Microplastics (MPs), as a new environmental pollutant, have received widespread attention worldwide. Uterine fibrosis is one of the main factors of female reproductive disorders. However, it is unclear whether the female reproductive disorders caused by MPs are related to uterine fibrosis. Therefore, in this study, we constructed female mouse models exposed to polystyrene microplastics (PS-MPs). We found that PS-MP exposure resulted in endometrial thinning and severe collagen fibre deposition in female mice. Further mechanistic studies found that PS-MP exposure increased the expression of high mobility group Box 1 (HMGB1) and acetyl-HMGB1, further activating the Toll-like receptor 4/NADPH oxidase 2 (TLR4/NOX2) signalling axis and eventually causing oxidative stress . Afterwards, oxidative stress elicited the activation of Notch and the transforming growth factor β (TGF-β) signalling pathway , leading to increased levels of fibrotic proteins and collagen. Correspondingly, PS-MP treatment upregulated the expression of TLR4 and NOX2 and the level of reactive oxygen species (ROS) and increased the levels of fibrotic protein and collagen in mouse endometrial epithelial cells cultured in vitro. Conversely, inhibition of the TLR4/NOX2 signalling pathway effectively reduced the level of ROS in cells, weakened the upregulation of Notch and TGF-β signalling by PS-MPs, and efficiently reduced the expression of fibrotic and collagen genes. In summary, we demonstrated a new mechanism by which MPs induce uterine fibrosis in mice, that is, by inducing oxidative stress to activate the Notch and TGF-β signalling pathways by triggering the TLR4/NOX2 signalling axis. Targeting TLR4/NOX2 signalling may consequently prove to be an innovative therapeutic option that is effective in alleviating the reproductive toxicity of PS-MPs. Our study sheds new light on the reproductive toxicity of MPs and provides suggestions and references for comparative medicine and clinical medicine.