Effects of <i>Myrcia splendens</i> extract on innate inflammatory response

Pitz, Mayra Alice Corrêa; Benvenutti, Larissa; Paganelli, Camila Jeriane; Reichert, Thaynara; Quintão, Nara Lins Meira; Alberton, Michele Debiasi; Santin, José Roberto; Machado, Isabel Daufenback

doi:10.1590/2175-7860202677007

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Original Paper • Rodriguésia 77 • 2026 • https://doi.org/10.1590/2175-7860202677007 linkcopy

Effects of Myrcia splendens extract on innate inflammatory response

Authorship SCIMAGO INSTITUTIONS RANKINGS

This study investigated the anti-inflammatory activity of the crude hydroalcoholic extract of Myrcia splendens leaves. In vitro, neutrophil chemotaxis and LPS-induced inflammatory mediators were assessed in neutrophils or macrophages treated with M. splendens, salicylic acid, or gallic acid. Inflammation resolution was evaluated via efferocytosis. In vivo effects were tested using the air pouch model. M. splendens impaired macrophage secretion of TNF, IL-1β, IL-6, and NO following LPS stimulation. Neutrophils treated with LPS and incubated with the extract showed reduced migration and NO secretion. Likewise, salicylic and gallic acids decreased NO production in LPS-stimulated neutrophils. The extract enhanced efferocytosis, increased IL-10, and decreased TNF levels. In vivo, similar effects were observed, including reduced neutrophil migration and NO production. These findings suggest that M. splendens modulates neutrophil and macrophage functions, likely due to the presence of salicylic and gallic acids. This highlights its potential as a treatment for inflammatory conditions.

Key words:
inflammation; macrophage; Myrtaceae; neutrophil; resolution of inflammation

Authorship

person Mayra Alice Corrêa Pitz

school Fundação Universidade Regional de Blumenau, Blumenau, SC, Brazil.Fundação Universidade Regional de BlumenauBrazilBlumenau, SC, Brazil Fundação Universidade Regional de Blumenau, Blumenau, SC, Brazil.

0000-0003-1665-6269

person Larissa Benvenutti

school Universidade do Vale do Itajaí, Itajaí, SC, Brazil.Universidade do Vale do ItajaíBrazilItajaí, SC, Brazil Universidade do Vale do Itajaí, Itajaí, SC, Brazil.

0000-0002-9457-9687

person Camila Jeriane Paganelli

0000-0003-4727-4503

person Thaynara Reichert

0009-0005-6013-4419

person Nara Lins Meira Quintão

school Universidade do Vale do Itajaí, Itajaí, SC, Brazil.Universidade do Vale do ItajaíBrazilItajaí, SC, Brazil Universidade do Vale do Itajaí, Itajaí, SC, Brazil.

0000-0001-8305-9339

person Michele Debiasi Alberton

0000-0003-3490-9936

person José Roberto Santin

school Universidade do Vale do Itajaí, Itajaí, SC, Brazil.Universidade do Vale do ItajaíBrazilItajaí, SC, Brazil Universidade do Vale do Itajaí, Itajaí, SC, Brazil.

0000-0002-8805-8818

person Isabel Daufenback Machado 11

0000-0003-0724-5150

person11 Author for correspondence: emailisabelm@furb.br

Area Editor:

Dr. Davyson Moreira

SCIMAGO INSTITUTIONS RANKINGS

Fundação Universidade Regional de Blumenau, Blumenau, SC, Brazil.Fundação Universidade Regional de BlumenauBrazilBlumenau, SC, Brazil Fundação Universidade Regional de Blumenau, Blumenau, SC, Brazil.

Universidade do Vale do Itajaí, Itajaí, SC, Brazil.Universidade do Vale do ItajaíBrazilItajaí, SC, Brazil Universidade do Vale do Itajaí, Itajaí, SC, Brazil.

Figures

Figures (5)

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Figure 1 a-e. Effects of M. splendens on macrophage inflammatory mediator production. The cell viability of murine macrophage strain (RAW.264.7) incubated with M. splendens extract (1, 10 or 100 μg/mL) was accessed by MTT method (a). Cells were stimulated with LPS (5 μg/mL) and then treated with the extract for NO_2- quantification performed by the Griess reaction (b). Quantification of TNF (c), IL-6 (d) and IL-1beta (e) was performed by ELISA according to the manufacturer’s standards in the 1 μg/mL treated cells. Data is expressed as the mean ± S.E.M. of tests performed with cells obtained from 4 independent experiments. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test. **** = p < 0.0001 vs LPS. #### = p < 0.0001 vs basal.

Thumbnail

Figure 2 a-b. Effects of M. splendens on in vitro neutrophils chemotaxis and nitric oxide production. Neutrophils were stimulated with LPS (5 μg/mL) and then treated with the extract for NO_2- quantification, which was performed by the Griess reaction (a). For the chemotaxis assay (b), neutrophil cell suspension was incubated with extract (1, 10 or 100 μg/mL) and placed in front of the fMLP (1 μM) into wells performed in an agarose plate. Cells were counted after 4 h. Quantification was made from the margin of the peripheral perforations towards the chemotactic agent (central perforation). Data is expressed as the mean ± S.E.M. of tests performed with cells obtained from 4 animals. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test. *** = p < 0.001 and **** = p < 0.0001 vs LPS or basal. #### - p < 0.0001 vs basal.

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Figure 3 a-c. Effects of M. splendens extract on in vivo inflammatory cell migration. Air pouches were induced in the dorsal subcutaneous tissue of Swiss mice. The animals were treated orally one hour before the injection of 2 mL of carrageenan. Representative H.E. histological sections of skin biopsies obtained from mice with air pouches (100 and 400x). Black arrows indicate PMN leukocytes. The lavage of the inflammatory infiltrate was collected 4 hours after the injection of carrageenan into the air pouch. The determination of the (a) total number of exudate cells was performed on a Neubauer chamber. (b) The NO level was determined by the Griess reaction method. Values express the mean ± S.E.M. of tests performed with the inflammatory exudate obtained from 6 animals per group. * = p < 0.05 vs control group significantly different from naive group. # = p < 0.001 (One way ANOVA followed by Tukey’s post hoc test).

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Figure 4 a-b. Effects of M. splendens on in vitro efferocytosis. The extract was able to promote the efferocytosis in vitro (a) and reduce the levels of TNF (c) with concomitant increase in IL-10 (b). Data is expressed as a mean ± S.E.M. of cells obtained from four animals. Statistical analysis was performed using One way ANOVA followed by Dunnett’s post hoc test. ** = p < 0.01; *** = p < 0.001 and **** = p < 0.0001 vs basal.

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Figure 5 a-b. Effects of salicylic and gallic acid on in vitro nitric oxide production by neutrophils. Neutrophils were stimulated with LPS (5 μg/mL) and then treated with salicylic acid (a) or gallic acid (b) for NO2- quantification, which was performed by the Griess reaction. Data is expressed as the mean ± S.E.M. of tests performed with cells obtained from four animals. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test. ** = p < 0.01 and **** = p < 0.0001 vs LPS or basal. #### = p < 0.0001 vs basal.

imageFigure 1 a-e. Effects of M. splendens on macrophage inflammatory mediator production. The cell viability of murine macrophage strain (RAW.264.7) incubated with M. splendens extract (1, 10 or 100 μg/mL) was accessed by MTT method (a). Cells were stimulated with LPS (5 μg/mL) and then treated with the extract for NO_2- quantification performed by the Griess reaction (b). Quantification of TNF (c), IL-6 (d) and IL-1beta (e) was performed by ELISA according to the manufacturer’s standards in the 1 μg/mL treated cells. Data is expressed as the mean ± S.E.M. of tests performed with cells obtained from 4 independent experiments. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test. **** = p < 0.0001 vs LPS. #### = p < 0.0001 vs basal. open_in_new

imageFigure 2 a-b. Effects of M. splendens on in vitro neutrophils chemotaxis and nitric oxide production. Neutrophils were stimulated with LPS (5 μg/mL) and then treated with the extract for NO_2- quantification, which was performed by the Griess reaction (a). For the chemotaxis assay (b), neutrophil cell suspension was incubated with extract (1, 10 or 100 μg/mL) and placed in front of the fMLP (1 μM) into wells performed in an agarose plate. Cells were counted after 4 h. Quantification was made from the margin of the peripheral perforations towards the chemotactic agent (central perforation). Data is expressed as the mean ± S.E.M. of tests performed with cells obtained from 4 animals. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test. * = p < 0.001 and ** = p < 0.0001 vs LPS or basal. #### - p < 0.0001 vs basal. open_in_new

imageFigure 3 a-c. Effects of M. splendens extract on in vivo inflammatory cell migration. Air pouches were induced in the dorsal subcutaneous tissue of Swiss mice. The animals were treated orally one hour before the injection of 2 mL of carrageenan. Representative H.E. histological sections of skin biopsies obtained from mice with air pouches (100 and 400x). Black arrows indicate PMN leukocytes. The lavage of the inflammatory infiltrate was collected 4 hours after the injection of carrageenan into the air pouch. The determination of the (a) total number of exudate cells was performed on a Neubauer chamber. (b) The NO level was determined by the Griess reaction method. Values express the mean ± S.E.M. of tests performed with the inflammatory exudate obtained from 6 animals per group. * = p < 0.05 vs control group significantly different from naive group. # = p < 0.001 (One way ANOVA followed by Tukey’s post hoc test). open_in_new

imageFigure 4 a-b. Effects of M. splendens on in vitro efferocytosis. The extract was able to promote the efferocytosis in vitro (a) and reduce the levels of TNF (c) with concomitant increase in IL-10 (b). Data is expressed as a mean ± S.E.M. of cells obtained from four animals. Statistical analysis was performed using One way ANOVA followed by Dunnett’s post hoc test. = p < 0.01; * = p < 0.001 and **** = p < 0.0001 vs basal. open_in_new

imageFigure 5 a-b. Effects of salicylic and gallic acid on in vitro nitric oxide production by neutrophils. Neutrophils were stimulated with LPS (5 μg/mL) and then treated with salicylic acid (a) or gallic acid (b) for NO2- quantification, which was performed by the Griess reaction. Data is expressed as the mean ± S.E.M. of tests performed with cells obtained from four animals. Statistical analysis was performed using one-way ANOVA followed by Dunnett’s post hoc test. = p < 0.01 and ** = p < 0.0001 vs LPS or basal. #### = p < 0.0001 vs basal. open_in_new

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