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A bio-guided assay reveals Cymbopogon citratus as source of new anti-inflammatory drugs

AuthorsFrancisco, Vera; García-Rodríguez, Carmen ; Batista, Maria Teresa
Issue Date2012
CitationXLII Reunião Anual da Sociedade Portuguesa de Farmacologia (2012)
Abstract[Introduction]: Inflammation is associated with several pathophysiological conditions, but the current anti-inflammatory drugs are not totally safe and effective. Therefore, enforcements to develop new and safe anti-inflammatory agents are urgent, being the signal transduction pathways and mediators involved in inflammation potential therapeutic targets. For that reason, inhibition of nuclear factor (NF)-κB, nitric oxide (NO) and cytokines has been pointed as an anti-inflammatory strategy. Additionally, in the last decades, natural products have been an important source of new drugs. Thus, the aim of the present study is to explore, through an in vitro bio-guided assay, the anti-inflammatory activity and the molecular mechanism of Cymbopogon citratus (Cy), a plant used in traditional medicine to treat inflammatory conditions. [Materials and Methods]: A lipid- and essential oil-free infusion of Cy leaves was prepared (Cy extract) and fractionated by column chromatography. Anti-inflammatory properties of Cy extract (1.115 mg/ml) and its polyphenol-rich fractions (PFs), namely phenolic acids (530 μg/ml), flavonoids (97.5 μg/ml) and tannins (78 μg/ml), were investigated using lipopolysaccharide (LPS)-stimulated Raw 264.7 and human macrophages as in vitro model. NO production was evaluated by Griess reaction and inducible NO synthase (iNOS) expression by Western blot. The NF-κB activation was analyzed by Western blot and reporter assay, while the activity of proteasome, a complex which controls the NF-κB activation, was analyzed using a fluorogenic peptide. Cytokine expression was studied by real-time polymerase chain reaction (PCR) and the results analyzed by Pfaffl method. One-way ANOVA followed by Dunnett’s test was applied to compare the effect of different treatments on LPS-stimulated cells. [Results]: We found that Cy maintained the phosphorylation of the inhibitory protein κB (IκB) and inhibited IκB degradation, p65 translocation to the nucleus and NF-κB transcriptional activity. Moreover, the data showed, for the first time, the inhibition of proteasome activity by Cy extract. We also demonstrated anti-inflammatory properties of Cy through inhibition of pro-inflammatory cytokine expression. In turn, PFs inhibited NF-κB activation and cytokine expression, indicating that polyphenols are partially responsible for the anti-inflammatory activity of Cy. These results were confirmed in LPS-stimulated human macrophages. [Conclusions]: Our work supports the use of Cymbopogon citratus infusion in traditional medicine and better elucidates its anti-inflammatory mechanism of action, thus revealing the pharmacological importance of this herb, in particular its polyphenols, as a natural source of new anti-inflammatory drugs.
DescriptionResumen del trabajo presentado a la XLII Reunião Anual da Sociedade Portuguesa de Farmacologia celebrada en Lisboa (Portugal) del 1 al 3 de febrero de 2012.-- et al.
Appears in Collections:(IBGM) Comunicaciones congresos
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