Abarema cochliacarpos reduces LPS-induced inflammatory response in murine peritoneal macrophages regulating ROS-MAPK signal pathway
Graphical abstract
Introduction
Inflammation is a complex biological response initiated by several factors that result in cell injury or death contributing to the pathogenesis of many diseases including rheumatoid arthritis, chronic inflammatory bowel disease neurodegenerative disorders, and septic shock syndrome (O’Byrne and Dalgleish, 2001, Kaminska, 2005, Altavilla et al., 2009). It is characterized by the release of cytokines, chemokines, and growth factors and by the transmigration of inflammatory cells, such as monocytes from the blood to the affected tissue, where the differentiation into macrophages and dendritic cells (DCs) occurs, contributing to host defense, tissue remodeling and repair (Gordon and Taylor, 2005, Shi and Pamer, 2011).
Macrophages are major inflammatory and immune effector cells closely related to phagocytic cells that cooperate during the onset, progression and resolution of inflammation (Soehnlein and Lindbom, 2010). These cells are activated by exposure to interferon-γ, pro-inflammatory cytokines, and bacterial lipopolysaccharides (LPS) (Xie et al., 1993, Zhang and Ghosh, 2000). Activated macrophages play pivotal roles in inflammatory diseases via excess production of inflammatory mediators such as nitric oxide (NO) and prostaglandin E2 (PGE2), as well as pro-inflammatory cytokines, to promote inflammatory responses (Vane et al., 1994, Marks-Konczalik et al., 1998). In addition, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) are important enzymes that regulate inflammatory processes (Chang et al., 2011). Moreover, it has been established that gene expression of these pro-inflammatory mediators is well-regulated by mitogen-activated protein kinases (MAPKs) and nuclear transcription factor-kappa B (NF-kB) pathways (Barton and Medzhitov, 2003, Qi and Shelhamer, 2005). On the other hand, the activation of some of these transcription factors may also be influenced by elevated levels of reactive oxygen species (ROS) through a signaling loop (Liu et al., 2005, Yang et al., 2007, Torres and Forman, 2003).
Today, many natural plant products and synthetic compounds have been evaluated for their capacity to modulate inflammatory responses. Among the active constituents of extracts of medicinal plants, polyphenols compounds like flavonoids, are a family of substances whose members have many interesting biological properties, including anti-cancer, anti-microbial, anti-viral, anti-inflammatory, immunomodulatory, and anti-thrombotic activities (Havsteen, 2002).
Abarema cochliacarpos (Gomes) Barneby and Grimes, Fabaceae, is an ornamental tree and native medicinal species in to Brazil. It is popularly known as “barbatimão” or “babatenã”. Wildly it appears in the Atlantic Forest and in the Caatinga (dryland) region of Northeastern Brazil, Cerrado and Campo Rupestre which hosts many different ethnic communities (Albuquerque et al., 2007, Watkinson, 2012). The species was previously listed as Vulnerable (World Conservation Monitoring Center, 1998), however, it is now known to be more widely distributed than previously thought, currently being least concern. However, the Caatinga and Savanna habitat where this species is found is threatened and declining due to agricultural expansion (Watkinson, 2012), being the main threats to this species (The Nature Conservancy, 2010).
The decoction of stem bark of A. cochliacarpos has been used as an herbal remedy in this same region for wound-healing, as an analgesic and anti-inflammatory (Silva et al., 2006, da Silva et al., 2009). Ethnopharmacological studies have demonstrated that A. cochliacarpos is used in traditional medicine for many diseases, including gastrointestinal diseases, through the daily ingestion of this decoction or the tincture, made by placing the bark in white wine or “cachaça” (Silva et al., 2006, Santos, 2008).
Previous studies have shown analgesic and healing effects in skin lesions and antiulcer effects in experimental alcohol gastric ulcer (Santos et al., 2007, Silva et al., 2006, da Silva et al., 2010a). More recently, the butanolic fraction of the methanolic extract was tested on acute and chronic experimental colitis model induced by trinitrobenzene sulfonic acid (TNBS) demonstrating important anti-inflammatory effects (da Silva et al., 2010b, da Silva et al., 2011).
Phytochemical screening of this fraction by electrospray ionization (ESI)/MS has been also performed showing a high content of polyphenols. Its major constituents were catechins, flavonoid-type compounds, and the minor were its dimers and trimers. Catechins refer to monomers of flavanols with similar composition such as catechin, epicatechin, epigallocatechin, epicatechin gallate and epigallocatechin gallate that are particularly abundant in steam bark of the species of Fabaceae family (Santos, 2002). The structure of catechins was confirmed by means of 13C Nuclear Magnetic Resonance analysis characterization, showing that (+)-catechin is the major constituent (da Silva et al., 2010b).
The mechanisms by which catechins achieve their beneficial effects are still not entirely clear; however, there is mounting evidence that they likely work through a combination of both anti-oxidant effect and alteration of intracellular signaling (Fraga and Oteiza, 2011).
With this background the aim of this study was to deepen in the anti-inflammatory effect of butanolic fraction from A. cochliacarpos (BFAC) and its major compound, (+)-catechins, in LPS-stimulated murine peritoneal macrophages.
Section snippets
Plant material and preparation of the extract
Collection and description of the bark of A. cochliacarpos (Gomes) Barneby and Grimes, Fabaceae, as well as preparation of the extract is described by da Silva et al. (2010b)
Sample preparation for UPLC-MS
Extract powder (1 mg) from butanolic fraction was dissolved in MeOH (10 mL). The solution was filtered using a 0.22 μm filter and then diluted with HPLC–MeOH to 100 ppm.
Electrospray ionization-mass spectrometry fingerprint (ESI-MS)
BFAC was diluted in a solution containing 50% (v/v) chromatographic grade methanol (Tedia, Fairfield, OH, USA) and 50% (v/v) deionized water and 0.5% of
Major chemical compounds were identified and quantifying from butanolic fraction by UPLC-MS/MS
Fig. 1 shows the chromatogram obtained from BFAC. Quantifying of the extract components highlighted catechin as the major compound. It was found 32.35 mg of catechin per 1 g of butanolic fraction. The equation of the line is R2=0.9999. These results confirm our previous reported data where (+)-catechin was identified as the major component (da Silva et al., 2010b).
Butanolic fraction from A. cochliacarpos and (+)-catechin in cell viability
In order to evaluate the cell viability in peritoneal macrophages, we assayed the effect of BFAC and (+)-catechin on the growth of
Discussion
A. cochliacarpos has been used to alleviate inflammatory diseases since centuries ago using as crude plant extracts, and previous in vivo studies have discovered an anti-inflammatory activity in different experimental models (da Silva et al., 2010a, da Silva et al., 2010b, da Silva et al., 2011).
BFAC analysis showed that the flavonoid-type compound, (+)-catechin, is its major constituent, followed by catechin and epicatechin dimers. Thus, in this study we explored the ability of the butanolic
Acknowledgments
The research was supported by grants from the Fundación Carolina, Ministerio de Ciencia y tecnología (AGL2008-02475), the Consejería de Innovación Ciencia y Empresa (AGR-6609) and Junta de Andalucía (Spain). The authors gratefully acknowledge the assistance of Center for Technology and Innovation Research, University of Seville (CITIUS).
References (51)
- et al.
Lipid peroxidation is involved in the activation of NF-κB by tumor necrosis factor but interleukin-1 in the human endothelial cell line ECV304
The Journal of Biological Chemistry
(1997) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding
Analytical Biochemistry
(1976)- et al.
Antiinflammatory activities of tormentic acid from suspension cells of Eriobotrya Japonica ex vivo and in vivo
Food Chemistry
(2011) - et al.
MAP kinases in inflammatory bowel disease
Clinica Chimica Acta
(2011) - et al.
Dietary flavonoids: Role of (−)-epicatechin and related procyanidins in cell signaling
Free Radical Biology and Medicine
(2011) - et al.
NF-kappaB activation by reactive oxygen species, fifteen years later
Biochemical Pharmacology
(2006) - et al.
LPS induction of gene expression in human monocytes
Cell Signaling
(2001) - et al.
NF-kappaB and the innate immune response
Current Opinion in Immunology
(2000) The biochemistry and medical significance of the flavonoids
Pharmacology and Therapeutics
(2002)Molecular biology of dysplasia and cancer in inflammatory bowel disease
Gastroenterology Clinics of North America
(2006)
MAPK signalling pathways as molecular targets for anti-inflammatory therapy—from molecular mechanisms to therapeutic benefits
Biochimica et Biophysica Acta
Malabaricone C suppresses lipopolysaccharide-induced inflammatory responses via inhibiting ROS-mediated Akt/IKK/NF-κB signaling in murine macrophages
International Immunopharmacology
Low molecular weight fucoidan from the sporophyll of Undaria pinnatifida suppresses inflammation by promoting the inhibition of mitogen-activated protein kinases and oxidative stress in RAW264.7 cells
Fitoterapia
Cytokine-mediated transcriptional induction of the human inducible nitric oxide synthase gene requires both activator protein 1 and nuclear factor kappaB-binding sites
The Journal of Biological Chemistry
Toll-like receptor 4 signaling regulates cytosolic phospholipase A2 activation and lipid generation in lipopolysaccharide-stimulated macrophages
The Journal of Biological Chemistry
Abarema cochliacarpos: Gastroprotective and ulcer-healing activities
Journal of Ethnopharmacology
Anti-inflammatory intestinal activity of Abarema cochliacarpos (Gomes) Barneby and Grimes in TNBS colitis model
Journal of Ethnopharmacology
Mechanisms of action of green tea catechins, with a focus on ischemia-induced neurodegeneration
The Journal of Nutritional Biochemistry
Protective effect of ebselen on cytotoxicity induced by cholestane-3 beta, 5 alpha, 6 beta-triol in ECV-304 cells
Biochimica et Biophysica Acta
Role of redox potential and reactive oxygen species in stress signaling
Oncogene
Medicinal plants of the caatinga (semi-arid) vegetation of NE Brazil: a quantitative approach
Journal of Ethnopharmacology
Regulation of murine macrophage proinflammatory and anti-inflammatory cytokines by ligands for peroxisome proliferator-activated receptor-gamma: counter-regulatory activity by IFN-gamma
Journal of Leukocyte Biology
Flavocoxid, a dual inhibitor of cyclooxygenase and 5-lipoxygenase, blunts pro-inflammatory phenotype activation in endotoxin-stimulated macrophages
British Journal of Pharmacology
Toll-like receptor signaling pathways
Science
In vivo antioxidant treatment suppresses nuclear factor-κB activation and neutrophilic lung inflammation
The Journal of Immunology
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The two first authors contributed equally.