]. Various authors have previously documented the inhibitory effects of both Na V inhibitors on nitric oxide production [34][35][36]. Phenytoin also inhibits various other macrophage functions including metalloproteinase production, which contributes to gingival hypertrophy in some people on chronic phenytoin therapy [37].…”

Ion transport modulators differentially modulate inflammatory responses in THP-1 derived macrophages

“…Phenytoin and ambroxol HCl are potent inhibitors of voltage-gated sodium channels (Na V s) 1.5 and 1.8 respectively [32,33]. Various authors have previously documented the inhibitory effects of both Na V inhibitors on nitric oxide production [34][35][36]. Phenytoin also inhibits various other macrophage functions including metalloproteinase production, which contributes to gingival hypertrophy in some people on chronic phenytoin therapy [37].…”