Objective To study the role of abnormally changed migration of vascular smooth muscle cells (VSMCs) induced by low shear stress (LowSS) in vascular remodeling during atherosclerosis as well as the molecular mechanism involved in this process. Methods By using comparative proteomic analysis with two-dimensional electrophoresis combined with mass spectrometry, differential protein expression profiles of cultured vascular tissues under normal shear stress (NSS) (1.5 Pa) and LowSS (0.5 Pa) were studied. Using endothelial cells (ECs) and VSMCs co-cultured parallel plate flow chamber system, two levels of shear stress i.e. LowSS and NSS, were applied, respectively. Western blot was used to detect the protein expressions of Rab28 and phosphor-ERK. Transwell system was used to detect the migration ability of VSMCs. After using RNA interference and ERK inhibitor PD98059 to decrease the expressions of Rab28 and phosphor-ERK, respectively, the migration ability of VSMCs was observed again. Results The expression of Rab28 in the cultured rat aorta was significantly up-regulated by the LowSS (0.5 Pa) application in comparison with the NSS (1.5 Pa). The migration, expressions of Rab28, and phosphorylation of ERK in VSMCs were significantly increased by the LowSS application. Target RNA interference of Rab28 significantly decreased the migration of VSMCs, but had no specific effect on the phosphorylation of ERK. Target inhibitor of ERK, PD98059, significantly decreased both the migration and Rab28 expression in VSMCs. Conclusions The LowSS may increase the phosphorylation of ERK and then increase the expression of Rab28 in VSMCs, which subsequently modulate VSMC migration during vascular remodeling. The investigation on the role of Rab28 and its signal path in LowSS-regulated VSMCs as well as the molecular mechanism might provide a biomechanical reference for understanding the pathogenesis of vascular remodeling during atherosclerosis and finding the therapeutic target of new drugs.