[Abstract] Objective: To explore the effect of Dingkun Pill on PI3K/Akt/mTOR signaling pathway in rats with endometriosis (EMs). Methods: The EMs rat model was constructed by heterotopic transplantation of endometrial tissue and randomly divided into five groups: model group (M group), Dingkun Pill low (1.13g/kg) dose group (DKP-L group), Dingkun Pill medium (2.26g/kg) dose group (DKP-M group), Dingkun Pill high (4.52g/ kg) dose group (DKP-H group), gestrinone (60mg/kg) group (GES group), with 12 rats in each group. Another 12 normal SD rats were opened the abdomen without transplantation ectopic endometrial tissue, and set as sham operation group (sham group). The rats were killed after intervention with drugs, the volume of ectopic endometrium and the weight of ectopic lesions of rats in each group were measured, the microvessel density (CD31 positive rate) and the expression of VEGF and MMP-9 in rat ectopic endometrial tissue were detected by immunohistochemical staining; the rat serum VEGF, MMP-9, iNOS and TNF-α levels were detected by enzyme-linked immunosorbent assay (ELISA); the expression of PI3K/Akt/mTOR pathway related proteins in rat ectopic endometrial tissue was detected by western blotting. Results: Compared with the sham group, the microvessel density, VEGF and MMP-9 expression, serum VEGF, MMP-9, iNOS and TNF-α levels, p-PI3K/PI3K, p-Akt/Akt, and p-mTOR/mTOR in the ectopic endometrial tissue were significantly increased in the M group (P<0.05); compared with the M group, the ectopic endometrial volume, the weight of the ectopic lesion, the microvessel density of the ectopic endometrial tissue, VEGF and MMP-9 expression, the serum VEGF, MMP-9, iNOS and TNF- α levels, p-PI3K/PI3K, p-Akt/Akt, p-mTOR/mTOR in ectopic endometrial tissue were all decreased in the drug intervention group, and DKP-L, DKP-M and DKP-H groups were dose-dependent (P<0.05); compared with the DKP-H group and the GES group, there was no significant difference in the indicators of rats (P>0.05). Conclusion: Dingkun Dan can reduce inflammation and inhibit ectopic endometrial growth in EMs rats, which may be achieved by blocking PI3K/Akt/mTOR signal pathway.