Mastitis is one of the most critical diseases of dairy herds, with around 15–35% of clinical cases that still have unknown etiology. We hypothesized that the transfer of endogenous bacterial component is one of the possible causes of mastitis. This study aimed to determine whether intrauterine-infused lipopolysaccharide (LPS) can be translocated into the mammary gland and induces the inflammatory responses. Eighteen goats were used in this experiment. Sixteen goats were divided into 2 groups: infused with 5 mL of saline with or without 100 µg of E. coli LPS into uterus, and 2 other goats were LPS-infused and used for mammary gland tissues collection. Milk and blood were collected at 0 - 144 h after infusion to measure the blood leukocyte count (BLC), plasma LPS-binding protein (LBP), milk yield, milk somatic cell count (SCC), and pro- and anti-inflammatory cytokines (IL-1β, TNFα, IL-8, IL-10 in plasma and IL-1β, TNFα, IL-6, IL-8 in milk). The enzyme immunoassay was conducted to measure the cytokines and LBP. Mammary gland tissues were collected in the parenchyma area before and at 24 h after LPS infusion by biopsy, and the localization of LPS was observed by immunohistochemistry. Statistical analysis was completed using SAS software (version 9.4). Data were assessed for normality using the UNIVARIATE procedure and the basal values were analyzed by ANOVA. The repeated-measures analysis was used to analyze continuous data using the MIXED procedure. This model included treatment, time, the interactions between treatment and time, and the basal values as a covariate. Statistical significance of the contrast analysis was defined as P < 0.05. The mean of BLC (P = 0.053), plasma LBP (P = 0.058) and TNFα (P = 0.061) tended to be affected by the interaction between treatment and time, while in milk, the mean concentration of IL-1β (P = 0.004) and IL-6 (P = 0.017) were significantly elevated compared with the control group. The LPS was not detected in the deep area of the mammary gland before infusion, but in the connective tissues and interepithelial spaces of alveoli of the mammary gland at 24 h after LPS infusion. We conclude that intrauterine-infused LPS can be translocated to the mammary gland and induced inflammatory responses.