This study aims to explore the relationship between Sirt3 expression and lipid accumulation in macrophages by inducing mitochondrial IDH2 deacetylation. In this study, Sirt3 interference and overexpression lentiviral vectors were constructed. Macrophages collected from C57BL/6J mice by peritoneal lavage were used to construct Sirt3 gene interference and overexpression models, and cultured in medium containing 1 mg/ml ox-LDL for 72 h to observe the enrichment of ox-LDL. Reverse transcription PCR was used to detect the expression of Sirt3 mRNA, western blot to detect Sirt3 and acetylated IDH2 proteins, and Nile Red staining and flow cytometry to detect intracellular lipids in macrophages. The results indicated that as compared to Sirt3 overexpressed and normal groups, the acetylation of IDH2 and accumulation of ox-LDL were significantly higher in the Sirt3 inhibited group. In conclusion, the expression of Sirt3 can inhibit lipid accumulation in macrophages by inducing mitochondrial IDH2 deacetylation.