Melatonin regulates autophagy in granulosa cells from patients with premature ovarian insufficiency via activating Foxo3a.

Premature ovarian insufficiency (POI) is a diverse form of female infertility characterized by a decline in ovarian function before the age of 40. Melatonin (MT) is a potential clinical treatment for restoring or safeguarding ovarian function in POI. However, the specific therapeutic mechanism underlying this effect remains unclear. To address this, we conducted experiments using human granulosa cells (GCs) from both POI and normal patients. We examined the expression levels of autophagy-related genes and proteins in GCs through qRT-PCR and western blot analysis. Autophagy flux was monitored in GCs infected with GFP-LC3-adenovirus, and the regulatory function of MT in autophagy was investigated. Additionally, we employed pharmacological intervention of autophagy using 3-Methyladenine (3-MA) and RNA interference of Forkhead box O-3A (FOXO3A) to elucidate the mechanism of MT in the autophagy process. Compared to GCs from normal patients, GCs from POI patients exhibited irregular morphology, decreased proliferation, increased apoptosis, and elevated ROS levels. The expression of autophagy-related genes was downregulated in POI GCs, resulting in reduced autophagic activity. Furthermore, MT levels were decreased in POI GCs, but exogenous MT effectively activated autophagy. Mechanistically, melatonin treatment downregulated FOXO3A expression and induced phosphorylation in POI GCs. Importantly, silencing FOXO3A abolished the protective effect of melatonin on GCs. These findings indicate that autophagy is downregulated in POI GCs, accompanied by a deficiency in MT. Moreover, we demonstrated that supplementing MT can rescue autophagy levels and enhance GC viability through the activation of FOXO3A signaling. Thus, MT-FOXO3A may serve as a potential therapeutic target for POI treatment.

[Acupuncture-moxibustion treatment by stages based on the theory of "transformation of yin and yang" for premature ovarian insufficiency].

OBJECTIVE: To compare the efficacy between acupuncture-moxibustion treatment by stages and femoston for premature ovarian insufficiency (POI). METHODS: A total of 66 patients with POI were randomly divided into an observation group (33 cases, 3 cases dropped off) and a control group (33 cases, 2 cases dropped off). The patients in the observation group, based on the theory of "transformation of yin and yang", were treated with acupuncture-moxibustion by stages in the postmenstrual period, ovulatory period, premenstrual period and menstrual period, once every other day, 3 times a week. The patients in the control group were treated with oral administration of femoston (estradiol tablets/estradiol and dydrogesterone tablets, 1 tablet per day). Both groups were treated for 3 menstrual cycles. The ovarian function (serum follicle-stimulating hormone [FSH], luteinizing hormone [LH], estradiol [E2], anti-mullerian hormone [AMH] and antral follicle count [AFC]) and perimenopausal symptoms [modified Kupperman index (KI) scale score] were observed before and after treatment, and the menstrual improvement of the two groups was compared. RESULTS: Compared before treatment, the serum levels of FSH and LH were decreased (P<0.01), the levels of E2 were increased (P<0.01) in the two groups after treatment; the serum level of AMH and AFC in the observation group were increased after treatment (P<0.01). After treatment, the serum level of AMH and AFC in the observation group were higher than those in the control group (P<0.05). After treatment, there was no significant difference in the menstrual return rate and menstrual regularity rate between the amenorrhea patients of the two groups (P>0.05). After treatment, the modified KI scale scores in the two groups were reduced (P<0.01), and the modified KI scale score in the observation group was lower than that in the control group (P<0.05). CONCLUSION: Acupuncture- moxibustion treatment by stages based on the theory of "transformation of yin and yang" has similar efficacy with femoston in improving serum sex hormone level and menstrual symptoms in patients with POI, and has advantages in improving serum AMH level, AFC and perimenopausal symptoms.