Salbutamol inhibits lipopolysaccharide-induced inflammatory responses in rat peritoneal macrophages

Abstract

-Acute and chronic inflammatory diseases are associated with the induction of inducible nitric oxide synthase (iNOS) and inducible heme oxygenase (HO-1). These inducible enzymes are upregulated in macrophages subjected to inflammatory stimuli and oxidative stress. β 2 -Adrenoceptor (AR) agonists, which function as bronchial dilators, are widely used for the treatment of asthma and chronic obstructive pulmonary disease (COPD). We examined whether salbutamol, a classical β 2 -AR agonist, inhibits the induction of proinflammatory cytokines and stress inducible proteins. Rat macrophages obtained from the abdominal cavity were incubated with lipopolysaccharide (LPS) with or without salbutamol. Induction by LPS of tumor necrosis factor (TNF)-α and interleukin (IL)-6 was significantly inhibited (P < 0.05) by salbutamol treatment. Induction by LPS of iNOS mRNA and protein was also significantly inhibited (P < 0.05) by salbutamol. LPS-mediated increases in HO-1 mRNA and protein were not appreciably affected by salbutamol. One of the anti-inflammatory mechanisms of salbutamol was thus found to be inhibition of induction by LPS of extracellular stimulus-responsive kinase (ERK) 1/2 in macrophages. These findings suggest that salbutamol has the potential for use as an anti-inflammatory agent due to its suppression of LPS-induced TNF-α, and IL-6 and iNOS via ERK pathway without affecting HO-1 expression.Key words: Salbutamol, TNF-α, IL-6, iNOS, LPS Correspondence: Satoru Tanaka (E-mail: satoru_tanaka@pharm.kissei.co.jp) Original ArticleThe Journal of Toxicological Sciences (J. Toxicol. Sci.) Vol.35, No.3, 327-334, 2010 Vol. 35 No. 3 327 enzymes play roles in the pathogenesis of and may therefore be useful as targets in the treatment of asthma and COPD. β 2 -adrenoceptor (AR) agonists such as salbutamol are mainstay bronchodilators in the treatment of asthma and COPD. Malfait et al. (1999) demonstrated that salbutamol exhibited therapeutic effects against rheumatoid arthritis model rats with collagen-induced arthritis. In a clinical study, inhalation of a long-acting β 2 -AR agonist by patients with mild asthma yielded a novel antineutrophilic effect (Jeffery et al., 2002).Based on these findings we examined whether β 2 -AR agonists inhibit proinflammatory cytokine and stressinducible protein production by in vitro cultured abdominal macrophages, in an attempt to determine novel targets in the treatment of inflammatory diseases. MATERIALS AND METHODS MaterialsSalbutamol (hemisulfate, minimum purity 98%) and PD98059, an extracellular stimulus-responsive kinase (ERK) 1/2 inhibitor, were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). LPS from Escherichia coli and ICI118551, a selective β 2 -adrenoceptor antagonist, were purchased from Sigma-Aldrich Inc. (St. Louis, MO, USA). The rat IL-6 Immunoassay KIT and rat TNF-α ELISA KIT were purchased from BioSource International Inc. (Camarillo, CA, USA). Primer sets, including those for iNOS oligo (RA008296), HO-1 oligo (RA008256), and β-actin oligo (RA006...

Epistasis between phenylethanolamine N‐methyltransferase and β2‐adrenergic receptor influences extracellular epinephrine level and associates with the susceptibility to allergic asthma

“…The minor allele “G” of rs11168070 was correlated with increased expression of ADRB2 transcript. As a functional consequence to this, increased ADRB2 signalling in immune cells may lead to suppressed inflammatory response and decreased cytokine secretions in both innate and adaptive T cells, as revealed in prior studies . This supports our subsequent finding that associated allele “G” of rs11168070 with reduced asthma risk.…”

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“…Total RNA was extracted using RNeasy Mini Kit (Qiagen, Valencia, CA, USA). The mRNA levels of tyrosine hydroxylase (TH), heme oxygenase-1 (HO-1), and c-fos in midbrain were examined by quantitative real-time PCR using the StepOne System (Applied Biosystems Inc., Carlsbad, CA, USA), as previously described (Mehta et al, 2009;Tanaka et al, 2010). Primers (TH, HO-1, c-fos and GAP-DH) were purchased from Qiagen.…”

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“…19,20 Briefly, rats (DS and LEW, N¼13, fed a normal diet) were injected i.p. with sterile 3% thioglycolate medium, and 4 days later, the peritoneal exudate corpuscles were collected by lavage with phosphate-buffered saline.…”