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. 2016 Nov 23;14(1):179-186.
doi: 10.21010/ajtcam.v14i1.20. eCollection 2017.

IMMUNOSUPPRESIVE EFFECTS OF THE METHANOLIC EXTRACT OF CHRYSOPHYLLUM CAINITO LEAVES ON MACROPHAGE FUNCTIONS

Víctor Ermilo Arana-Argáez  1 , Ivan Chan-Zapata  1 , Jaqueline Canul-Canche  1 , Karla Fernández-Martín  1 , Zhelmy Martín-Quintal  2 , Julio Cesar Torres-Romero  3 , Tania Isolina Coral-Martínez  4 , Julio Cesar Lara-Riegos  3 , Mario Alberto Ramírez-Camacho  5
Affiliations

IMMUNOSUPPRESIVE EFFECTS OF THE METHANOLIC EXTRACT OF CHRYSOPHYLLUM CAINITO LEAVES ON MACROPHAGE FUNCTIONS

Víctor Ermilo Arana-Argáez et al. Afr J Tradit Complement Altern Med. .

Abstract

Background: The aim of this work was to evaluate the immunomodulatory effect of the methanol extract (MeOH) from Chrysophyllum cainito leaves on the MΦs functions.

Material and methods: Peritoneal murine MΦs isolated from Balb/c mice were treated with the MeOH extract and stimulated with LPS. The effect on the phagocytosis was evaluated by flow cytometry assay. The nitric oxide (NO) and hydrogen peroxide (H2O2) production was measured by the Griess reagent and phenol red reaction, respectively. Levels of IL-6 and TNF-α was measured using an ELISA kit. Viability of MΦs and Vero cells was determined by the MTT method.

Results: The MeOH extract of C. cainito leaves inhibited significantly the phagocytosis, and decreased IL-6 and TNF-α production as well as NO and H2O2 released by the MΦs, in a concentration-dependent manner. In addition, MeOH extract of C. cainito showed low cytotoxicity effect against the cells.

Conclusion: These results suggest that MeOH extract of C. cainito leaves has an immunosuppressive effect on murine MΦs, without effects on cell viability. GC-MS chromatogram analysis of MeOH extract showed that lupeol acetate and alpha-amyrin acetate are the principal compounds.

Keywords: Chrysophyllum cainito; Immunomodulation; Macrophages; Phagocytosis; Sapotaceae.

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Figures

Figure 1
Figure 1
Effect of MeOH extract on viability of macrophages. Untreated macrophages were cultured in supplemented DMEM media as negative control or C(-). Macrophages were treated with DMSO 100% as positive control or C(+). b) Effect of MeOH extract on viability of Vero cells. Untreated Vero cells were cultured in supplemented DMEM edia as C(-). Vero cells treated with CDDP (1 tg/mL) were used as C(+). The percentages represent the mean ± SD of three independent experiments (n = 3). Letter “a” indicates significative differences in contrast to C(-) and “b” indicates significant differences in contrast to C(+), according to ANOVA test a dose dependent manner and reflected in the phagocytosis percentages 37.71%, 27.60%, 21.14% and 13.29% for 1 t g/mL, 10 t g/mL, 100 t g/mL and 200 t g/mL respectively. The basal followed by Dunnett post hoc tests (P < 0.05). phagocytic activity found (5.43%) was treated as the negative control or C(-). MΦs stimulated with LPS (1tg/mL) were considered as the positive control C(+), with the highest percentage of phagocytic activity (65.20%).
Figure 2
Figure 2
Effect of MeOH extract on phagocytosis activity of macrophages. Untreated macrophages were co-cultured in supplemented DMEM media and S. cerevisiae yeasts labelled with PI (1 tg/mL) as C(-). Macrophages were cultured in supplemented DMEM media with DMSO 0.1%, treated with LPS (1 tg/mL) and S. cerevisiae yeasts labelled with PI (1 tg/mL) as C(+). The percentages represent the mean ± SD of three independent experiments (n = 3). Letter “a” indicates significative differences in contrast to C(-), according to ANOVA test followed by Dunnett post hoc tests (P < 0.05).
Figure 3
Figure 3
Effect of MeOH extract on NO production of macrophages. Untreated macrophages were cultured in supplemented DMEM media as C(-). Macrophages were cultured in supplemented DMEM media and treated with LPS (1 tg/mL) as C(+). b) Effect of MeOH extract on H2O2 production of macrophages. Untreated macrophages were cultured in supplemented DMEM media as C(-). Macrophages were cultured in supplemented DMEM media and treated with LPS (1 tg/mL) as C(+). The percentages represent the mean ± SD of three independent experiments (n = 3). Letter “a” indicates significative differences in contrast to C(-), according to ANOVA test followed by Dunnett post hoc tests (P < 0.05).
Figure 4
Figure 4
Effect of MeOH extract on IL-6 production of macrophages. Untreated macrophages were cultured in supplemented DMEM media as C(-). Macrophages were cultured in supplemented DMEM media and treated with LPS (1 tg/mL) as C(+). b) Effect of MeOH extract on TNF-α production of macrophages. Untreated macrophages were cultured in supplemented DMEM media as C(-). Macrophages were cultured in supplemented DMEM media and treated with LPS (1 μg/mL) as C(+). The percentages represent the mean ± SD of three independent experiments (n = 3). Letter “a” indicates significative differences in contrast to C(-), according to ANOVA test followed by Dunnett post hoc tests (p < 0.05).
Figure 5
Figure 5
GC chromatographic profile of MeOH extract from C. cainito leaves. (1) alpha-amyrin acetate and (2) lupeol acetate
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