Low Oscillating Shear Stress (LOSS) is known to promote inflammation and atherosclerosis, impacting the phenotype and function of Vascular Endothelial Cells (VECs). The microphthalmia/transcription factor E (MiT/TFE) family of transcription factors regulates autophagy and lysosomal biogenesis in response to internal and external stressors. However, whether LOSS affects the expression of MiT/TFE family members in Human Umbilical Vein Endothelial Cells (HUVECs) has remained unclear. In this study, we utilized qPCR and WB to examine how LOSS influenced the mRNA and protein expression of MiT/TFE family members (MITF, TFEB, TFEC, and TFE3) in HUVECs in vitro. The in vitro findings were further confirmed using immunohistochemistry in a mouse model with a partially ligated carotid artery. Our results showed that LOSS down-regulated the expressions of TFEC and TFE3, while having no impact on MITF and TFEB expressions in HUVECs, the down-regulated expression of TFE3 were validated in the animal model. In conclusion, this study systematically explored the expression patterns of the MiT/TFE family in VECs under LOSS, providing new insights into the molecular mechanisms of abnormal hemodynamics-induced vascular endothelial inflammation-related diseases like atherosclerosis.