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. 2017 Jun 20;9(6):635.
doi: 10.3390/nu9060635.

Modulatory Effects of Guarana (Paullinia Cupana) on Adipogenesis

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Free PMC article

Modulatory Effects of Guarana (Paullinia Cupana) on Adipogenesis

Natália da Silva Lima et al. Nutrients. .
Free PMC article

Abstract

Guarana (Paullinia cupana) is a plant originated in Brazil that presents a beneficial effect on body weight control and metabolic alterations. The aim of this study was to evaluate the effects of guarana on genes and miRNAs related to adipogenesis in 3T3L1 cells. The anti-adipogenic effect of guarana was evaluated by Oil Red-O staining. Gene and miRNA expression levels were determined by real time PCR. The Cebpα and β-catenin nuclear translocation were evaluated using immunocytochemistry. Our data indicated that the triglyceride-reducing effect of guarana was dose-dependent from 100 to 300 µg/mL (-12%, -20%, -24% and -40%, respectively, p < 0.0001). An up-regulation of the anti-adipogenic genes Wnt10b, Wnt3a, Wnt1, Gata3 and Dlk1 and a down-regulation of pro-adipogenic genes Cebpα, Pparγ and Creb1 were also observed. Furthermore, guarana repressed mmu-miR-27b-3p, mmu-miR-34b-5p and mmu-miR-760-5p, that contributed for up-regulation of their molecular targets Wnt3a, Wnt1 and Wnt10b. Additionally, cells treated with guarana presented an increase on β-catenin nuclear translocation (p < 0.0018). In summary, our data indicate that guarana has an anti-adipogenic potential due to its ability to modulate miRNAs and genes related to this process. Together our data demonstrate the important role of guarana as a putative therapeutic agent.

Keywords: 3T3L1; Wnt pathway; adipogenesis; guarana (Paullinia cupana); miRNA; obesity.

Conflict of interest statement

The authors declare no conflicts of interest.

Figures

Figure 1
Figure 1
(A) Cell viability evaluated through MTT assay. Control—cells cultivated with DMEM; 50 µg/mL, 100 µg/mL, 150 µg/mL, 200 µg/mL and 300 µg/mL—cells cultivated with DMEM + guarana in different concentrations; (B) Triacylglycerol accumulation in 3T3-L1 cells according to Oil Red O assay. Negative control (C−)—cells cultivated with DMEM; Positive control (C+)—cells cultivated with differentiation medium (DMEM supplemented with 10% fetal bovine serum, 1% of penicillin/streptomycin/glutamine, 0.01% of dexamethasone (1 mM), 0.01% of insulin (100 UI) and 0.1% of IBMX (0.5 mM)) and 50 µg/mL, 100 µg/mL, 150 µg/mL, 200 µg/mL and 300 µg/mL—cells cultivated with differentiation medium + guarana in different concentrations. ### p < 0.0001 compared with C (−) and ** p < 0.001 and *** p < 0.0001 compared to C (+) group; (C) Illustrative images of Oil Red O in 3T3L1 cells (magnification = 200×).
Figure 2
Figure 2
(A) Gene expression of FoxO1, Pparγ, Cebpα, Gata2, Gata3, E2f1, Foxc2, Dlk1, Zfp423 and Creb1 after 96 h of guarana treatment (150 µg/mL) in 3T3L1 cells; (B) Gene expression of Wnt10b, Wnt3a, Wnt1, Sfrp1, Tcf7l2, Lrp5 and Gsk3-β after 96 h of guarana treatment (150 µg/mL) in 3T3L1 cells. White bars correspond to Control group and grey bars correspond to Guarana group. * Significative down-regulation was considered when the fold change was ≤−0.5 and significative up-regulation was considered when the fold change was ≥+2 (* p < 0.05); (C) Immunofluorescence of Cebpα after 96 h of incubation with differentiation medium (Control group) and differentiation medium + guarana 150 µg/mL (Guarana group) in 3T3L1 cells; (D) Density of Cebpα after 96 h of incubation with differentiation medium (Control group—white bar) and differentiation medium + guarana 150 µg/mL in 3T3L1 cells. Density was evaluated using Image J software. *** Guarana group (grey bar) p < 0.0001 compared to Control group (white bar).
Figure 3
Figure 3
(A) Immunofluorescence of β-catenin after 96 h of incubation with differentiation medium (Control group) and differentiation medium + guarana150 µg/mL (Guarana group) in 3T3L1 cells; (B) Density of β-catenin in the nucleus after 96 h of incubation with differentiation medium (Control group—white bar) and differentiation medium + guarana 150 µg/mL in 3T3L1 cells. Density was evaluated using Image J software. ** Guarana group (grey bar) p < 0.001 compared to Control group (white bar); (C) miRNA expression of mmu-miR-27b-3p, mmu-miR-34b-5p and mmu-miR760-5p after 96 h of guarana treatment (150 µg/mL) in 3T3L1 cells. White bars correspond to Control group and grey bars correspond to Guarana group. Error bars reflect SEM. * Significative down-regulation was considered when the fold change was ≤−0.5 and significative up-regulation was considered when the fold change was ≥+2; (D) Gene expression of Wnt1, Wnt3a and Wnt10b in C2C12 cells transfected with mmu-miR-27b-3p, mmu-miR-34b-5p or mmu-miR760-5p mimics. White bars corresponding to Control, striped bars corresponding to C (−) = negative control and grey bars corresponding to mmu-miR-27b-3p, mmu-miR-34b-5p or mmu-miR760-5p mimics.
Figure 4
Figure 4
(A) Gene expression of Wnt1, Wnt3a and Wnt10b in 3T3L1 cells transfected with mmu-miR-27b-3p, mmu-miR-34b-5p or mmu-miR760-5p mimics after 96 h of differentiation. White bars corresponding to Control and grey bars corresponding to 3T3L1 transfected with mmu-miR-27b-3p, mmu-miR-34b-5p or mmu-miR760-5p mimics. * Significative down-regulation was considered when the fold change was ≤−0.5 for microRNA expressions and ≤−0.6 for mimic experiment; (B) Triacylglycerol accumulation in 3T3-L1 cells transfected with mmu-miR-27b-3p, mmu-miR-34b-5p or mmu-miR760-5p mimics according to Oil Red O assay. Control (C+)—cells cultivated with differentiation medium; Guarana (150 µg/mL)—cells cultivated with differentiation medium + guarana (150 µg/mL) during 96 h; 27b-3p, 34b-5p and 760-5p—cells transfected with mmu-miR-27b-3p, mmu-miR-34b-5p or mmu-miR760-5p (respectively) cultivated with differentiation medium. * p < 0.05 and *** p < 0.01 compared to Control; (C) Illustrative images of Oil Red O in 3T3L1 cells Control, Guarana (150 µg/mL) and transfected with mmu-miR-27b-3p, mmu-miR-34b-5p or mmu-miR760-5p mimics (magnification = 200×).
Figure 5
Figure 5
In vitro effects of guarana in gene and microRNAs involved on adipogenesis.

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