Aim To explore the mechanism of adipophilin regulating neutral cholesterol ester hydrolase (nCEH) expression thus mediating lipid accumulation in RAW264.7 macrophages. Methods The constructed retroviral plasmid vectors with adipophilin of no expression and over expression were transfected into package cell PA317, then collected virus solution was used to infect RAW264.7 macrophages by screening and identifying to obtain RAW264.7 cell lines with adipophilin stably silenced and highly expressed. The mRNA and protein expression levels of nCEH were measured by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot respectively. High performance liquid chromatography (HPLC) was taken to detect the contents of intracellular cholesterol ester. Results The cholesterol and cholesterol ester levels were significantly higher in adipophilin over-expression group than in the control group (P<0.01), while dramatically lower than that in the control group when adipophilin was supressed (P<0.01). In addition, adipophilin inhibited nCEH expression. Furthermore, in RAW264.7 macrophages with high adipophilin expression, treatment with 100 nmol/L protein kinase Cδ agonist PMA for 30 min resulted in remarkable reductions of nCEH mRNA and protein expression(P<0.05), on the contrary, treatment with 100 nmol/L PKCδ inhibitor Rottlerin for 30 min increased the expression of nCEH(P<0.05). Conclusion Adipophilin may exert influence over lipid accumulation by inhibiting the expression of nCEH in RAW264.7 macrophages, and PKCδ played an important role in this regulatory mechanism.