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This study investigated the effect of Luoshi Neiyi Formula(LSNYF) on hypoxia-inducible factor-1α(HIF-1α) and steroidogenic factor 1(SF-1) in endometriosis(EMs), aiming to explore the mechanism of Luoshi Neiyi Formula in treating EMs. Immunohistochemistry and quantitative real-time PCR(qPCR) were employed to determine the expression of HIF-1α and SF-1 in the endometriotic tissue. Human primary endometrial stromal cells(ESCs) were extracted and identified, in which the expression levels of HIF-1α and SF-1 were measured by immunofluorescence and qPCR. Cobalt chloride(CoCl_2) was used to simulate a low-oxygen environment in vitro, and the lentivirus with down-regulated expression of HIF-1α was constructed. The expression of HIF-1α or SF-1 was then determined by qPCR or Western blot. The effects of Luoshi Neiyi Formula-containing serum on the proliferation, migration, estradiol(E_2), HIF-1α, and SF-1 of ESCs in the low-oxygen environment were observed. Finally, the rat model of EMs with the syndrome of Qi stagnation and blood stasis was established by multi-factor intervention combined with autologous endometrium transplantation, and the effect of Luoshi Neiyi Formula on HIF-1α and SF-1 in the endometriotic tissue were studied. Compared with the normal endo-metrial tissue and ESCs, the endometriotic tissue and ectopic ESCs presented up-regulated mRNA and protein levels of HIF-1α and SF-1. Compared with the control group, treating eutopic ESCs with 50 μmol·L~(-1) CoCl_2 for 24 h promoted cell proliferation and up-regulated the mRNA level of HIF-1α and protein level of SF-1, while knocking down HIF-1α down-regulated the expression of SF-1 and E_2. Compared with the model group, the Luoshi Neiyi Formula-containing serum inhibited the proliferation and migration of ESCs and down-regulated the expression of E_2, HIF-1α, and SF-1. Similarly, the treatment with Luoshi Neiyi Formula attenuated the pathological injury of the endometriotic tissue and down-regulated the mRNA and protein levels of HIF-1α and SF-1 in the rat model of EMs. In summary, LSNYF may treat EMs by alleviating hypoxia and inhibiting the HIF-1α/SF-1 axis.