Abstract: Objective To comprehensively investigate the potential impact of interleukin-10 (IL-10) overexpression on macrophage polarization and their targeting capabilities towards atherosclerotic plaques through transcriptome sequencing and ex vivo tissue imaging techniques. Methods Transcriptome sequencing and differential gene analysis were performed after infecting macrophages with a lentivirus to induce overexpression of IL-10 (IL-10M). The changes in expression levels of macrophage polarization markers were validated using RT-qPCR and flow cytometry. Subsequently, ApoE mice were intravenously administered IL-10M and ConM via the tail vein after being fed a Western diet, and the targeting efficiency of cells towards aortic plaques was assessed through ex vivo and histological fluorescence imaging. Results Transcriptomic analysis revealed that there were 1271 differentially expressed genes in IL-10M compared to the control group (ConM). The IL- 10M group exhibited significant expression changes in several key genes associated with macrophage polarization. M1 phenotype markers Cd86 and tumor necrosis factor (Tnf) exhibited downregulation, while Il-1beta demonstrated upregulation in IL-10M; M2 phenotype marker mannose receptor 1 (Mrc1) displayed upregulation, whereas arginase-2 (Arg2) showed downregulation. RT- qPCR confirmed the expression of Mrc1, Tnf- alpha, and Il-1beta in IL-10M, consistent with the sequencing results. Furthermore, flow cytometry revealed a reduction in CD86 expression levels in IL-10M compared to ConM. After intravenous injection, fluorescence signals of IL-10M and ConM can be respectively observed at the site of atherosclerotic plaque in mice aorta. Conclusion The overexpression of IL-10 modulated the expression of markers associated with macrophage polarization while preserving their targeting ability towards plaques. This discovery establishes a scientific foundation for further investigation into the potential role of IL-10 in preventing and treating atherosclerosis, thereby supporting the development of novel anti-inflammatory therapeutic strategies.