| Objective:To observe effects of electroacupuncture combined with bone marrow mesenchymal stem cells on the treatment of rats chemotherapy-induced premature ovarian failure and explore the mechanisms of electroacupuncture on stem cells treatment.Methods:1.BMSCs were isolated from rat tibia and fibula for culture,and identify the expression level of the surface markers and identify osteogenic and adipogenic differentiation capabilities of BMSCs.2.An animal model of premature ovarian failure induced by chemotherapy in rats prepared by intraperitoneal injection of cyclophosphamide solution.3.After the preparation of the animal model,take a vaginal smear of the rat for 5consecutive days,and observe the changes of the estrus cycle by methylene blue staining to determine whether the model is successfully prepared.4.The animals were divided into normal group,POF group,stem cell group,and combined treatment group.BMSCs suspension containing 2×10~6cells was injected into the tail vein of rats in stem cell group and combined treatment group.The combined treatment group received electroacupuncture for 7 consecutive days.5.Serum hormones,ovarian tissue sections and follicle counts,BMSCs homing and granulosa cell apoptosis were detected in each group of rats after transplantation.Results:1.The shape of BMSCs cells is long spindle.The positive expression rates of CD29,CD44,CD90 and CD45 were 95.24%,98.55%,99.73%,2.79%,respectively detected by flow cytometry,and can differentiate into adipocytes and osteocytes under specific conditions.2.After modeling with cyclophosphamide,the rats showed reduced activity,reduced food and water intake,and loss of hair luster.The methylene blue staining of the vaginal smear found that the estrus cycle was disordered or delayed,which proved that the model preparation was successful.3.After treatment,the serum estradiol content of the rats in the two treatment groups was significantly increased,and the serum follicle stimulating hormone content was significantly decreased.(P<0.05).4.The histopathological results of the ovary found that the number of oocytes in the growth stage after treatment increased,and the structure of granular cells was obviously dense and the amount increased.5.BMSCs homing level detection revealed that the combined treatment group had more BMSCs homing to ovary than stem cell group(P<0.01),the expression level of chemokine receptor CXCR4 increased(P<0.05).6.After cyclophosphamide modelling,the apoptosis of granulosa cells in ovarian tissue was significantly increased.After treatment,apoptotic cells number in two treatment groups were reduced significantly,while the apoptotic cells number was even less in combined treatment group(P<0.05).Compared with the stem cell group,the expression of Bcl-2 protein in the ovary of the rats in the combined treatment group was increased(P<0.05),and the expression of Bax protein was decreased(P<0.05).Conclusion:1.The BMSCs were cultured by the bone marrow adherence method successfully.2.The intraperitoneal injection of cyclophosphamide can successfully prepare the chemotherapy-caused premature ovarian failure rat model.3.Electroacupuncture combined with BMSCs can achieve better results in the treatment of chemotherapy-type premature ovarian failure. |
