Immunomodulatory Efficacy of StandardizedAnnona muricata(Graviola) Leaf Extract via Activation of Mitogen-Activated Protein Kinase Pathways in RAW 264.7 Macrophages

Abstract

Annona muricata, commonly known as Graviola, has been utilized as a traditional medicine to treat various human diseases. The aim of this study was to examine the immune-enhancing activity of Graviola leaf extracts in RAW 264.7 macrophage cells. Active ingredients in Graviola leaf extracts (GE) were identified as kaempferol-3-O-rutinoside and quercetin-3-O-rutinoside by LC-MS/MS. When treated with steam or 50% ethanol GE, cell morphology was altered due to initiation of cell differentiation. While the cell viability was not altered by the steam GE, it was reduced by the ethanol GE. Both steam and ethanol GE induced the transcriptional expression of cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-1β, but only the steam extract upregulated inducible nitric oxide synthase (iNOS). In consistence with mRNA expression, the production of TNF-α and nitrite was elevated by both steam and ethanol extracts of Graviola leaves. This is mainly due to activation of mitogen-activated protein (MAP) kinase signaling pathways. These results suggest that Graviola leaves enhance immunity by activation of the MAP kinase pathways. These bioactive properties of Graviola indicate its potential as a health-promoting ingredient to boost the immune system.

Hydroalcoholic Extract and Ethyl Acetate Fraction of Bixa orellana Leaves Decrease the Inflammatory Response to Mycobacterium abscessus Subsp. massiliense

“…The supernatants were discarded, and the formazan crystals were dissolved in 100 µ L dimethyl sulfoxide (DMSO). Absorbance was read at 540 nm using a microplate reader (Thermo Plate), and the results were expressed as a percentage of the maximal value of the positive control and reported as means of three independent assays ± the standard deviation [ 25 , 26 ].…”

“…Cells treated and incubated with 1 μ g/mL LPS ( E. coli 0111 lipopolysaccharide; Sigma-Aldrich, Saint Louis, MO, USA) and 100 μ g/mL IFN- γ (Interferon-gamma; BD Pharmingen) were also used as positive control. After 24 h incubation at 37°C in 5% CO 2 , the supernatants were collected (50 μ L) and incubated with 50 μ L Griess reagent (1% sulfanilamide, 0.1% naphthyl ethylenediamine, and 2.5% phosphoric acid) for 10 min at room temperature in the dark, and the absorbance at 540 nm was compared with that of a standard curve derived from 0−300 µ M NO [ 26 ]. Results were expressed as µ M.…”

Exploring the Leaves of Annona muricata L. as a Source of Potential Anti-inflammatory and Anticancer Agents

“…On the contrary, Kim et al ( 2016 ) reported the immunomostimulatory effect of standardized A. muricata leaf extract via activation of mitogen-activated protein kinase (MAPK) pathway in RAW 264.7 macrophages. The steam and ethanol extracts of A. muricata stimulated the mRNA expression of cytokines, including the pro-inflammatory cytokines, IL-1α and TNF-α, however only the steam extract up regulated inducible nitric oxide synthase (iNOS).…”

Efeito do extrato bruto etanólico da planta Annona muricata L. (Graviola) e suas frações no controle da infecção in vitro e in vivo por Toxoplasma gondii

“…KIM et al, 2016). De forma semelhante, os resultados obtidos na presente investigação mostraram que os níveis de NO são aumentados diante do tratamento com EtOHAm.…”

“…, como também a capacidade em estimular o sistema imune inato, uma vez que o tratamento de macrófagos RAW 264.7 com o extrato de graviola resultou no aumento das concentrações de NO, TNF e IL-1β em seus sobrenadantes(KIM et al, 2016).Entretanto, nossos resultados mostraram que o tratamento com EtOHAm diminuiu os níveis séricos de IFN-γ, TNF e IL-10 dos camundongos infectados com T. gondii. Ainda, os níveis de TNF desses animais foram semelhantes ao grupo não infectado, quando os animais foram tratados com 2,5 ou 5mg de EtOHAm.…”