Objective: To explore the dose-time dependent relationship of shear stress and different exposure times on the expression of thrombospondin-4 (THBS4) in human aortic smooth muscle cells (HASMCs), and to reveal its potential mechanism in vascular remodeling, with the aim of guiding the treatment of clinical cardiovascular diseases. Methods: HASMCs were subjected to shear stress of 0 dyn/cm2、4 dyn/cm2, 8 dyn/cm2, and 12 dyn/cm2 using a parallel plate flow chamber for 6 hours and 12 hours, respectively. Cells were collected after the treatment. The mRNA expression of THBS4 was detected by real-time fluorescence quantitative PCR, and the protein expression was detected by Western Blot. Immunofluorescence staining was performed to determine the co-localization of THBS4 and cleaved- Caspase 8. Results: The mRNA and protein expression of THBS4 gradually increased with the increase of shear stress and the extension of time. The expression levels of the gene and protein in the 12 dyn/cm2 group were significantly higher than those in the 0 dyn/cm2, 4 dyn/cm2, and 8 dyn/cm2 groups. The immunofluorescence results demonstrated that the expression of THBS4 was co-localized with cleaved- Caspase 8 and exhibited consistent expression levels. Conclusion: The spatiotemporal gradient changes of shear stress can regulate the expression of THBS4 in HASMCs, and its expression level is positively correlated with the magnitude of shear stress and has a time-dependent cumulative effect. THBS4 may modulate apoptotic pathways in HASMCs under shear stress.