Cytokine gene expression and nitric oxide regulation of Atriplex undulata ethanolic extract in RAW 264.7 macrophage cells

RODRIGUEZ, SILVANA; VIÑA M. D; ANA PAULA MURRAY; LEIRO J.M.

Córdoba

Congreso; 3rd International Meeting on Pharmaceutical Sciences; 2014

Colegio de Farmacéuticos de Córdoba

Atriplex undulata (Moq) D. Dietr. (Chenopodiaceae) commonly known as ?zampa crespa? or ?cachiyuyo?, is an endemic species that is found in Patagonia and the South of Buenos Aires province. The anti-inflammatory activity and the chemical composition of the aerial parts of A. undulata have been evaluated. Ethanolic extract and different successive extracts showed significant anti-inflammatory activity. Moreover, ethanolic and ethyl acetate extracts exhibited better anti-inflammatory activity against interleukin-1β than the other extracts. The ethyl acetate extract was fractionated by column chromatography over silica gel and HPLC, to isolate and purify the active compounds. These compounds were identified by RMN and evaluated for their inhibitory activity of nitric oxide (NO) production in lipopolysaccharide (LPS) stimulated RAW 264.7 macrophage cells. The free and glycosidated flavonoids of ethyl acetate extract have been isolated, such as kaempferol, quercetin, rutin, isorhamnetin and isorhamnetin-diglucopyranoside. The flavonoid fractions and isolated compounds have been evaluated measuring release of interleukins (IL-1β), tumor necrosis factor (TNF-α), expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) by reverse transcriptase-polymerase chain reaction (RT-PCR). In conclusion, this study demonstrated that the isolated components from the ethyl acetate extracts of A. undulata inhibit the production of NO, TNF-α, IL-1β in LPS-stimulated macrophages, in a dose dependent manner. The anti-inflammatory effect was also found to be achieved by the suppression of the production of iNOS and COX-2 inflammatory mediators. The flavonoid glycosides fractions elicited greater anti-inflammatory effect than the isolated compounds. Therefore, this crude extract is a valuable source of anti-inflammatory compounds that has not yet been explored