Changes In Ovarian Tissue Research Articles

Histopathological Changes in Ovarian Tissues of Rats Underwent Ovarian Ischemia-Reperfusion Injury

This study investigates histopathological changes in ovarian tissues of rats subjected to ischemia-reperfusion (I/R) injury, a condition where restored blood flow causes cellular stress and potential damage. Rats were divided into three groups: control, ischemia, and ischemia-reperfusion. Histological analysis revealed that ischemic ovaries exhibited increased follicular degeneration, atretic follicles, and inflammatory infiltration. The ischemia-reperfusion group showed even more pronounced damage, including disrupted cellular structures and severe vascular congestion. These findings highlight the exacerbated tissue damage caused by reperfusion, underscoring the complex impact of I/R injury on ovarian tissue integrity. Keywords: ovary, ischemia, reperfusion, histology

Loureirin B Reduces Insulin Resistance and Chronic Inflammation in a Rat Model of Polycystic Ovary Syndrome by Upregulating GPR120 and Activating the LKB1/AMPK Signaling Pathway.

Polycystic ovary yndrome (PCOS) is a common metabolic disorder in women, which is usually associated with insulin resistance (IR) and chronic inflammation. Loureirin B (LrB) can effectively improve insulin resistance and alleviate chronic inflammation, and in order to investigate the therapeutic effect of LrB on polycystic ovary syndrome with insulin resistance (PCOS-IR), we conducted animal experiments. A PCOS-IR rat model was established by feeding a high-fat diet combined with letrozole (1 mg/kg·d for 21 days). The rats were treated with the GPR120 agonists TUG-891 and LrB for 4 weeks. Biochemical parameters (fasting blood glucose, total cholesterol, triglycerides, high- and low-density lipoprotein), hormone levels (serum insulin, E2, T, LH, and FSH), and inflammatory cytokines (TNF-α, IL-1β, IL-6, and IL-18) were analyzed. Histopathological analyses of ovaries were performed using hematoxylin/eosin (H&E) staining. Real-time PCR and western blotting were used to assess GPR120, NLRP3, and caspase-1 expression in ovaries, and immunohistochemistry was used to evaluate LKB1 and AMPK protein expression. LrB reduced body weight, Lee's index, ovarian index, ovarian area, and volume in PCOS-IR rats. It lowered fasting blood glucose, serum insulin, and HOMA-IR. LrB decreased total serum cholesterol, triglyceride, and LDL levels and increased HDL levels. It reduced serum T, LH, and LH/FSH and raised serum E2 and FSH levels. LrB downregulated the mRNA and protein expression levels of NLRP3 and Caspase-1, increased the protein and mRNA expression levels of GPR120 in rat ovaries, and increased LKB1 and AMPK protein expression in ovaries, ameliorating ovarian histopathological changes in PCOS-IR rats. Taken together, LrB upregulated GPR120, LKB1, and AMPK protein expression, downregulated NLRP3 and Caspase-1 protein expression, reduced insulin resistance and chronic inflammation, and ameliorated histopathological changes in ovarian tissues in PCOS rats, suggesting its potential as a treatment for PCOS.

Enhancing angiogenesis and inhibiting apoptosis: evaluating the therapeutic efficacy of bone marrow mesenchymal stem cell-derived exosomes in a DHEA-induced PCOS mouse model

BackgroundPolycystic Ovary Syndrome (PCOS) is a widespread endocrine disorder among women, characterized by symptoms like ovarian cysts, hormonal imbalance, and metabolic issues. This research evaluates the therapeutic potential of Bone Marrow Mesenchymal Stem Cell-derived exosomes (BMSC-Exo) in treating PCOS symptoms within a mouse model.MethodsBMSC-Exo were isolated from NMRI mice, characterized using Transmission Electron Microscopy (TEM) and Nanoparticle Tracking Analysis (NTA), and administered to a PCOS mouse model induced by dehydroepiandrosterone (DHEA). The efficacy of BMSC-Exo was assessed in three groups of mice: a control group, a PCOS group, and a PCOS group treated with intravenous BMSC-Exo. Morphological changes in ovarian tissue were examined by Hematoxylin and Eosin (H&E) staining, apoptosis was determined using the TUNEL assay, and CD31 expression was analyzed through immunofluorescent staining to assess angiogenic activity.ResultsThe existence of BMSCs-Exo was confirmed via TEM and NTA, revealing their distinct cup-shaped morphology and a size range of 30 to 150 nanometers. H&E staining revealed that BMSCs-Exo treatment improved ovarian morphology in PCOS models, increasing corpora lutea and revitalizing granulosa cell layers, suggesting a reversal of PCOS-induced damage. TUNEL assays showed that BMSCs-Exo treatment significantly reduced apoptosis in PCOS-affected ovarian cells to levels comparable with the control group, highlighting its role in mitigating PCOS-induced cellular apoptosis. Immunofluorescence for CD31 indicated that BMSCs-Exo treatment normalized endothelial marker expression and angiogenic activity in PCOS models, suggesting its effectiveness in modulating the vascular irregularities of PCOS. Collectively, these findings demonstrate the therapeutic potential of BMSCs-Exo in addressing ovarian dysfunction, cellular apoptosis, and aberrant angiogenesis associated with PCOS.ConclusionThe study substantiates the role of BMSC-Exo in mitigating the deleterious effects of PCOS on ovarian tissue, with implications for enhanced follicular development and reduced cellular stress. The modulation of CD31 by BMSC-Exo further highlights their potential in normalizing PCOS-induced vascular anomalies. These findings propel the need for clinical investigations to explore BMSC-Exo as a promising therapeutic avenue for PCOS management.

Dietary strategies for reducing symptoms associated with endometriosis: a literature review

Endometriosis is an inflammatory pathology characterized by changes in ovarian tissue in women, of reproductive age. People with this disease present symptoms ranging from dysmenorrhea, dyspareunia, chronic pelvic pain, chronic fatigue and difficulty getting pregnant. Therefore, this study is a bibliographical review of the literature with the objective of evaluating the role of nutrition in the treatment of women diagnosed with endometriosis. Thus, the health descriptors (DeCs) used in English and Portuguese were endometriosis, woman and diet using the databases (PubMed, VHL and SciELO), using as subsidies the articles available in full, in English and Portuguese and published in the last 10 years (2013 to 2023). Finally, based on the findings, it is possible to point out the importance of adhering to a healthy diet and the relevant role of micronutrient supplementation, such as vitamins C, D, E and B12 and the minerals Zinc and Potassium, for women with endometriosis.

Inhibition of p38 MAPK/NF-κB p65 signaling pathway activity by rare ginsenosides ameliorates cyclophosphamide-induced premature ovarian failure and KGN cell injury

Ethnopharmacological relevancePanax ginseng C. A. Mey., one of the most used herbs in the world, shows effective treatment in reproductive injury. Recent studies have proven that the processed product, red ginseng, which is more active than ginseng itself. Therefore, it is speculated that its main functional component, rare ginsenosides (heat-transformed saponin, HTS), may be effective in treating premature ovarian failure (POF), but its efficacy has not yet been experimentally confirmed. Aim of the studyTo evaluate whether HTS could attenuate cyclophosphamide-induced inflammation and oxidative damage in POF model rats and the human granulosa-like KGN cell line and protect granulosa cell proliferation. Material and methodsHTS were isolated from ginsenosides and high performance liquid chromatography (HPLC) analysis was used to analyze the HTS components. Cyclophosphamide (CP) was used to establish a POF rat model and KGN cell injury model. Reactive oxygen species (ROS) and antioxidant enzyme production was determined using specific assays, while inflammatory cytokine secretion was measured by enzyme-linked immunosorbent assay (ELISA). The proliferative function of granulosa cells was assessed using high-content screening and immunohistochemistry to determine the Ki67 protein level. Protein expression in ovarian tissues and KGN cells was analyzed by Western blotting, quantitative real-time PCR (qRT-PCR) was used to determine the transcriptional changes in ovarian tissues and KGN cells. ResultsIn CP-treated POF model rats, HTS significantly decreased malondialdehyde (MDA), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) levels, increased glutathione oxidase (GSH) levels, and upregulated Ki67 expression in ovarian granulosa cells. In addition, HTS significantly increased cell survival and Ki67 expression levels in CP-treated cells, and superoxide dismutase (SOD) levels were significantly increased. HTS significantly downregulated IL-6, TNF-α, and interleukin-1β (IL-1β) mRNA expression and significantly inhibited nuclear factor kappa-B p65 (NF-κB p65) and p38 mitogen activated protein kinase (p38 MAPK) phosphorylation in POF model rats and KGN cells. Moreover, NF-κB p65 and p38 MAPK levels were significantly increased in ovarian granulosa cells. p65 and p38 protein and gene expression was significantly downregulated. ConclusionHTS ameliorated CP-induced POF and human granulosa cell injury, possibly by inhibiting inflammation and oxidative damage mediated by the p38 MAPK/NF-κB p65 signaling pathway.

Ginger's Antiapoptotic and Antioxidant Effects on Ovaries of Cyclophosphamide-therapied Rats.

In the recent decade, there has been increasing interest in preventing ovarian toxicity after chemotherapy exposure. It has been documented that ginger (Zingiber officinale) might normalize the hormonal balance and control the menstrual cycle.. This study has analyzed whether ginger extract protects against cyclophosphamide (CP)-induced ovarian failure in rats. Rats were distributed into four groups consisting of vehicle, CP, ginger, and CP + ginger. At the end of the treatment, all rats were killed under anesthesia to obtain ovarian tissues and blood samples for histological, molecular, and biochemical experiments. Our results indicated that ginger improves CP-caused histological changes in ovarian tissues and significantly restores serum hormonal abnormalities. Ginger also showed unique antioxidant, anti-inflammatory, and antiapoptotic properties in the ovarian tissues of CP-induced rats. Further, our findings indicated that ginger might activate the Nrf2 and SIRT and inhibit the PI3K/AKT pathway in the ovaries of CP-treated rats. In conclusion, ginger was found to protect against CP-caused ovarian toxicity in rats. The protective impacts of ginger may mediate, at least partly, by alleviating the oxidant state, inhibiting pro-inflammatory conditions, and exhibiting antiapoptotic activities.

Zigui-Yichong-Fang protects against cyclophosphamide-induced premature ovarian insufficiency via the SIRT1/Foxo3a pathway

Ethnopharmacological relevanceZigui-Yichong-Fang (ZGYCF) is a traditional Chinese medicine prescription for the treatment of infertility and premature ovarian insufficiency (POI). It is clinically used to regulate hormone levels, improve ovarian reserve and increase pregnancy rate. However, the exact mechanism of action is not yet clear. Aims of the studyThis study aimed to explore the potential impact and mechanism of ZGYCF on POI, and provide a scientific basis for its clinical application. Materials and methodsUHPLC‒MS/MS was used to identify the main compounds of ZGYCF. Female 8-week-old C57BL/6N mice were randomized into four group containing the vehicle control (Veh) group, the cyclophosphamide (CTX) model group, the low-dose ZGYCF (CTX-ZG-L) group and the high-dose ZGYCF (CTX-ZG-H) group. A mouse POI model was induced with a single intraperitoneal injection of CTX, and the therapeutic effects of different doses of ZGYCF on POI were evaluated according to the ovarian weight coefficient, serum AMH, serum E2, ovarian histomorphology and follicle counts. After the dose screening experiment, the CTX-ZG-L group was renamed the CTX-ZG group and subjected to follow-up experiments. RNA-seq was used to explore the mechanism of POI and the therapeutic mechanism of ZGYCF on POI in Veh group, CTX group and CTX-ZG group. The mechanism of action of ZGYCF on POI were determined by measuring serum hormone level, histomorphology, follicle counts, protein expression and acetylation modification in groups of Veh, CTX, CTX-ZG and CTX-ZG-Nam (SIRT1 inhibitor). ResultsA total of 37 compounds in ZGYCF were identified. ZGYCF attenuated the morphological changes in ovarian tissue in POI model mice, increased serum AMH and E2 levels, reduced the damage to primordial follicles and other follicles at all stages, and protected ovarian reserve. RNA-seq results suggested that the genes expression of the PI3K signaling and apoptosis signaling pathways was increased in POI mice, while ZGYCF upregulated SIRT1 gene and the expression of estradiol, apoptosis inhibition and other signaling pathway genes. Immunohistochemical staining, TUNEL staining, Western blot analysis and immunoprecipitation results showed that in CTX group, SIRT1 expression and Foxo3a nuclei localization were decreased, while Ac-Foxo3a, p-AKT, p-Foxo3a and apoptotic markers were upregulated. After administration of ZGYCF, these conditions were reversed, however, after treatment with the SIRT1 inhibitor, the results were opposite to those of ZGYCF. ConclusionsAcetylated Foxo3a plays an important role in the occurrence of POI. ZGYCF improves the ovarian reserve of CTX-induced POI mice by activating SIRT1-mediated deacetylation of Foxo3a, and played a role in the treatment of POI. SIRT1 may be a novel target for ZGYCF to ameliorate POI.

Investigation of He's Yang Chao recipe against oxidative stress-related mitophagy and pyroptosis to improve ovarian function.

Primary ovarian insufficiency (POI) is a common gynecological disease with serious ramifications including low pregnancy rate and low estrogen symptoms. Traditional Chinese medicine is regarded as an effective treatment for POI. However, the therapeutic mechanism of it is unclear. In this study, a mouse model of primary ovarian insufficiency was established by intraperitoneal injection of cyclophosphamide (CTX) and He's Yang Chao Recipe (HSYC) concentrate was used for intragastric administration. Serum hormone levels (Anti-Müllerian Hormone, Estradiol, Progesterone, Luteinizing Hormone and Follicle Stimulating Hormone) and Oxidative Stress (OS) related products, superoxide dismutase (SOD), GSH-Px, and malondialdehyde (MDA) were measured by enzyme-linked immunosorbent assay. Pathological changes in ovarian tissue were evaluated by hematoxylin and eosin staining, and flow cytometry was used to determine reactive oxygen species content and mitochondrial membrane potential levels in granulosa cells. Mitochondrial distribution and morphology were investigated using immunofluorescence staining. The level of mitophagy was evaluated by LC3 immunofluorescence staining and autophagosome counts using electron microscopy. Western blotting and qPCR were used to detect the expression of proteins and genes related to mitophagy and the NLRP3 inflammasome. After HSYC treatment, the ovarian damage was milder than in the CTX group. Compared with the CTX group; SOD, GSH-Px, and the total antioxidant capacity were significantly increased, while MDA and ROS were decreased in the HSYC treatment groups. Furthermore, mitochondrial distribution and membrane potential levels were improved after HSYC treatment compared to the CTX group. After the HSYC treatment, the LC3 fluorescent intensity and autophagosome counts were decreased. Similarly, mitophagy related markers PINK1, Parkin, LC3, and Beclin1 were decreased, while p62 was significantly increased, compared with the CTX groups. The mRNA and protein expression of NLRP3 inflammasome, NLRP3, caspase-1, GSDMD, IL-18, and IL-1β were significantly decreased in the HSYC treatment groups. This is the first study in molecular mechanisms underlying HSYC against granulosa cell injury in POI. HSYC protects ovaries from CTX-induced ovarian damage and oxidative stress. HSYC enhanced ovarian function in mice with primary ovarian insufficiency by inhibiting PINK1-Parkin mitophagy and NLRP3 inflammasome activation.

Identification of risk for ovarian disease enhanced by BPB or BPAF exposure

The ban on bisphenol A (BPA) has led to a rapid increase in the use of BPA analogs, and they are increasingly being detected in the natural environment and biological organisms. Studies have pointed out that BPA analogs can lead to adverse health outcomes. However, their interference with ovarian tissue has not been fully elucidated. In this study, seven- to eight-week-old CD-1 mice were exposed to corn oil containing 300 μg/kg/day bisphenol B (BPB) or bisphenol AF (BPAF) through oral gavage, and ovarian tissues were collected at 14 and 28 days of exposure. Ovarian toxicity was evaluated by the ovarian index, ovarian area, and follicle number. mRNA-seq was used to identify differentially expressed genes (DEGs) and infer the association of DEGs with ovarian diseases. BPB or BPAF exposure induced morphological changes in ovarian tissue in CD-1 mice. In addition, Gene Ontology (GO) analysis revealed disturbances in biological processes (BP) associated with steroid biosynthetic process (GO:0006694) and cellular calcium ion homeostasis (GO:0006874). Subsequently, regulatory networks of BPA analogs (BPB or BPAF)-DEGs-ovarian diseases were constructed. Importantly, the expression levels of DEGs and transcription factors (TFs) associated with ovarian disease were altered. BPB or BPAF exposure causes damage to ovarian morphology through the synergistic effects of multiple biological processes and may be associated with altered mRNA expression profiles as a risk factor for ovarian diseases.

Naringenin regulates gut microbiota and SIRT1/ PGC-1ɑ signaling pathway in rats with letrozole-induced polycystic ovary syndrome

PurposeTo evaluate the effect of naringenin on improving PCOS and explore the mechanism. MethodsFirstly, we carried out differential gene expression analysis from transcriptome sequencing data of human oocyte to screen the KEGG pathway, then the PCOS-like rat model was induced by letrozole. They were randomly divided into four groups: Normal group (N), PCOS group (P), Diane-35 group (D), and Naringenin group (Nar). The changes of estrus cycle, body weight, ovarian function, serum hormone levels, glucose metabolism, along with the expression of SIRT1, PGC-1ɑ, claudin-1 and occludin of the ovary and colon were investigated. Furthermore, the composition of the gut microbiome of fecal was tested. ResultsBy searching the KEGG pathway in target genes, we found that at least 15 KEGG pathways are significantly enriched in the ovarian function, such as AMPK signaling pathway, insulin secretion, and ovarian steroidogenesis. Interestingly, naringenin supplementation significantly reduced body weight, ameliorated hormone levels, improved insulin resistance, and mitigated pathological changes in ovarian tissue, up-regulated the expression of PGC-1ɑ, SIRT1, occludin and claudin-1 in colon. In addition, we also found that the abundance of Prevotella and Gemella was down-regulated, while the abundance of Butyricimonas, Lachnospira, Parabacteroides, Butyricicoccus, Streptococcus, Coprococcus was up-regulated. ConclusionOur data suggest that naringenin exerts a treatment PCOS effect, which may be related to the modulation of the gut microbiota and SIRT1/PGC-1ɑ signaling pathway. Our research may provide a new perspective for the treatment of PCOS and related diseases.

MicroRNA-29c-3p participates in insulin function to modulate polycystic ovary syndrome via targeting Forkhead box O 3

ABSTRACT MicroRNAs (miRNAs) are gene expression regulators and changes in miRNA levels are associated with diabetes, insulin resistance, and inflammation, the latter two of which are characteristic of polycystic ovary syndrome (PCOS). The purpose of this study was to explore the specific mechanism in which miR-29 c-3p participated in insulin function to regulate PCOS by targeting Forkhead box O 3 (Foxo3). Peripheral blood from PCOS patients and healthy volunteers were first collected, and the expression levels of miR-29 c-3p and Foxo3 were detected by reverse transcription quantitative polymerase chain reaction or Western blot. Then human granular tumor cell line (KGN) was treated with insulin, and transfected with plasmid vectors interfering with miR-29 c-3p or Foxo3 expression. Cell proliferation was detected by Cell counting kit-8 and plate cloning, and cell apoptosis was tested by flow cytometry. In addition, PCOS rat model was established. PCOS rats were injected with plasmids vectors interfering with miR-29 c-3p or Foxo3 expression, respectively. Pathological changes in ovarian tissues of rats in each group were observed by hematoxylin-eosin staining, and serum sex hormones and glucose metabolism-related indicators were detected. Finally, via bioinformatics website, luciferase digestion report assay was detected the targeting relationship between miR-29 c-3p and Foxo3. The experimental results showed that miR-29 c-3p was down-regulated in PCOS, but Foxo3 was up-regulated. Up-regulated miR-29 c-3p or down-regulated Foxo3 promoted KGN cell proliferation, inhibited apoptosis in vitro, restored PCOS rat sex hormone levels and improved glucose metabolism in vivo. These results suggest that miR-29 c-3p is involved in insulin function to improve PCOS by targeting Foxo3.

Effect of resveratrol on endocrine function, oocyte oxidation and apoptosis in mice with polycystic ovary syndrome


 
 
 
 Purpose: To investigate the effect of resveratrol on endocrine function, oocyte oxidative stress and oocyte apoptosis in mice with polycystic ovary syndrome (PCOS).
 Methods: Healthy female Kunming mice (n = 180) were assigned to control, model and resveratrol groups, respectively. Changes in ovarian tissue in each group, including apoptosis of granulosa cells, were measured. Oocyte abnormal mitochondria, changes in oocyte mitochondrial membrane potential, expression levels of Bcl-2, Bax and caspase-9 in oocytes evaluated. The activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD), and levels of reactive oxygen species (ROS) in each group were also assayed.
 Results: Compared with the model group, the levels of sex hormones, ovarian cell apoptosis and abnormal mitochondria of mice in the resveratrol group were significantly reduced, while uterine weight increased (p < 0.05). The expression levels of Bax, Bcl-2, caspase-9, GPx and SOD, and ROS levels in the resveratrol group decreased significantly (p < 0.05).
 Conclusion: Resveratrol significantly improves endocrine function in PCOS mice, but inhibits oocyte apoptosis and oxidative stress response. Thus, resveratrol may be useful in the treatment of PCOS but further clinical trials are required to confirm it.
 
 
 

Quantification of residual cryoprotectants and cytotoxicity in thawed bovine ovarian tissues after slow freezing or vitrification.

How much residual cryoprotectant remains in thawed/warmed ovarian tissues after slow freezing or vitrification? After thawing/warming, at least 60 min of diffusion washing in media was necessary to significantly reduce the residual cryoprotectants in ovarian tissues frozen by slow freezing or vitrification. Ovarian tissue cryopreservation (OTC) by slow freezing has been the conventional method; while the vitrification method has gained popularity for its practicality. The main concern about vitrification is how much potentially toxic residual cryoprotectant remains in the warmed tissues at the time of transplantation. This was an animal study using the ovarian tissues from 20 bovine ovaries. The duration of this study was from 2018 to 2020. Ovarian cortex tissues were prepared from 20 bovine ovaries and assigned randomly to groups of fresh (non-frozen) control, slow freezing with 1.5 M dimethyl sulfoxide (DMSO), 1.5 M 1,2-propanediol (PROH) and vitrification with 35% ethylene glycol (EG). The residual cryoprotectant concentrations in thawed/warmed tissues were measured by gas chromatography at the following time points: frozen (before thawing/warming), 0 min (immediately after thawing/warming), 30, 60 and 120 min after diffusion washing in media. Next, the ultrastructural changes of primordial follicles, granulosa cells, organelles and stromal cells in the ovarian tissues (1 mm × 1 mm × 1 mm) were examined in fresh (non-frozen) control, slow freezing with DMSO or PROH and vitrification with EG groups. Real-time quantitative PCR was carried out to examine the expressions of poly (ADP-ribose) polymerase-1 (PARP1), a DNA damage sensor and caspase-3 (CASP3), an apoptosis precursor, in thawed/warmed ovarian tissues that were washed for either 0 or 120 min and subsequently in tissues that were ex vivo cultured for 24 or 48 h. The same set of tissues were also used to analyze the protein expressions of gamma H2A histone family member X (γH2AX) for DNA double-strand breaks and activated caspase-3 (AC3) for apoptosis by immunohistochemistry. The residual cryoprotectant concentrations decreased with the extension of diffusion washing time. After 60 min washing, the differences of residual cryoprotectant between DMSO, PROH and EG were negligible (P > 0.05). This washing did not affect the tissue integrity or significantly elevate the percentage of AC3 and γH2AX positive cells, indicating that tissues are safe and of good quality for transplantation. N/A. Since the study was performed with ovarian tissues from bovines, generalizability to humans may be limited. Potential changes in ovarian tissue beyond 120 min were not investigated. This study addresses concerns about the cytotoxicity of EG in warmed ovarian tissues and could provide insights when devising a standard vitrification protocol for OTC. The study was funded by a Grant-in-Aid for Scientific Research (B) from the Japan Society for the Promotion of Science to N.S.

Prognostic value of determination of matrix metalloproteinase-1 and tissue inhibitor of matrix metalloproteinase-1 in follicular fluid in patients in vitro fertilization programs

Aim. To determine the prognostic significance of matrix metalloproteinase-1 (MMP-1) and tissue metalloproteinase-1 inhibitor (TIMP-1) in the follicular fluid (FF) in patients with infertility to improve the effectiveness of in vitro fertilization (IVF) programs.
 Materials and methods. A study of MMP-1 and TIMP-1 in the FF was conducted in 38 patients with infertility in IVF programs. According to the genesis of infertility, the patients were divided into 2 groups. The 1st (control) group included 20 patients with tubal-peritoneal infertility (code N97.1 according to the International Classification of Diseases of the 10th revision ICD-10). The 2nd group (the main one) included 18 patients with tubal-endocrine infertility (ICD-10: N97.1, N97.0, N97.8). The choice of the ovulation stimulation protocol was made taking into account the anamnesis, previously applied protocols, control over the outcome and final result of IVF programs was developed: a protocol with gonadotropin-releasing hormone agonists was used in 20 patients, and a protocol with gonadotropin-releasing hormone antagonists was used in 18 patients. The effectiveness of IVF programs was 26.3%, the frequency of live births was 26.3%. During the puncture of the follicles, FF was taken and the levels of MMP-1 and TIMP-1 were determined in it by the method of quantitative solid-phase enzyme immunoassay of the sandwich type on an automatic flatbed photometer MultiskanFC (2012). The obtained information was subjected to statistical analysis using the Statistica 10.0 application program.
 Results. The level of MMP-1 and TIMP-1 in the FF is an individual indicator and is associated with clinical and anamnestic characteristics. The average level of MMP-1 is 4.9 ng/ml. With negative IVF outcomes, it was 6.11.3 ng/ml, with positive outcomes 1.40.1 ng/ml. The content of MMP-1 in women with tubal-peritoneal infertility genesis of 7.91.7 ng/ml indicates a residual inflammatory process in the follicle. The average level of TIMP-1 was 2462.264.3 ng/ml. The indicator increases: with age (2535.0125.7 ng/ml), duration of infertility (2611.5126.7 ng/ml), due to surgical interventions on the reproductive organs (2800122.7 ng/ml) and in patients with tubal-endocrine genesis of infertility (2550126.7 ng/ml), which indicates fibrosclerotic changes in the follicles and ovarian tissue.
 Conclusion. The level of MMP-1 in the range of 1.31.4 ng/ml should be considered a criterion for the successful onset of pregnancy in IVF programs. The criterion of subclinical damage to the collagen matrix in the FF is the level of MMP-1 6.1 ng/ml or higher; it can be considered as an unfavorable prognostic factor for the outcome of IVF. The level of TIMP-1 in the range of 24002450 ng/ml should be considered a criterion for the successful onset of pregnancy in IVF programs. The probability of a negative outcome of IVF is in the range of 15002300 ng/ml (inability to level the effects of MMP-1 with these indicators), while the level of 2696 ng/ml and higher in tubal-endocrine genesis of infertility reflects a high degree of fibrosclerotic changes in ovarian tissue and can be considered as an unfavorable prognostic factor of IVF outcome.

Nitrite exposure may induce infertility in mice.

In the present study, we investigated the potential of nitrite exposure to induce infertility in mice. Adult female C57BL/6J mice were randomly divided into control and nitrite exposure groups. Subsequently, the rate of mouse infertility was calculated, and pathological changes in ovarian tissues were examined using hematoxylin and eosin staining. In addition, TUNEL staining, immunofluorescent labeling, and western blotting were performed to assess cell apoptosis and oxidative stress response in ovarian tissues from various groups. We observed that nitrite exposure could induce infertility (p<0.05) in mice. High-dose nitrite exposure caused infertility in a time-dependent manner, and two-round exposure induced higher infertility than that one-round exposure (p<0.01). In addition, a higher number of atretic follicles were detected in the ovaries of nitrite-exposed groups than in the control group. Furthermore, TUNEL-positive cells were observed in granulosa cells of atretic follicles, and overexpression of caspase 8, c-Fos, and inducible nitric oxide synthase (iNOS) was detected in ovaries after nitrite exposure (p<0.01), suggesting that cell apoptosis and oxidative stress response were induced following nitrite exposure. Collectively, these findings suggest that nitrite exposure can induce mouse infertility in a time-dependent manner. Oxidative stress response and cell apoptosis are involved in mediating nitrite-induced infertility.

Bu-shen-zhu-yun decoction inhibits granulosa cell apoptosis in rat polycystic ovary syndrome through estrogen receptor α-mediated PI3K/AKT/mTOR pathway

Ethnopharmacological relevanceBu-shen-zhu-yun decoction (BSZYD) is a traditional chinese herbal prescription is widely used in the treatment of infertility. Aim of the studyWe aimed to elucidate the impact of a traditional herbal prescription BSZYD on polycystic ovary syndrome (PCOS). Materials and methodsThe candidate active compounds in BSZYD and their putative targets were investigated by bioinformatics analysis. A deydroepiandrosterone (DHEA)-induced PCOS rat model was then constructed using female Sprague-Dawley (SD) rats. Serum hormone levels were measured by enzyme-linked immunosorbent assay (ELISA). Pathological changes in ovarian tissues were analyzed with hematoxylin and eosin (H&E) staining. The expressions of estrogen receptor α (ER α)-mediated PI3K/AKT/mTOR pathway were measured by immunofluorescence and western blotting. ResultsBioinformatics analysis showed that the putative targets of active compound candidates in BSZYD were enriched in PI3K/AKT and estrogen signaling pathways related to regulating ovarian ovulation. Animal experiments showed that BSZYD significantly alleviated pathological changes in the ovary, altered hormone levels of serum and reduced apoptosis rate of granulosa cells. In addition, BSZYD treatment notably upregulated the expressions of proteins in ER α-mediated PI3K/AKT/mTOR pathway and downregulated apoptosis-related proteins in PCOS rats. ConclusionBSZYD can restore ovary lesions and ameliorate apoptosis through ER α-mediated PI3K/AKT/mTOR pathway, which might partly contribute to the treatment of PCOS.

Efficacy of an Yinyanghuo (Herba Epimedii Brevicornus)-Xianmao (Rhizoma Curculiginis) drug pair in a rat model of polycystic ovary syndrome.

To identify active compounds in an Yinyanghuo (Herba Epimedii Brevicornus) - Xianmao (Rhizoma Curculiginis) drug pair (ECD) and investigate its efficacy on polycystic ovary syndrome (PCOS), and its possible mechanism in a rat model of PCOS. A network pharmacology approach involving a characteristic drug assessment, active compound and target prediction, PCOS gene collection as well as network analysis was employed. The ovary morphology after treatment was observed using an animal model and western blotting and real-time PCR were used to verify AKT1 as the molecular target. Six networks were constructed, an active compound-target network for the ECD (C-T network), a drug-target network (D-T network), a related genes network, a targets interaction network, a key genes interaction network, and a gene-pathway network. A total of 41 compounds and 261 targets were identified for the ECD, 232 PCOS-related genes, 31 cogenes, and 14 pathways. These pathways may be involved in the efficacy of ECD on PCOS. The proteins most involved in the signal pathways for all targets were AKT1, IL6, INSR, ESR, and GSK3B. The AKT1 target was selected for experimental verification. Based on the Western blot and real-time PCR results, the expression of AKT1 in the PCOS model varied after treatment with ECD. Our findings suggest that the ECD can reverse the negative morphological changes in ovarian tissue that occur in model rats of PCOS. AKT1 may be a key mediator of the observed ability of the ECD to protect against PCOS in the model rats.

Dynamics of changes in the main indicators of reproductive health of women receiving chemotherapy for malignant breast tumors

Breast cancer is the second most common cancer in the world and the most common cancer among women. The use of chemotherapy as a component of comprehensive treatment leads to the development of chemoinduced premature ovarian failure. The aim of the study was to examine the dynamics of changes in the main indicators of reproductive health of women who received chemotherapy for malignant neoplasms of the breast. Reproductive health of patients (levels of antimullerian hormone, estradiol and follicle-stimulating hormone, ovarian volume and number of antral follicles) before chemotherapy and one, two, and three years after completion of chemotherapy was studied according to the purpose and objectives of the study. The obtained data were processed using the statistical software package SPSS 20.0 for Windows. Quite significant changes in the reproductive health of women in the study group were manifested in the form of menstrual disorders (oligomenorrhea, amenorrhea), involuntary abortions, inability to conceive. Changes in hormonal status were characterized by a progressive decrease in antimullerian hormone and estradiol levels and an increase in follicle-stimulating hormone levels. In addition, there was a decrease in ovarian volume and a decrease in the number of antral follicles. The dynamics of changes in the studied indicators was characterized by a sharp deterioration during the first year after chemotherapy, followed by less rapid progression. This trend is explained by dystrophic changes in ovarian tissue after chemotherapy, and at a later date – in addition, the age-related involution of the ovaries. The dynamics of changes in the main hormonal and ultrasound parameters of the ovarian reserve indicates significant impairment of reproductive function in women receiving chemotherapy for malignant neoplasms of the breast.

Quercetin attenuates cisplatin-induced ovarian toxicity in rats: Emphasis on anti-oxidant, anti-inflammatory and anti-apoptotic activities

Ovarian toxicity is a devastating adverse effect of cisplatin therapy. The objective of the current study was to address whether or not quercetin could protect against cisplatin-induced ovarian toxicity in rats as well as the possible underlying mechanisms. Rats were allocated into five groups. Group 1 represented the control, group 2 was administered quercetin (10 mg/kg), group 3 received cisplatin (6 mg/kg single i.p. dose) on days 7 and 14, the forth group was given cisplatin + quercetin (5 mg/kg) and the fifth group was administered cisplatin + quercetin (10 mg/kg). Quercetin ameliorated cisplatin-induced histopathological changes in ovarian tissues and significantly prevented the decline in the percentage of healthy follicles and serum anti-Mullerian hormone (AMH). Quercetin exhibited significant anti-oxidant effects evidenced by preventing MDA accumulation, glutathione depletion and superoxide and glutathione peroxidase exhaustion in the ovary. Also, quercetin displayed activities against cisplatin-induced inflammatory responses in the ovary. Quercetin significantly inhibited expression of NFκb, Cox-2 and IL-6 and elevated ovarian content of TNF-α. Further, quercetin showed anti-apoptotic activity as demonstrated by decreased caspase-3 content and modulation of Bax and Bcl2 expression in the ovary. Conclusively, quercetin protects against cisplatin-induced ovarian toxicity in rats. This is mediated, at least partly, by its anti-oxidant, anti-inflammatory and anti-apoptotic activities.

Possible protective effects of crocin on destructive side effects of cyclo-phosphamide in mice ovarian tissue: Evaluation of histomorphometrical and biochemical changes.

One of the side effects of cyclophosphamide (CP) is low fertility. In this study, we investigated the protective role of crocin (Cr) against CP chemotherapy-induced changes in ovarian tissue. In the current study, we treated 15 female mice aged 6-8 weeks old for 21 days. The mice were distributed into three groups including control received normal saline (0.10 mL; IP), CP or sham-control group (CP once a week, 15.00 mg kg-1; IP) and experimental (CP + Cr) group received CP along with Cr (200 mg kg-1 daily; IP). After completing the procedure, levels of total anti-oxidant capacity (TAC), superoxide dismutase (SOD) and sex hormones in serum as well as malondialdehyde (MDA) in the left ovarian tissue were measured. The right ovaries were used for histological and morphological tests. The obtained data were statistically analyzed by SPSS software using ANOVA and Tukey follow-up studies. Results showed that in the CP group a significant decrease was observed in ovarian follicles, the number of corpus luteum, levels of TAC, SOD and sex hormones; while, there was a significant increase in the number of atretic follicles and mast cells and level of MDA compared to control group. Administration of Cr along with CP caused a significant ameliorative effect on the studied parameters. In conclusion, the Cr could significantly decrease the side effects caused by CP chemotherapy in mice ovarian tissue.