To observe the effect of Huayu Sanjie enema(HSE) on nuclear factor kappa B(NF-κB) pathway，inflammatory factors，and oxidative stress factors in endometriosis(EMT) rats with pelvic pain，and to explore the mechanism of HSE against pelvic pain in EMT.Methods:Of the 40 female SD rats，6 were randomly selected as the sham operation group with the random number table method.Autologous endometrial transplantation，estrogen，and oxytocin injection were employed to induce EMT with pelvic pain.Then 30 model rats were selected and randomized into model group，positive control group，and low-dose，medium-dose，and high-dose HSE groups，with 6 rats in each group.After 4 weeks of enema administration，the writhing response and pathological changes were observed.The expression of serum interleukin-1β(IL-1β)，tumor necrosis factor-α(TNF-α)，malondialdehyde(MDA)，and superoxide dismutase(SOD) and the expression of NF-κB protein in ectopic lesion tissue were detected.Results:Compared with the model group，low-dose，medium-dose，and high-dose HSE decreased the writhing times(P<0.05) and serum levels of IL-1β，TNF-α，and MDA(P<0.05)，raised SOD content(P<0.05)，and lowered NF-κB protein expression in ectopic lesion(P<0.05).Conclusion:HSE may inhibit the activation of NF-κB pathway by suppressing oxidative stress，thereby reducing the release of inflammatory factors and alleviating pelvic pain in EMT.