Pulmonary, gastrointestinal and urogenital pharmacology
Costunolide and Dehydrocostuslactone, two natural sesquiterpene lactones, ameliorate the inflammatory process associated to experimental pleurisy in mice

https://doi.org/10.1016/j.ejphar.2014.02.031Get rights and content

Abstract

The aim of this study was to investigate the effect of costunolide (CS) and dehydrocostuslactone (DCE) a well-known sesquiterpene lactones contained in many plants, in a model of lung injury induced by carrageenan administration in the mice. Injection of carrageenan into the pleural cavity of mice elicited an acute inflammatory response characterized by fluid accumulation in the pleural cavity which contained a large number of polymorphonuclear cells (PMNs) as well as an infiltration of PMNs in lung tissues and increased production of tumour necrosis factor α (TNF-α). All parameters of inflammation were attenuated by CS and DCE (15 mg/kg 10% DMSO i.p.) administered 1 h before carrageenan. Carrageenan induced an up regulation of the intracellular adhesion molecules-1 (ICAM-1) and P-selectin, as well as nitrotyrosine and poly (ADP-ribose) (PAR) as determined by immunohistochemical analysis of lung tissues. The degree of staining for the ICAM-1, P-selectin, nitrotyrosine and PAR was reduced by CS and DCE. Additionally we show that this inflammatory events were associated with NF-κB and STAT3 activation and these sesquiterpenes down-regulated it. Taken together, ours results clearly shown that CS and DCE may offer a novel therapeutic approach for the management of inflammatory diseases.

Introduction

Injection of carrageenan (CAR) into the pleural space leads to local inflammation, infiltration by polymorphonuclear leukocytes (PMNs) and lung injury (Cuzzocrea et al., 1999c). This experimental model of pleurisy has been widely used to investigate the pathophysiology of acute inflammation and also to evaluate the efficacy of drugs in inflammation (Nantel et al., 1999). During the inflammatory response, the high production of reactive oxygen and nitrogen species modulate gene expression as well as apoptosis contributing to the amplification of the inflammatory tissue injury (Fialkow et al., 2007). Recently we and others authors have clearly demonstrated that one mechanism through which reactive oxygen species influences the inflammatory response in the experimental model of lung inflammation is through the activation of the redox-sensitive transcription factor NF-κB (Cuzzocrea et al., 2004), that translocates to the nucleus, where it orchestrates the transcription of a number of pro-inflammatory genes (Peters et al., 2000). Consensus-binding sequences for NF-κB have been identified in the promoter regions of several genes implicated in the pathogenesis of acute and chronic inflammation (Baldwin, 2001).

Another hallmarks of the inflammatory response is the activation of the members of transcription factors, Signal Transducers and Activators of transcription family (STATs), which are induced by cytokines at the early phase of the inflammatory process (Pfitzner et al., 2004). The phosphorylated STATs dimers translocate into the nucleus to regulate expression of target genes (Darnell et al., 1994). Among a number of cytokines released during inflammatory response, interferon (IFN)-γ and interleukin (IL-6) play a pivotal role in regulating the expression of inflammatory genes. IFN-γ exclusively transduces its message by activating cytoplasmic STAT1 whereas the pleiotropic cytokine IL-6 predominantly activates STAT3 and to less extent STAT1. Our previous report (Cuzzocrea et al., 1999b) showed a reduction of carrageenan-elicited lung injury in IL-6-KO mice and a significant attenuation of lung inflammation in the IL-6-WT mice treated with anti-IL-6 antibodies. Furthermore, we have demonstrate that the administration of Hypericum perforatum extract as well as natural molecule like glycyrrhizin protected lung tissues in CAR-treated mice in association with reduced STAT3 activation (Menegazzi et al., 2008a). Recently, we have reported that costunolide (CS) and dehydrocostuslactone (DCE) two sesquiterpene lactones present in a number of plants such as Laurus nobilis, strongly and efficiently inhibit IL-6-induced tyrosine phosphorylation of STAT3 in human leukemic cell line THP-1 (Butturini et al., 2011). Furthermore, these compounds exhibit variety of pharmacological activities including anti-inflammatory and anti-bacterial ones (Robles et al., 1995); pro-apoptotic effect in different human cancer cell line (Hung et al., 2010, Park et al., 2001); inhibitory action on NF-κB activation (Koo et al., 2001); Nfr2 expression (Pae et al., 2007) and MAPK activation (Hsu et al., 2009). Moreover, DCE inhibits the constitutive STAT3 activation (Kuo et al., 2009).

Given that CS and DCE have emerged as a potential anti-inflammatory agent, the present study was designed to explore their possible effect in an experimental model of CAR-induced pleurisy as well as to examine the underlying mechanisms responsible for its effects on this model of acute lung injury.

Section snippets

Animals

Male CD mice (weight 20–25 g; Harlan Nossan, Milan, Italy) were used in these studies. The animals were housed in a controlled environment and provided with standard rodent chow and water. Animal care was in compliance with Italian regulations on the protection of animals used for experimental and other scientific purposes (D.M. 116192) as well as with EEC regulations (O.J. of E.C. L358/1 12/18/1986).

Carrageenan-induced pleurisy

Pleurisy by CAR was induced as previously described (Cuzzocrea et al., 2000). Mice were

Effects of CS and DCE on CAR-induced pleurisy

The injection of carrageenan into the pleural cavity of mice elicited an acute inflammatory response characterized by the accumulation of fluid (exudate) that contained large amounts of PMNs. Exudates volume and neutrophil infiltration in lung tissue were attenuated in a dose-dependent fashion by CS and DCE treatments (Fig. 1A and B, respectively).

The dose of 15 mg/kg was used in the following experiments because it was in the logarithmic dose–response curve of inhibition percent (Fig. 1C and D).

Discussion

Experimental study suggests that pleurisy is a useful model for characterizing and/or screening new anti-inflammatory drugs. The early (4 h) and late (48 h) responses of pleurisy are associated with a marked inflammatory reaction in the airways as a result of the release of distinct mediators and activation of different types of immune cells.

The mouse pleurisy model induced by carrageenan is typically associated with an early response to the phlogistic challenge, where several mediators such as

Conclusion

In summary, our studies led to the conclusion that CS and DCE have potential anti-inflammatory actions in the development of CAR-induced pleurisy and support their therapeutic potential for lung inflammatory pathologies.

Acknowledgements

The authors would like to thank Carmelo La Spada and Giovanni Leotta for their excellent technical assistance during this study, Mrs Caterina Cutrona for secretarial assistance and Miss Valentina Malvagni for editorial assistance with the manuscript.

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