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The aim of current study is to investigate the molecular mechanism that caloric restriction (CR) suppresses endothelial cells senescence. Human aortic endothelial cells (HAECs) were divided into 5 groups: control group, high caloric group (about 1.5 times caloric intake of control group), low caloric group (about 0.5 times caloric intake of control group), siRNA plus low caloric group (low caloric treatment pretreated with special siRNA targeting hepatocyte nuclear factor 3γ (HNF3γ)), and siRNA plus high caloric group (high caloric treatment pretreated with special siRNA targeting HNF3γ). The gene and protein expressions of HNF3γ and NADPH oxidase 4 (NOX4) were quantified by real-time quantitative PCR (RT-qPCR) and Western blotting, respectively. Intracellular reactive oxygen species (ROS) production was measured by flow cytometry. Endothelial cells senescence was assayed by senescence associated β-galactosidase staining. After verifying the binding of HNF3γ to NOX4 promoter region by chromatin immunoprecipitation assay (ChIP), NOX4 promoter activity was assayed by dual-luciferase reporter system. Compared with the control group, the mRNA and protein expression levels of HNF3γ,and the ratio of phosphorylated HNF3γ protein increased significantly (P