There is growing evidence that the cannabinoid CB1 receptor antagonist, rimonabant (SR141716) exerts potential anti-proliferative and anti-inflammatory actions. Here, we have assessed the effects of rimonabant in vitro in murine immortalized keratinocytes and in vivo by assaying the topical anti-inflammatory activity.
EXPERIMENTAL APPROACHCell viability and death in a keratinocyte cell line (C5N cells) were measured by Trypan blue exclusion assay and cytotoxicity by sulphorhodamine B test. Cell cycle progression was assessed by flow cytometry and the expression of apoptotic and anti-apoptotic markers, cyclins, pathways of signal transduction and CB1 receptor levels were evaluated by Western blot. The topical anti-inflammatory properties of rimonabant were analysed by inhibition of croton oil-induced ear dermatitis in mice.
KEY RESULTSRimonabant reduced cell viability and induced apoptosis as shown by the enhanced number of cells in the subG0 phase of the cell cycle, the expression of Bax and reduced levels of Bcl-2 and X-inhibitor of apoptosis protein. In addition, reduced levels of phosphorylated serine/threonine protein kinase Akt and nuclear factor-kappa B were detected associated with regulation of total nuclear factor-kappa B and inhibitor of kappa B-a, phosphorylated inhibitor of kappa B-a, cyclins D1, E and A. In croton oil-induced ear dermatitis, rimonabant significantly reduced oedema and leukocyte infiltrate.
CONCLUSIONS AND IMPLICATIONSRimonabant reduced cell viability, inducing cell death in keratinocytes and decreased croton oil-induced ear dermatitis. Our findings suggest a potential application of rimonabant as a topical anti-inflammatory drug. We did not assess the involvement of CB1 receptors in these effects of rimonabant.
AbbreviationsFBS, fetal bovine serum; NSAID, non-steroidal anti-inflammatory drug; TMB, tetramethylbenzidine; TRPV1, vanilloid receptor BJP British Journal of Pharmacology DOI:10.1111DOI:10. /j.1476DOI:10. -5381.2010 84 British Journal of Pharmacology (2011) Introduction Rimonabant (SR141716) [N-piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4methyl pyrazole-3-carboxamide] is a potent and selective cannabinoid CB1 receptor antagonist (receptor nomenclature follows Alexander et al., 2009), inhibiting food intake and exhibiting anti-obesity activity (RinaldiCarmona et al., 1994;1995;Colombo et al., 1998;Simiand et al., 1998;Ravinet Trillou et al., 2003;Carai et al., 2005;Jbilo et al., 2005). It is widely used as a tool to investigate mechanisms by which cannabinoid agonists produce their pharmacological effects and may exert several of its actions by blocking the activation of CB1 receptors, thus modulating the endocannabinoid system, which is tonically activated under certain pathophysiological conditions (Di Marzo and Matias, 2005;Engeli et al., 2005;Matias et al., 2006;Jhaveri et al., 2007;Pertwee, 2009). Anti-tumour properties have been attributed to rimonabant by our group in several cancer cell lines in vitro and in vivo (Sarnataro et al., 2006;Santoro et al., 200...