About 40% of dairy cattle develop uterine disease at postpartum, causing infertility. Some studies indicate that uterine infection, predominantly by Escherichia coli in the first week postpartum, is associated with metritis. Metritis is an inflammation of the uterus in which the cow fails to completely clear bacterial contaminants and reducing postpartum inflammatory processes. The aim of this study was to evaluate the potential role of 4 lactic acid bacteria (LAB; Lactobacillus rhamnosus, Pediococcus acidilactici, Lactobacillus reuteri, and Lactobacillus sakei) in the modulation of Escherichia coli infection and its respective inflammation response by endometrial cells. Primary endometrial epithelial cells were isolated from a fresh endometrium of a healthy cow and cultured at 8 × 10⁴ cells/well in 24-well plates to evaluate the effects of LAB at 3 different doses. Cell extracts were obtained with TriZol. Pro-inflammatory status was assessed by qPCR quantification of IL8, IL1β, TNFα, and IL6 gene expression. Internalization of Escherichia coli was determined by direct enumeration on MacConkey agar plates. Data were normalized and analyzed by an ANOVA. Lactobacillus sakei and Lactobacillus reuteri had a positive effect preventing E. coli infection (87% and 78% respectively, P < 0.001) but were associated with a dose-variable effect on tissular inflammation that could further exacerbate the pro-inflammatory status of the endometrium. Pediococcus acidilactici clearly decreased (P < 0.001) E. coli internalization up to an 83% reduction and slightly reduced the inflammation (up to 3.2-fold in IL1β expression, P < 0.001). However, the pro-inflammatory cytokines IL8 and IL1β decreased significantly (P < 0.001) up to 85.1 and 5.2 folds, respectively, in the presence of L. rhamnosus. In conclusion, these results demonstrate a clear beneficial effect of P. acidilactici and L. rhamnosus in the modulation of endometrial infection and inflammation in cattle.