Endometritis is a common disease in postpartum cows characterized by delayed uterine recovery due to inflammation of the endometrium. Although antibiotics and hormones are commonly used for treatment, they have their limitations. One potential alternative is the use of motherwort extract, specifically leonurine, which has been found to possess anti-inflammatory properties. However, the exact molecular mechanism of action remains unclear. To address this gap in knowledge, this study aimed to investigate the effects of leonurine on LPS-induced endometritis in mice using an experimental mouse model. Through transcriptomic analysis, it was discovered that leonurine modulates multiple signaling pathways, including JAK-STAT/PI3K-Akt, and influences the expression of key genes such as PRLR, SOCS2, COL1A1, and AKT1. Furthermore, leonurine effectively reduces the levels of inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1beta (IL-1β), which play a crucial role in the regulation of acute endometritis. Additionally, leonurine helps maintain cholesterol homeostasis and attenuates inflammation through the PPAR signaling pathway by modulating genes like CYP27A1, HMGCS1, and SCD2. These findings suggest that leonurine has a protective effect against LPS-induced endometritis and its anti-inflammatory properties involve multiple pathways and targets, potentially mediated by the regulation of signaling pathways such as JAK-STAT/PI3K-Akt/PPAR.