Procyanidins have lipolysis effect on adipose metabolism, but the underlying mechanism is not fully understood. The aim of present study was to examine the effect of grape seed procyanidin B2 (GSP) on the adipogenic differentiation of 3T3-L1 preadipocyte cell line and investigate the underlying mechanism. The results showed that GSP treatment significantly reduced the intracellular lipid accumulation in induced 3T3-L1 cells by targeting miR-483-5p as well as peroxisome proliferator-activated receptor γ (PPARγ). In addition, our results revealed that overexpression of miR-483-5p increased adipogenic differentiation, while inhibition of miR-483-5p reduced the lipid accumulation by suppressing the adipogenic differentiation. Moreover, overexpression of miR-483-5p could reverse GSP’s inhibition of adipocyte differentiation as well as increase the level of PPARγ. These results demonstrate that GSP inhibits adipogenesis by targeting PPARγ and suggest this effect might be mediated by miR-483-5p.