Background: GPR41 (G protein-coupled receptor 41) is a mammalian GPCR, which is activated by short-chain fatty acids (SCFAs) such as acetate (C2), propionate (C3) and butyrate (C4). GPR41 activation is known to involve numerous actions including enhancement of intestinal hormone secretion, leptin secretion and sympathetic activity. Objective: This study aimed to prove the ability of GPR41 activation to decrease lipid accumulation using selective GPR41 agonists, AR420626 and 1-methylcyclopropane-1-carboxylic acid (1-MCPC), in 3T3-L1 adipocytes. Materials and Methods: The 3T3-L1 adipocytes were differentiated from 3T3-L1 preadipocytes. Cell viability was performed by MTT assay and lipid accumulation was analyzed by Oil red O staining assay. Peroxisome proliferator-activated receptor gamma (PPAR-γ) expression was detected by Western blotting. Results: AR420626 and 1-MCPC significantly reduced lipid accumulation and decreased PPAR-γ expression in a dose-dependent manner. These effects were abolished by both knock-down of GPR41 treated with siRNA and pertussis toxin, an inhibitor of Ga_i-mediated signalling. Conclusion: These findings indicate that these compounds used in this study exert the blockade of lipid accumulation in 3T3-L1 adipocytes through Ga_i-coupled GPR41 pathways, resulting in the reduction of PPAR-γ expression.