Background: Current in vitro vascular models are too simple compared with the real vascular environment. In this research, a novel in vitro 3D vascular disease model that simulated the vascular microenvironment was introduced. Methods: This model was mainly established by low shear stress and co-culture of endothelial cells and smooth muscle cells. Characterization and reproduction of the pathological state of the 3D model were determined. The effect of two clinical drugs was verified in this model. The difference of drug screening between a traditional oxidative-damaged cell model and this 3D model was determined by HPLC. Results: This model presented many disease markers of vascular diseases: abnormal cellular shape, higher endothelial cell apoptotic rate and smooth muscle cell migration rate, decreased superoxide dismutase level, and increased malondialdehyde and platelet-derived growth factor level. The drugs effectively reduced the disease indices and relieved the damage caused by low shear stress. Compared to the traditional oxidative-damaged cell model, this 3D model screened different active components of Salviae Miltiorrhizae extract, and it is closer to clinical studies. Conclusions: These results suggest that the 3D vascular disease model is a more efficient and selective in vitro study and drug screening platform for vascular diseases than previously reported in vitro vascular disease models.