Keywords alpha, beta-Amyrin; 3T3-L1 cells; Adipocyte differentiation; PPARgamma; C/EBPalpha; GLUT4; Anti-adipogenecity Highlights * The triterpenoid, alpha, beta-Amyrin manifests anti-adipogenicity in 3T3-L1 cells. * alpha, beta-Amyrin inhibits preadipocyte differentiation. * alpha,beta-Amyrin suppresses adipogenic transcription factors, PPARgamma and C/EBPalpha. * alpha,beta-Amyrin stimulates the glucose transporter GLUT4 and promotes AMPK phosphorylation. Previous studies have reported the anti-obesity effects of alpha, beta-Amyrin in high fat-fed mice. This study aimed to evaluate whether alpha, beta-Amyrin has an anti-adipogenic effect in 3T3-L1 murine adipocytes and to explore the possible underlying mechanisms. 3T3-L1 pre-adipocytes were differentiated in a medium containing insulin, dexamethasone, and 1-methyl-3-isobutylxanthine. Cytotoxicity of alpha, beta-Amyrin was assessed by MTT assay. Lipid content in adipocytes was determined by Oil-Red O staining. In addition, the protein expression levels of peroxisome proliferator-activated receptor gamma (PPARgamma), CCAAT/enhancer binding proteins alpha (C/EBPalpha), beta (C/EBPbeta), and delta (C/EBPdelta) and glucose transporter 4 (GLUT4) were determined by qRT-PCR and western blot analysis. Oil-Red O staining revealed markedly reduced fat accumulation by alpha, beta-Amyrin (6.25--50 mug/mL) without affecting cell viability. Furthermore, our results indicate that alpha, beta-Amyrin can significantly suppress the adipocyte differentiation by downregulating the expression levels of adipogenesis-related key transcription factors such as PPARgamma and C/EBPalpha, but not C/EBPbeta or C/EPBdelta. In addition, the protein expression of membrane GLUT4 in 3T3- L1 adipocytes treated with alpha, beta-Amyrin was significantly higher than in control cells, indicating that alpha, beta-Amyrin augments glucose uptake. These findings suggest that alpha, beta-Amyrin exerts an anti-adipogenic effect principally via modulation of lipid and carbohydrate metabolism in 3T3-L1cells. The present in vitro findings, taken together with our earlier observation of the anti-obesity effect in vivo, suggest that alpha, beta-Amyrin can be developed as a new therapeutic agent for treatment and prevention of obesity. Author Affiliation: (a) Postgraduate Program in Pharmacology, Faculty of Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil (b) Postgraduate Program in Medical Sciences, Faculty of Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil (c) Research Center on Biodiversity and Biotechnology, Federal University of Piaui, Parnaiba, Piaui, Brazil (d) Department of Organic Chemistry, Federal University of Piaui, Teresina, Piaui, Brazil (e) Department of Physiology and Pharmacology, Natural Products Laboratory, Faculty of Medicine, Federal University of Ceara, Fortaleza, Ceara, Brazil * Corresponding author at: Department of Physiology and Pharmacology, Natural Products Laboratory, Faculty of Medicine, Federal University of Ceara, Cel Nunes de Melo, 1315, Fortaleza, Ceara, 60430-270, Brazil. Article History: Received 19 June 2018; Revised 3 November 2018; Accepted 6 November 2018 Byline: Karina Moura de Melo (a), Francisca Tuelly Bandeira de Oliveira (b), Rose Anny Costa Silva (b), Ana Luiza Gomes Quindere (b), Jose Delano Barreto Marinho Filho (c), Ana Jersia Araujo (c), Eanes Delgado Barros Pereira (b), Adonias Almeida Carvalho (d), Mariana Helena Chaves (d), Vietla Satyanarayana Rao (e), Flavia Almeida Santos fasufc@gmail.com (e,*)