Premature Ovarian Failure Group Research Articles

SIRT1 overexpression by melatonin and resveratrol combined treatment attenuates premature ovarian failure through activation of SIRT1/FOXO3a/BCL2 pathway

AimTo evaluate the synergistic effect of combined treatment with melatonin (MEL) and resveratrol (RES) in cisplatin (CIS)-induced premature ovarian failure (POF) model in rats and to elucidate the molecular mechanism of this therapeutic effect. Material & methodsFemale Sprague Dawley rats were divided into 7 experimental groups as follows; CONT (Control), CIS, MEL, RES, POF + MEL, POF + RES, and POF + MEL + RES. H&E staining was performed to evaluate follicular cell vacuolization/degeneration, vascular congestion/hemorrhage, and inflammation, by using an ordinal scale from 0 to 4 to grade the severity of observed changes (0 = normal, 1 = mild, 2 = moderate, 3 = severe, 4 = very severe). Zona pellucida integrity and connective tissue amount in the ovarian tissue were detected using PAS & Masson Trichrome staining. The immunofluorescence method was used to determine the immune localizations of pH2Ax, SIRT1, FOXO3a, and BCL2. The connective tissue amounts and immunoreactivity staining intensities were measured using ImageJ. The gene expression of SIRT1, FOXO3a, and BCL2 was determined using RT-PCR. Serum estrogen hormone levels were measured by ELISA. Statistically, Bonferroni correction was performed, and p < 0.002 were considered significant. ResultsA significant difference was observed in the POF group compared to the CONT group in all parameters except tertiary follicle count and hemorrhage. The decrease in the number of atretic follicles in the POF + MEL + RES group was found significant compared to both POF + MEL and POF + RES groups. The expression of pH2Ax, SIRT1, FOXO3a, and BCL2 at the protein level and SIRT1 and BCL2 at the mRNA level were significant in the POF + MEL + RES group compared to the POF group. Between the single and combination treatment groups, the difference in protein level was found in pH2Ax, SIRT1, FOXO3a, and BCL2 expression. The POF + MEL + RES group exhibited significantly higher SIRT1 mRNA expression compared to the groups receiving single treatments. ConclusionThe present study provides evidence that MEL and RES have synergistic effects in preventing the decrease in follicle reserve and increase in DNA break (pH2Ax) and follicle atresia in POF ovaries. This therapeutic effect is mediated by SIRT1 overexpression and activation of the SIRT1/FOXO3a/BCL2 pathway.

Protective and/or therapeutic effects of berberine in a model of premature ovarian failure induced by cyclophosphamide in rats.

We conducted a biochemical and histological evaluation of whether berberine has a protective and/or therapeutic effect in a cyclophosphamide-induced premature ovarian failure (POF) model. We divided 28 Wistar albino female rats into 4 groups [control group, POF group, cyclophosphamide (CP)+berberine (Bb) group, and POF+Bb group]. The POF model was established by intraperitoneal administration of 50 mg/kg CP on day 1 followed by 8 mg/kg/day CP dissolved in saline for the following 14 days. The CP+Bb group received Bb concurrently for two weeks with CP. The POF+Bb group received berberine for two weeks following the completion of CP administration. Left ovaries were used for histopathological evaluation and right ovaries were used for biochemical analysis [tumor necrosis factor-alpha (TNF-α), interleukin (IL)-1, IL-6 levels in tissue]. Ovarian damage scoring was significantly higher in the POF group than in the other groups (p<0.005). In the POF group, primordial and primary follicle counts were the lowest, while secondary and corpus luteum counts were the highest (p<0.005). There was no significant difference between the other groups. The POF group had significantly elevated levels of TNF-α, IL-1, and IL-6 in the biochemistry results (p<0.005). We demonstrated that berberine could be effective in the protection and treatment of POF by reducing proinflammatory cytokines. We believe that our study can make a considerable contribution to the literature in terms of POF protection and/or treatment.

Synergistic Effects of Capsaicin and Quercetin Improved Induced Premature Ovarian Failure in Rat.

Premature ovarian failure (POF) is a heterogeneous disorder. POF is defined as hypergonadotropic hypoestrogenism in women under 40 years. There is no effective treatment to cure POF patients. Antioxidants prevent ovarian damage by reducing the lipid peroxidation cascades affecting folliculogenesis, meiosis and ovulation. Hence; the aim of present study was to investigate the effects of Capsaicin (CAP) and Quercetin (QUR) on cyclophosphamide (CYC)-induced POF in rat model. In this experimental study, POF was induced by intraperitoneal injection of 200 mg/kg CYC on first day and then 8 mg/kg/day for the following 3 days. After 4 days of CYC administration, rats were randomly divided into five groups (n=6/group) as follows: POF, dimethyl sulfoxide (DMSO), CAP (0.5 mg/kg/day), QUR (100 mg/kg/day) and CAP+QUR. Biochemical, hormonal, gene expression, and histological evaluations were performed on blood serum and tissue samples after 14 days of treatment with the CAP and QUR. CAP, QUR and CAP+QUR groups showed signs of restored ovarian function in the form of a significant increase in serum total antioxidant capacity (TAC), estrogen, progesterone and anti-mullerian hormone (AMH) levels versus POF and DMSO groups and a significant improvement in histological parameters and follicle numbers in treatment groups compared to POF and DMSO groups. Polymerase chain reaction (PCR) analysis demonstrated that CAP and QUR upregulate the expression of BAX gene and decreased the expression of apoptosis inducing genes (BCL-2 and P53). CAP and QUR treatment of CYC-induced POF rats showed a positive effect on reducing ovarian damage by improving TAC levels, expression of apoptotic genes, levels of ovarian reserve markers, and histological parameters. Our results suggest that treatment with CAP or QUR may be a conservative treatment approach for CYC -induced POF.

Hydrogen-rich water treatment targets RT1-Db1 and RT1-Bb to alleviate premature ovarian failure in rats.

Premature ovarian failure (POF) is defined as the cessation of ovarian function before the age of 40 years, imposing a significant health burden on patients. However, effective etiological therapy for POF is scarce. Thus, we aimed to explore the protective role and targets of hydrogen-rich water (HRW) in POF. Based on cyclophosphamide (CTX)-induced POF rat models, the protective role of HRW treatment was mainly determined through serum 17-β-estradiol (E2), follicle-stimulating hormone (FSH), anti-mullerian hormone (AMH) levels, ovarian histomorphological analysis, and TUNEL assay. Tandem mass tag (TMT)-based quantitative proteomic analysis was then conducted on ovarian tissues, and the targets of HRW in POF were identified integrating differential expression analysis, functional enrichment analysis, and interaction analysis. In HRW treatment of POF rats, the serum AMH and E2 levels significantly increased, and FSH level significantly reduced, indicating the protective role of HRW. After TMT quantitative proteomic analysis, a total of 16 candidate differentially expressed proteins (DEPs) were identified after the cross analysis of DEPs from POF vs. control and POF+HRW vs. POF groups, which were found to be significantly enriched in 296 GO terms and 36 KEGG pathways. The crucial targets, RT1-Db1 and RT1-Bb, were finally identified based on both protein-protein interaction network and GeneMANIA network. The HRW treatment could significantly alleviate the ovarian injury of POF rats; RT1-Db1 and RT1-Bb are identified as two crucial targets of HRW treatment in POF rats.

SİSPLATİN İLE İNDUKLENEN PREMATÜR OVER YETMEZLİĞİ SIÇAN MODELİNDE DİOSMİN UYGULAMASININ ETKİLERİ

m&#x0D; We aimed to examine the potential beneficial effects of diosmin administration on cisplatin - induced premature ovarian failure (POF) in a rat model&#x0D; &#x0D; Material and Methods&#x0D; Twenty-eight rats were divided into four groups. Group A rats (n:7) were determined as the sham group. The remaining rats received an intraperitoneal injection of 1.5 mg/kg/day cisplatin for 10 days to create a POF model. Then, they were randomly divided into 3 subgroups.&#x0D; Group B was determined as POF group. Group C rats were given 100mg/kg/day diosmin for 10 days simultaneously while creating POF model. Group D rats were given 100mg/kg/day diosmin for 10 days after POF model was created. Twentieth day blood samples were taken and left ovaries were resected for examination.&#x0D; &#x0D; Results&#x0D; CIS-induced rats showed reduced levels of SOD, AMH and E2 compared to sham group rats (p0.05) between the sham group, group C and D. No significant (p&gt;0.05) difference in FSH value was observed between group C, D and sham groups.&#x0D; There was no significant (p&gt;0.05) difference in the number of secondary and antral follicles between group C and D compared to the sham group. Primordial follicle count was significantly higher in group C than group B (p

Classical music restored fertility status in rat model of premature ovarian failure

Background:The restorative effect of classical music was assessed on the cyclophosphamide-induced animal model of premature ovarian failure (POF).Methods:Mozart’s piano classical music (K.448) was used for up to 4 and 8 weeks. Rats were exposed to music 6 h every day using a stereo system with a volume of 65–70 dB. Sera and ovarian tissue samples were collected for the evaluation of FSH, LH, and E2 and histopathological examination. At the same time points, samples were taken from the hypothalamus and hippocampus to monitor the expression of Ntrk2, Crh, and Pomc using real-time PCR. Mating trial was performed to evaluate the fertility status of POF rats.Results:Histopathological examination revealed a significant increase (p < 0.05) in the numbers of morphologically normal follicles at all the developmental stages in POF rats after music therapy compared to the POF group (p < 0.05). Music therapy decreased FSH and LH levels to near-to-normal levels conidied with elevation of E2 (p < 0.05). Ntrk2, Crh, and Pomc expressions were down-regulated in POF rats. Music therapy increasaed the expression of Ntrk2 in the hypothalamus of POF rats (p < 0.05). In contrast, Crh and Pomc failed to reach the detection limit before intervention and four weeks after the intervention however, these genes were expressed eight weeks after music therapy. Fertility status was increased (p < 0.05) in terms of litter size in POF rats after being exposed to music compared to the non-treated POF control group (p < 0.05).Conclusion:Results showed that music can exert therapeutic effects on POF rats via the alteration of sex-related hormones.

Study of Serum Anti-Mullerian Hormone in the Diagnosis of Ovarian Reserve Function in Patients with Premature Ovarian Insufficiency.

Objective To explore the diagnostic value of serum anti-Mullerian hormone (AMH) for patients with premature ovarian insufficiency (POI) and premature ovarian failure (POF). Methods Totally, 125 women with menstrual disorder treated in the Obstetrics and Gynecology Department of Ningbo Women & Children Hospital between January 2020 and December 2021 were enrolled. Among them, based on the follicle-stimulating hormone (FSH) level, 54 patients (25 IU/L < FSH ≤ 40 IU/L) were assigned to the POI group, and 71 patients (FSH > 40 IU/L) were assigned to the POF group. In addition, 72 individuals who went physical examination in the hospital and showed normal menstrual cycle were selected as the control (CON) group. Serum AMH in each group was quantified via enzyme-linked immunosorbent assay (ELISA) and Beckman Coulter Access active immunoassay analyzer, and the levels of serum neutral hormones [luteinizing hormone (LH), FSH, as well as estradiol (E2)] in each group were detected through the electrochemiluminescence method. The difference between AMH level acquired by the latest automatic method and that acquired by the traditional manual ELISA was compared, and the correlation of serum AMH with sex hormones was analyzed. In addition, receiver-operating characteristic (ROC) curves were drawn for determining the diagnostic value of serum AHM for POI ad POF. Results Beckman Coulter Access quantified AMH more accurately and fastly (Beckman Coulter Access: 35 minutes; manual quantification: 3-4 hours) and was more sensitive than ELISA, with a requirement to less serum. The levels of serum AMH and E2 in the POF group were 0.04 ± 0.10 ng/mL and 35.16 ± 53.06 ng/mL, respectively, which were notably lower than those in the POI group ((0.69 ± 1.46) ng/mL and (3.96 ± 2.82) ng/mL) and CON group ((76.31 ± 97.84) ng/mL and (113.19 ± 114.84) ng/mL). The LH and FSH levels in the POF group were 37.86 ± 19.44 IU/L and 75.05 ± 35.31 IU/L, which were higher than those in POI group ((22.66 ± 26.15) IU/L and (11.30 ± 17.05) IU/L) and the CON group ((29.81 ± 4.45) IU/L and (6.78 ± 3.45) IU/L) (P < 0.05). The POI group showed a notably lower serum AMH level and notably higher LH and FSH levels than the CON group (P < 0.05), and the POI group was similar to the CON group in the E2 level (P > 0.05). Serum AMH showed a positive correlation with E2 (r = 0.291, P < 0.05) and a negative association with both FSH (r = −0.476, P < 0.05) and LH (r = −0.143, P < 0.1). The optimal cut-off value of serum AMH in predicting POI was 0.83 ng/mL, and the corresponding sensitivity and specificity were 95.8% and 85.2%. The optimal cut-off of serum AMH in predicting POF was 0.075 ng/mL, and the corresponding sensitivity and specificity were 81.7% and 94.4%. The area under ROC curve (AUC) of serum AMH + FSH in the diagnosis of POF was close to 1. Conclusion Beckman Coulter Access AMH test is the latest automatic electrochemiluminescence sandwich immunoassay, with higher sensitivity and reproducibility than traditional manual ELISA and with ability to produce repeatable results. With the decline of ovarian function, the serum AMH of POI patients decreases gradually, and the serum AMH of POF patients decreases obviously, so serum AMH level has great value in predicting POI and POF and can be used as a sensitive index for early diagnosis of the two. Serum AMH combined with FSH can lift the diagnostic efficiency of POF.

The level distribution of serum AMH in different clinical manifestations of women of childbearing age

To investigate the distribution and diagnostic value of serum anti-Müllerian hormone (AMH) in healthy women of childbearing age, women with polycystic ovary syndrome (PCOS), ovarian dysfunction (DOR) and premature ovarian failure (POF). This study retrospectively selected female patients of childbearing age who were treated in the affiliated Obstetrics and Gynecology Hospital of Fudan University from January to December 2019. According to different clinical manifestations, they were divided into 133 cases in PCOS group, 120 cases in DOR group and 134 cases in POF group. 125 healthy women in the same period were selected as the control group. The values of serum AMH, estradiol (E2), follicle stimulating hormone (FSH), luteinizing hormone (LH) and testosterone (T) were measured in the four groups. Single sample Kolmogorov-Smirnov test, one-way ANOVA, independent sample t-test, Kruskal-Wallis H test, Mann-Whitney U test, logistic regression analysis and ROC curve were used for comparative analysis. The serum AMH levels of PCOS group, DOR group, POF group and control group were 9.10 (6.67, 11.49) ng/ml, 0.11 (0.05, 0.29) ng/ml, 0.03 (0.02, 0.06) ng/ml and 2.99 (1.57, 4.98) ng/ml, respectively [M(Q1,Q3)], the differences were statistically significant (P<0.001). The basal endocrine levels including E2, FSH, LH and T also had significant differences between groups (P<0.001). The results of multiple comparisons showed that there were significant differences in AMH and LH between DOR, POF and PCOS groups and the control group. The T level of PCOS group was significantly higher than that of the control group, the E2, LH and T levels of DOR group were significantly higher than that of the control group, and the FSH level of POF group was significantly higher than that of the control group (P<0.05). The area under the curve (AUC) of AMH and AMH+LH in the diagnosis of PCOS were 0.905 and 0.922, the sensitivity was 82.7% and 85.0%, and the specificity was 88.0% and 88.8%. The AUC of DOR was 0.861 and 0.971, the sensitivity was 89.0% and 92.5%, and the specificity was 63.0% and 92.0%. The AUC of POF was 0.950 and 0.998, the sensitivity was 98.3% and 99.2%, and the specificity was 75.9% and 97.0%, respectively. The AUC of AMH and AMH+LH combined indexes in the differential diagnosis of DOR and POF were 0.768 and 0.937, the sensitivity was 70.3% and 95.5%, and the specificity was 73.9% and 80.8%.

Comparison of CD146 +/− mesenchymal stem cells in improving premature ovarian failure

BackgroundMesenchymal stem cells (MSCs) are a heterogeneous group of subpopulations with differentially expressed surface markers. CD146 + MSCs correlate with high therapeutic and secretory potency. However, their therapeutic efficacy and mechanisms in premature ovarian failure (POF) have not been explored.MethodsThe umbilical cord (UC)-derived CD146 +/− MSCs were sorted using magnetic beads. The proliferation of MSCs was assayed by dye670 staining and flow cytometry. A mouse POF model was established by injection of cyclophosphamide and busulfan, followed by treatment with CD146 +/− MSCs. The therapeutic effect of CD146 +/− MSCs was evaluated based on body weight, hormone levels, follicle count and reproductive ability. Differential gene expression was identified by mRNA sequencing and validated by RT-PCR. The lymphocyte percentage was detected by flow cytometry.ResultsCD146 +/− MSCs had similar morphology and surface marker expression. However, CD146 + MSCs exhibited a significantly stronger proliferation ability. Gene profiles revealed that CD146 + MSCs had a lower levels of immunoregulatory factor expression. CD146 + MSCs exhibited a stronger ability to inhibit T cell proliferation. CD146 +/− MSCs treatment markedly restored FSH and E2 hormone secretion level, reduced follicular atresia, and increased sinus follicle numbers in a mouse POF model. The recovery function of CD146 + MSCs in a reproductive assay was slightly improved than that of CD146 - MSCs. Ovary mRNA sequencing data indicated that UC-MSCs therapy improved ovarian endocrine locally, which was through PPAR and cholesterol metabolism pathways. The percentages of CD3, CD4, and CD8 lymphocytes were significantly reduced in the POF group compared to the control group. CD146 + MSCs treatment significantly reversed the changes in lymphocyte percentages. Meanwhile, CD146 - MSCs could not improve the decrease in CD4/8 ratio induced by chemotherapy.ConclusionUC-MSCs therapy improved premature ovarian failure significantly. CD146 +/− MSCs both had similar therapeutic effects in repairing reproductive ability. CD146 + MSCs had advantages in modulating immunology and cell proliferation characteristics.

Evaluation of Serum AMH, INHB Combined with Basic FSH on Ovarian Reserve Function after Laparoscopic Ovarian Endometriosis Cystectomy.

ObjectiveThe value of serum AMH, INHB, and bFSH levels in assessing postoperative ovarian reserve function was analyzed by measuring serum anti-Mullerian hormone (AMH), inhibin B (INHB), and basal follicle-stimulating hormone (bFSH) levels in patients after laparoscopic cystectomy for endometrioma.MethodsFrom June 2019 to December 2021, 124 patients underwent laparoscopic cystectomy for endometrioma in our hospital were selected, and the serum AMH, INHB, bFSH level, antral follicle count (AFC) of all patients before and after operation were detected and compared. According to the results of postoperative testing, all the patients were divided into normal group (n = 86), diminished ovarian reserve (DOR) group (n = 27), and premature ovarian failure (POF) group (n = 11). Pearson correlation model and subject operating characteristic curve (ROC) were used to analyze the correlation and diagnostic value of serum AMH, INHB and bFSH levels with postoperative ovarian reserve function, respectively.ResultsAfter operation, the levels of serum AMH, INHB and AFC in the DOR group and POF group decreased compared with those before the operation, and the serum bFSH levels increased (p < 0.05). After operation, the levels of serum AMH, INHB and AFC in DOR group and POF group were lower than those in normal group,and the serum bFSH levels were higher than the normal group; the levels of serum AMH, INHB and AFC in POF group were lower than those in DOR group, and the serum bFSH levels were higher than the DOR group (p < 0.05). Pearson analysis showed that serum AMH and INHB levels were negatively correlated with bFSH, and positively correlated with the number of AFC, the serum bFSH level was negatively correlated with the number of AFC (p < 0.05). The diagnostic values of serum AMH, bFSH, INHB and the combination of the three tests for postoperative abnormal ovarian reserve function were 0.866 (95% CI, 0.801–0.923), 0.810 (95% CI, 0.730–0.890), 0.774 (95% CI, 0.687–0.860) and 0.940 (95% CI, 0.900–0.981), respectively.ConclusionSerum AMH and INHB levels decreased and bFSH levels increased in patients after laparoscopic cystectomy for endometrioma, both of which were closely related to postoperative ovarian reserve function, and both could evaluate ovarian reserve function after ovarian cyst debulking, and the combined test could significantly improve the detection rate.

Translation regulatory long non-coding RNA 1 (TRERNA1) sponges microRNA-23a to suppress granulosa cell apoptosis in premature ovarian failure

ABSTRACT Translation regulatory long non-coding RNA 1 (TRERNA1) plays critical roles in cancer biology. We predicted the direct interaction of TRERNA1 with microRNA (miR)-23a, which promotes granulosa apoptosis. Granulosa apoptosis is involved in premature ovarian failure (POF). This study was therefore carried out to explore the involvement of TRERNA1 and miR-23a in POF. The expression of TRERNA1 and miR-23a in POF and control groups were detected by RT-qPCRs. The subcellular locations of TRERNA1 in granulosa cell line COV434 was detected by subcellular fractionation assay. The interaction between TRERNA1 and miR-23a was predicted using IntaRNA2.0. The direct interaction between COV434 and miR-23a was explored with RNA pull-down assay. In granulosa cells, the direct interaction between TRERNA1 and miR-23a was verified by overexpression assay. Cell apoptosis assay was performed to evaluate cell apoptosis. Both TRERNA1 and miR-23a were downregulated in POF. In addition, TRERNA1 was detected in both cytoplasm and nuclear samples of granulosa cells, and directly interacted with miR-23a. TRERNA1 did not affect the expression of miR-23a in granulosa cells, while TRERNA1 suppressed the role of miR-23a in enhancing cell apoptosis. In conclusion, TRERNA1 may sponge miR-23a to suppress granulosa cell apoptosis in POF.

Prematür Overyan Yetmezlikli Ratlarda Yaş Kupa Terapisinin Anksiyete ve Tiyol/Disülfit Homeostazı Üzerine Etkileri: Deneysel Bir Çalışma

Objective: The effect of wet cupping therapy on anxiety and thiol/disulfide homeostasis was evaluated in rats with premature ovarian failure (POF) induced by cyclophosphamide (CYC). Material and Methods: Twenty four Wistar Albino female rats were divided into control (n=6), POF (n=6), wet cupping (n=6), POF+wet cupping (n=6) groups. CYC at a dose of 200 mg/kg was injected intraperitoneally to POF and POF+wet cupping groups. One week later, wet cupping therapy was applied to wet cupping and POF+wet cupping groups for 5 minutes. The next day, Open Field Test and Elevated Plus Maze Test were applied to all experimental groups to evaluate anxiety. After sacrification, to assess pattern formation, endometrial thickness and ovarian reserve were measured with hematoxylin-eosin staining. Thiol/disulfide homeostasis and erythrocyte oxidized-reduced glutathione levels were measured in serum to measure oxidative stress. Results: Although there was no statistical significance, it was observed that anxiety decreased in POF+wet cupping compared to POF group. There was no change in thiol-disulfide values in POF+wet cupping compared to POF group (p>0.05). Conclusion: Wet cupping therapy did not reduce anxiety in the experimental POF model and did not shift the balance to positive in thiol/disulfide homeostasis in favor of thiols. However, the obtained graphics, numerical data and high standard deviation suggest that meaningful data could be obtained with more subjects.

Protective effect of small peptides from Periplaneta americana on cyclophosphamide-induced premature ovarian failure.

To investigate the protective effect of small peptides from Periplaneta americana (SPPA) on cyclophosphamide (CP)-induced premature ovarian failure (POF) in mice. Silent mating type information regulation 2 homolog 1 (SIRT1) /tumor-associated protein 53 (p53) signaling pathway plays an important role in delaying POF. Hematopoietic progenitor cell antigen (CD34) reflects ovarian aging from the side. However, whether SPPA inhibits POF in mice by influencing the SIRT1/p53 pathway and CD34 expression remains to be studied. Forty female Kun Ming (KM) mice were divided into four groups: a control group (normal saline, n=10), POF model group (160 mg/kg CP, n=10), SPPA low-dosage group (160 mg/kg CP + 100 mg/kg SPPA, n=10), and SPPA high-dosage group (160 mg/kg CP + 200 mg/kg SPPA, n=10). CP administration route is intraperitoneal injection, and SPPA administration route is intragastric. Eyeball enucleation blood samples and the ovaries of mice were collected by midline laparatomy and oopherectomy, and the malondialdehyde (MDA), nitric oxide (NO), superoxide dismutase (SOD), follicle-stimulating hormone (FSH), and anti-Müllerian hormone (AMH) concentrations were tested. Immunohistochemical tests for the expressions of SIRT1, p53, and CD34 were carried out. Finally, ovarian mRNA levels of SIRT1 and p53 were detected with real-time fluorescence quantification PCR (qRT-PCR). A mouse model of POF was generated using 160 mg/kg of CP. Compared with POF group, we found that plasma NO, MDA, and FSH decreased, while AMH and SOD increased in the SPPA low-dose group. Compared with the POF group, the SPPA low- and high-dosage groups achieved significant growth in the number of primordial, primary, and total number of healthy follicles at all levels, but sharp reductions in the number of atretic follicles. In addition, we found downregulated protein and mRNA expression of SIRT1, and upregulated that of p53 were observed in ovarian tissues of treated mice with POF, in immunohistochemistry experiments and qPCR experiments. In contrast, high protein and mRNA expression of SIRT1, and low that of p53 were observed in SPPA treatment groups. And the results of CD34 protein expression were consistent with that of SIRT1. In total, SPPA significantly inhibited POF caused by CP in mice via activation of the SIRT1/p53 signaling pathway in the mouse ovary.

Effects of single and multiple transplantations of human umbilical cord mesenchymal stem cells on the recovery of ovarian function in the treatment of premature ovarian failure in mice

BackgroundCurrently, there is no effective treatment for premature ovarian failure (POF), and stem cell therapy is considered the most promising treatment. Human umbilical cord blood mesenchymal stem cells (hUC-MSCs) have shown good regenerative ability in various diseases, including POF; however, their underlying mechanism and dosage for POF treatment remain unclear. This study aimed to compare the effect of single and multiple injections of hUC-MSCs on ovarian function repair in chemotherapy-induced POF.MethodsFemale mice were intraperitoneally injected with 30 mg/kg busulfan and 120 mg/kg cyclophosphamide (CTX) to induce POF. In the single hUC-MSC injection group, hUC-MSCs were transplanted into mice D7 after CTX and busulfan administration, while in the multiple injection group, hUC-MSCs were transplanted on D7, D14, and D21 after CTX and busulfan administration. We evaluated the ovarian morphology, fertility, follicle-stimulating hormone and estradiol concentrations, follicle count, POF model, and cell transplantation results. In addition, real-time polymerase chain reaction, immunohistochemistry, and miRNA and mRNA chips were used to evaluate the effect of the cell therapy.ResultsOvary size, number of follicle at all developmental stages, and fertility were significantly reduced in the POF group compared with the control. Under hUC-MSC treatment, the ovarian morphology and follicle count were significantly restored, and fertility was significantly increased. By comparing the single and multiple hUC-MSC injection groups, we found that the anti-Müllerian hormone and Ki-67 levels were significantly increased in the multiple hUC-MSC group on D60 after chemotherapy. The expression of stimulating hormone receptors, inhibin α, and inhibin β was significantly restored, and the therapeutic effect was superior to that of the single hUC-MSC injection group.ConclusionThese results indicate that hUC-MSCs can restore the structure of injured ovarian tissue and its function in chemotherapy-induced POF mice and ameliorate fertility. Multiple hUC-MSC transplantations have a better effect on the recovery of ovarian function than single hUC-MSC transplantation in POF.

ANTI-MULLERIAN HORMONE AND BASAL ANTRAL FOLLICLE COUNT AS PREDICTORS FOR OVARIAN RESERVE AND OVARIAN RESPONSE IN WOMEN WITH HYPOGONADOTROPIC HYPOGONADISM

To evaluate the use of anti-mullerian hormone (AMH) and basal antral follicle count (AFC) as a predictor of ovarian reserve and ovarian response to controlled ovarian hyperstimulation (COH) in women with hypogonadotropic hypogonadism (HH). This is a retrospective cohort study consisted of the files of 40 women attended a university IVF center during the past 4 years. Nineteen patients has been diagnosed as HH, compared to 21 patients diagnosed as premature ovarian failure (POF). We evaluated the use of AMH value and basal AFC to predict the ovarian reserve and ovarian response to COH in HH women; women with POF have been involved as a control group. We reviewed the recorded AMH and basal AFC, and the total number of retrieved oocytes and fertilized ones in both groups. We used unpaired student t-test for statistical comparison. P<0.05 was considered for statistical significance. Both groups were similar in age, BMI, AMH and basal AFC. Basal FSH and LH were higher in the POF group. Despite the low AMH and the low basal AFC in women with HH ( average 0.46 and 1.6 respectively), we report significant high number of retrieved oocytes and fertilized ones in response to high dose of gonadotropins stimulation compared to the reverse in the POF group. (Table 1) Pregnancy has been reported in 9 women (out of 19) in the HH group. Only one woman (out of 21) got pregnant in the POF group.Table 1Baseline characters and response to COH in women with HH Vs. POF.HH (n=19)POF (n=21)P valueAge26.3323.550.15BMI22.724.90.08FSH0.8724.9<0.001LH0.7521.5<0.001AMH0.460.290.11AFC1.60.910.34Total gonadotrophin dose560040810.03Peak E23344.4427.3<0.001Total retrieved oocytes12.81.3<0.001Total fertilized oocytes8.10.45<0.001 Open table in a new tab AMH and basal AFC has limited value in women with HH regarding prediction of ovarian reserve and ovarian response to COH. Higher doses of gonadotropins can be used rather than early giving up.

Quercetin and flavonoids from Cuscuta chinensis lam. inhibit tripterygium glycoside-induced premature ovarian failure progression via PI3K-AKT signaling pathway

Objective: In this study, we aimed to explore the effects of quercetin and flavonoids extracted from Cuscuta chinensis Lam. on tripterygium glycoside (TG)-induced premature ovarian failure (POF). Materials and Methods: Rats in the POF model were administered with 17β-estradiol (E2), quercetin, or flavonoids that were extracted from C. chinensis. Serum levels of anti-Mullerian hormone (AMH), luteinizing hormone (LH), follicular-stimulating hormone (FSH), and E2 were determined via enzyme-linked immunosorbent assay. The mRNA and protein expressions of PI3K-AKT signaling pathway- and apoptosis-related genes in the ovarian tissues were determined by immunohistochemistry, real-time polymerase chain reaction and western blot analysis. Results: After the administration of E2, quercetin, and flavonoids, there was a decrease in the estrus cycle, which was accompanied by an increase in weight and ovarian indices. TG reduced the thickness of the layer of granulosa cells in the antral follicles, the ratio of primitive follicles, secondary follicles, and sinusoidal follicles, and the number of cells in the corpus luteum and increased the ratio of atresia follicles; however, E2, quercetin, and flavonoids reversed these effects. In the POF group, serum levels of E2 and AMH were decreased, whereas FSH and FSH/LH levels were increased. TG downregulated the expression of PI3K, p-AKT, Atg5, and cyclin D2 and upregulated the expression of caspase-3. However, the changes in the levels of hormones and proteins were reversed after the administration of E2, quercetin, and flavonoids. Conclusion: Quercetin and flavonoids extracted from C. chinensis Lam. inhibited TG-induced POF by modulating the PI3K/AKT signaling pathway.

Human Umbilical Cord Mesenchymal Stem Cells Improve Ovarian Function in Chemotherapy-Induced Premature Ovarian Failure Mice Through Inhibiting Apoptosis and Inflammation via a Paracrine Mechanism

Human umbilical cord mesenchymal stem cell (UC-MSC) application is a promising arising therapy for the treatment of premature ovarian failure (POF). However, little is known about the inflammation regulatory effects of human umbilical cord MSCs (UC-MSCs) on chemotherapy-induced ovarian damage, regardless of in vivo or in vitro. This study was designed to investigate the therapeutic effects of UC-MSC transplantation and underlying mechanisms regarding both apoptosis and inflammation in POF mice. The chemotherapy-induced POF models were induced by intraperitoneal injection of cyclophosphamide. Ovarian function parameters, granulosa cell (GC) apoptosis, and inflammation were examined. Morphological staining showed that UC-MSC treatment increased the ovary size, and the numbers of primary and secondary follicles, but decreased the number of atretic follicles. Estradiol levels in the UC-MSC-treated group were increased while follicle-stimulating hormone levels were reduced compared to those in the POF group. UC-MSCs inhibited cyclophosphamide-induced GC apoptosis and inflammation. Meanwhile, phosphorylation of AKT and P38 was elevated after UC-MSC treatment. Tracking of UC-MSCs in vivo indicated that transplanted UC-MSCs were only located in the interstitium of ovaries rather than in follicles. Importantly, UC-MSC-derived extracellular vesicles protected GCs from alkylating agent-induced apoptosis and inflammation in vitro. Our results suggest that UC-MSC transplantation can reduce ovary injury and improve ovarian function in chemotherapy-induced POF mice through anti-apoptotic and anti-inflammatory effects via a paracrine mechanism.

The Predictive Value of Osteocalcin, Granulin, Cathepsin K and Some Other Biomarkers in Women with Premature Ovarian Failure

Background: Premature ovarian failure (POF), often and misleadingly referred to as ‘premature menopause’,is defined as a loss of ovarian activity before the age of 40 years and is characterized by irregular or absentperiods and reduced fertility. Symptoms include those associated with the natural menopause (night sweatsand vaginal dryness), and with the long-term adverse effects of estrogen deficiency (osteoporosis andcardiovascular disease): the latter is believed to explain the shorter life expectancy associated with POF.Aim: To determinethe predictive value of serum osteocalcin(OC), granulin(GRN), cathepsin k (CTK),vitamin D, parathyroid hormones (PTH), calcium and phosphrus as biomarkers for POF.Method: Sixty (60) women with idiopathic POF with thirty (30) women as control groups were included inthe study. Baseline investigation in all subjects included serum Osteocalcin(OC), granulin(GRN), cathepsink (CTK), vitamin D, parathyroid hormones (PTH), calcium and phosphrus levels were estimated using theappropriate assays for each.Results: The mean serum levels of Osteocalcin, Ganulin,Parathyroid hormones,vitamin D and phosphorus,were significantly higher in women with POF,when compared to healthy controls (P=&lt;0.05). However, meanserum levels of cathepsin k and calcium in POF group were non significant differences,when compared tohealthy controls.Conclusion: Osteocalcin and granulin serum levels may be used as new biomarker for the diagnosis ofpremature ovarian failure.

Discovery of novel serum metabolic biomarkers in patients with polycystic ovarian syndrome and premature ovarian failure

ABSTRACT Several widely recognized metabolites play a role in regulating the pathophysiological processes of various disorders. Nonetheless, the lack of effective biomarkers for the early diagnosis of polycystic ovarian syndrome (PCOS) and premature ovarian failure (POF) has led to the discovery of serum-based metabolic biomarkers for these disorders. We aimed to identify various differentially expressed metabolites (DEMs) through serum-based metabolic profiling in patients with PCOS and POF and in healthy individuals by using liquid chromatography–mass spectrometry analysis. Furthermore, heatmap clustering, correlation, and Z-score analyses were performed to identify the top DEMs. Kyoto Encyclopedia of Genes and Genomes enriched pathways of DEMs were determined using metabolite-based databases. Moreover, the clinical significance of these DEMs was evaluated on the basis of area under the receiver operating characteristic curve. Significantly dysregulated expressions of several metabolites were observed in the intergroup comparisons of the PCOS, POF, and healthy control groups. Furthermore, 6 DEMs were most frequently observed among the three groups. The expressions of these DEMs were not only directly correlated but also exhibited potential significance in patients with PCOS and POF. Novel metabolites with up/downregulated expressions can be discovered in patients with PCOS and POF using serum-based metabolomics; these metabolites show good diagnostic performance and can act as effective biomarkers for the early detection of PCOS and POF. Furthermore, these metabolites might be involved in the pathophysiological mechanisms of PCOS and POF via interplay with corresponding genes.

Curcumin and Hesperidin alleviates oxidative stress and hormonal alterations in a rat model of cyclophosphamide-induced premature ovarian failure

Premature ovarian failure (POF) is a cessation of ovarian function in women under 40 years of age. Ovarian atrophy leads to a reduced follicle reserve, which leads to menstrual irregularities, ovarian dysfunction, and infertility. The beneficial effects of curcumin (CRC) and hesperidin (HSP) on Cyclophosphamide –induced Premature ovarian failure (POF) in a rat model were studied. POF was induced by intraperitoneal (i.p) injection of cyclophosphamide (200 mg/kg b.wt) at day one, and then (8 mg/kg b.wt/day) for the following 14 days. Two weeks after POF induction, treatment with CRC (100 mg/kg b.wt/day, i.p) and HSP (80 mg/kg b.wt/day, i.p) were started and continued for 14 days. Ninety female rats were divided into six equal groups. Group 1( Control normal group) rats received no drugs, Group 2 (POF-induced group), Group 3 (POF + CRC treated), Group 4 ( POF + HSP treated), Group5 ( POF + CRC+HSP treated) and Group 6 (Normal+CRC+HSP treated). Serum follicle-stimulating hormone (FSH) and ovarian tissues Malonedialdehyde (MDA) concentrations were significantly increased while serum estradiol (E2) level, ovarian superoxide dismutase (SOD) activity and reduced glutathione (GSH) concentration were markedly decreased in POF group as compared with control group. However, a significant increase in serum E2 , ovarian tissues SOD activity and GSH level and marked decrease in FSH and MDA concentrations were observed in rats treated with CRC or/and HSP compared with the POF group. Conclusively, curcumin and hesperidin provided an effective treatment for Premature ovarian failure induced by Cyclophosphamide in rats.