Moxibustion alleviates decreased ovarian reserve in rats by restoring the PI3K/AKT signaling pathway / 中西医结合学报

OBJECTIVE@#Moxibustion, a common therapy in traditional Chinese medicine, has potential benefits for treating decreased ovarian reserve (DOR). The present study investigates the protective effect of moxibustion in a rat model of DOR and explores the possible mechanisms.@*METHODS@#Sixty-four female Sprague-Dawley rats were randomly divided into four groups: control, DOR, moxibustion (MOX), and hormone replacement therapy (HRT). The DOR rat model was established by intragastric administration of 50 mg/kg Tripterygium glycoside suspension (TGS), once daily for 14 days. MOX and HRT treatments were given from the day TGS administration was initiated. The ovarian reserve function was evaluated by monitoring the estrus cycle, morphological changes in ovaries, levels of serum estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and anti-Mullerian hormone (AMH), pregnancy rate and embryo numbers. Terminal-deoxynucleotidyl transferase-mediated nick-end-labeling staining was used to identify ovarian granulosa cell apoptosis, while the protein and mRNA expressions of Bax, B-cell lymphoma-2 (Bcl-2), phosphatidylinositol 3-kinase (PI3K) and protein kinase B (AKT) in ovarian tissues were examined by immunohistochemistry, Western blot and quantitative reverse transcription-polymerase chain reaction.@*RESULTS@#Compared with the DOR group, MOX improved the disordered estrous cycle, promoted follicular growth, reduced the number of atresia follicles, increased the concentrations of serum E2 and AMH, and decreased serum FSH and LH concentrations. More importantly, the pregnancy rate and embryo numbers in DOR rats were both upregulated in the MOX treatment group, compared to the untreated DOR model. Further, we found that the MOX group had reduced apoptosis of ovarian granulosa cells, increased Bcl-2 expression and reduced expression of Bax. Furthermore, the PI3K/AKT signaling pathway was triggered by the moxibustion treatment.@*CONCLUSION@#Moxibustion improved ovarian function and suppressed apoptosis of ovarian granulosa cells in a rat model of DOR induced by TGS, and the mechanism may involve the PI3K/AKT signaling pathway.

Effect of acupuncture on oxidative stress and apoptosis in ovarian hypofunction induced by Tripterygium Glycosides / 针刺研究

OBJECTIVE: To explore whether manual acupuncture stimulation of "Shenshu " (BL23), "Guanyuan" (CV4), "Zhongwan" (CV12) can improve the ovarian function by resisting oxidant stress and reducing apoptosis of granulosa cells in rats with ovarian hypofunction. METHODS: Forty female SD rats with normal estrous cycles were randomly divided into blank control, model, hormone therapy and acupuncture groups (n=10 rats in each group). The ovarian hypofunction model was established by gavage of Tripterygium Glycosides suspension (50 mg·kg-1·d-1) for 14 successive days. Rats of the hormone therapy group were treated by gavage of estrogen and progesterone, and those of the acupuncture group treated by manual acupuncture stimulation of bilateral BL23 or CV4 and CV12 alternatively by using uniform reinforcing-reducing method for 10 s every 5 min (3 times in 10 min). The treatment was performed once daily for 14 days. The blank group was given equal volume of normal saline daily. On the 9th day, the estrous cycle of each rat was observed by means of vaginal smear test. The ovarian index, serum estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH), malondialdehyde (MDA) and superoxide dismutase (SOD) contents were detected by ELISA. Histopathological changes of the ovary tissue were observed by H.E. staining. The expressions of apoptosis-related proteins Bax and Bcl-2 and their mRNAs of the ovaries were determined by immunohistochemical staining and quantitative real time-PCR separately. RESULTS: The menstrual disorder rate was 100% in the model group, and was significantly higher than those in the hormone therapy (30%) and acupuncture (40%) groups (P<0.01). Following modeling, the ovarian wet weight and index, E2 and SOD contents, Bcl-2 protein and mRNA expressions were significantly decreased (P<0.01, P<0.05), and serum FSH, LH and MDA levels, Bax protein and mRNA expressions were significantly increased (P<0.05, P<0.01) in the model group in comparison with the blank control group. H.E. staining showed a large amount of connective tissue in the ovary, with fewer mature follicles, increase of atresia follicles, significant reduction of luteal tissue, and appearance of scarring tissue in the model group. Compared with the model group, there were more mature follicles, fewer atresia follicles, lower abnormal granulosa cell morphology and lower estrous cycle disorder ratio in both hormone therapy and acupuncture groups. After the interventions, the decreased ovarian wet weight and index, serum E2 and SOD contents, expressions of Bcl-2 protein and mRNA were significantly increased (P<0.05, P<0.01), and the increased levels of FSH and LH, MDA, expressions of Bax protein and mRNA were significantly decreased in both hormone therapy and acupuncture groups (P<0.05, P<0.01). The effect of acupuncture was significantly superior to that of hormone in up-regulating SOD, Bax and Bcl-2 protein expressions (P<0.01). CONCLUSION: Acupuncture can improve the menstrual disorder in rats with ovarian hypofunction, which is closely related to its effects in improving antioxidant stress ability and regulating the expressions of apoptosis-related protein and mRNA of ovarian gra-nulosa cells.