Objective To investigate the effects of umbilical cord-derived mesenchymal stem cells on the expression of sex hormones CYR61 and CTGF in rabbits with premature ovarian failure, and to reveal the treatment effects and associated mechanism. Methods Ten rabbits were randomly divided into a model group and a treatment group. The model of premature ovarian failure ( POF) was established by intraperitoneal injection of CTX [ 50 mg / ( kg·d), 2 consecutive days]. The treatment group was treated with UCMSCs (5×106 / ml, 2 ml each time for 3 days), and the model group was injected with the same amount of sterile water. Venous blood was taken at 0, 7, 14, and 28 days after UCMSC treatment, and analyzed for changes in hormone levels by ELISA. Ovarian tissue samples were taken 28 days after UCMSC treatment to detect CYR61 and CTGF proteins and their associated mRNA expression. Results There were no differences in E2 and FSH levels between the two groups before treatment (P>0. 05). After treatment, E2 level in the treatment group increased and the level of FSH decreased. Compared with the model group, the difference was statistically significant (P< 0. 05), and in the treatment group, the level of sexual hormone fluctuated periodically with time. Meanwhile, the model group rabbits always had a low E2 level and a high FSH level, and they did not show periodic fluctuations over time. Also, compared with the model group, the expression of CYR61 and CTGF in the treatment group was significantly increased, and the difference was statistically significant (t_CYR61 = 8. 688, P<0. 05; t_CTGF = 10. 142, P<0. 05). In addition, compared with the model group, the expression levels of CYR61 and CTGF mRNA in the treatment group were significantly increased, and the differences were statistically significant (t_CYR61= 38. 156, P<0. 05; t_CTGF = 14. 646, P<0. 05). CYR61 and CTGF were mainly expressed in the granulosa cells and follicular cell membranes. Compared with the model group, this expression of the treatment group was significantly increased, and the difference was statistically significant ( t_CYR61 = 3. 981, P<0. 05; t_CTGF = 4. 111, P<0. 05). Conclusions Umbilical cord-mesenchymal stem cell therapy can improve sex hormone levels, and upregulate the expression of CYR61 and CTGF in rabbits with POF, thereby activating sleep follicles, restoring ovarian function, and treating POF.