Effects of lupeol and flutamide on experimentally-induced polycystic ovary syndrome in mice.

Objectives: Polycystic ovary syndrome (PCOS) is one of the main causes of infertility in women. This study was conducted to uncover the effects of lupeol as an anti-androgenic triterpene on experimentally-induced PCOS in mice. Materials and Methods: Eighty immature female mice were divided into 4 groups: Control (C), PCOS (P), Lupeol (L), and Flutamide (F). PCOS was induced in test groups by injection of Dehydroepiandrosterone (60 mg/kg/day, IP) for twenty days. Following the PCOS induction, the two groups of L and F were treated with lupeol (40 mg/kg/day) and/or flutamide (10 mg/kg/day) respectively and the two groups of C and P received sesame oil (0.1 ml/mouse/day) for 15 days. After the treatment period, ten animals in each group were selected for collecting blood and ovary samples. In vitro fertilization assessment was carried out on 10 remaining mice in each group. The hormonal assays and oxidative stress biomarker determination were performed on serum and tissue samples. Moreover, histopathological analyses were conducted on the ovaries. Results: PCOS-elevated concentration of LH and Testosterone was significantly (P<0.05) lowered in lupeol and flutamide-received animals. Lupeol and flutamide both reduced PCOS-induced fibrosis and the number of atretic follicles. Both compounds declined the PCOS-increased lipid peroxidation and protein oxidation in serum and the ovaries. Lupeol increased the PCOS-reduced fertility rate and decreased the number of arrested embryos by 12%. Conclusion: These findings indicate that lupeol could be a novel compound in the treatment of PCOS as it reduced PCOS-induced structural and also functional disorders.

Profiling and Functional Analysis of long non-coding RNAs in yak healthy and atretic follicles.

Yak is the livestock on which people live in plateau areas, but its fecundity is low. Follicular development plays a decisive role in yak reproductive performance. As an important regulatory factor, the expression of long non-coding RNA (lncRNAs) in yak follicular development and its regulatory mechanism remains unclear. To explore the differentially expressed lncRNAs between healthy and atretic follicular in yaks. We used RNA-seq to construct lncRNA, miRNA, and mRNA expression profiles in yak atretic and healthy follicles, and the RNA sequence results were identified by qPCR. In addition, the correlation of lncRNA and targeted mRNA was also analyzed by Starbase software. Moreover, lncRNA/miRNA/mRNA networks were constructed by Cytoscape software, and the network was verified by dual-luciferase analysis. A total of 682 novel lncRNAs, 259 bta-miRNAs, and 1704 mRNAs were identified as differentially expressed between healthy and atretic follicles. Among them, 135 mRNAs were positively correlated with lncRNA expression and 97 were negatively correlated, which may be involved in the yak follicular development. In addition, pathway enrichment analysis of differentially expressed lncRNA host genes by Kyoto Genome Encyclopedia (KEGG) showed that host genes were mainly involved in hormone secretion, granulosa cell apoptosis, and follicular development. In conclusion, we identified a series of novel lncRNAs, constructed the lncRNA ceRNA regulatory network, and provided comprehensive resources for exploring the role of lncRNAs in yak ovarian follicular development.

Atresia of ovarian follicles in fishes, and implications and uses in aquaculture and fisheries.

Atresia of ovarian follicles, that is the degenerative process of germ cells and their associated somatic cells, is a complex process involving apoptosis, autophagy and heterophagy. Follicular atresia is a normal component of fish oogenesis and it is observed throughout the ovarian cycle, although it is more frequent in regressing ovaries during the postspawning period. An increased occurrence of follicular atresia above physiological rates reduces fish fecundity and even causes reproductive failure in both wild and captive-reared fish stocks, and hence, this phenomenon has a wide range of implications in applied sciences such as fisheries and aquaculture. The present article reviews the available literature on both basic and applied traits of oocyte loss by atresia, including its morpho-physiological aspects and factors that cause a supraphysiological increase of follicular atresia. Finally, the review presents the use of early follicular atresia identification in the selection process of induced spawning in aquaculture and the implications of follicular atresia in fisheries management.

Unraveling the mechanism of long-term bisphenol S exposure disrupted ovarian lipids metabolism, oocytes maturation, and offspring development of zebrafish.

Bisphenol S (BPS) acts as a xenoestrogen and disturbs the female reproductive system; however, the underlying mechanism has not been elucidated. In this study, the effect of chronic BPS exposure (1 µg/L and 100 µg/L) on ovarian lipid metabolism in zebrafish was investigated to determine its influence on adult reproductive capacity and offspring development. The results showed that long-term (240 days) exposure to BPS induced lipid accumulation in the ovaries by promoting the transport of more lipids from the circulation to the ovaries and by upregulating triacylglycerol synthesis-related genes. Significantly increased expression of cpt2, acadm, acadl, and pparα, which are involved in ß-oxidation in the ovarian mitochondria, indicated that more energy was provided for oocyte maturation in exposed zebrafish ovaries. Thus, the proportion of full-grown stage oocytes in ovaries and egg reproduction were elevated at an accelerated rate, which earlier than normal reproductive cycle (8-10 days posts pawning). Moreover, the maternally BPS-exposed F1 embryos (2 h post-spawning, hpf) showed higher neutral lipid levels, impaired hatching capacity, and increased occurrence of larval deformities. All these findings demonstrated that stimulated lipid synthesis and ß-oxidation in zebrafish ovaries significantly contribute to BPS-induced oocyte precociousness with subsequent effects on the development of unexposed offspring. This study provides new insight into the impact of xenoestrogens on oviparous reproduction in females and offspring development from the perspective of ovarian lipid metabolism.

Novel extra cellular-like matrices to improve human ovarian grafting.

PURPOSE: To investigate if human ovarian grafting with pure virgin human recombinant collagen type-1 from bioengineered plant lines (CollPlant™) or small intestine submucosa (SIS) yields better implantation results for human ovarian tissue and which method benefits more when combined with the host melatonin treatment and graft incubation with biological glue + vitamin E + vascular endothelial growth factor-A. METHODS: Human ovarian tissue wrapped in CollPlant or SIS was transplanted into immunodeficient mice with/without host/graft treatment. The tissue was assessed by follicle counts (including atretic), for apoptosis evaluation by terminal deoxynucleotidyl transferase assay and for immunohistochemical evaluation of neovascularization by platelet endothelial cell adhesion molecule (PECAM) expression, and for identification of proliferating granulosa cells by Ki67 expression. RESULTS: Human ovarian tissue transplanted with CollPlant or SIS fused with the surrounding tissue and promoted neovascularization. In general, implantation with CollPlant even without additives promoted better results than with SIS: significantly higher number of recovered follicles, significantly fewer atretic follicles, and significantly more granulosa cell proliferation. Moreover, results with CollPlant alone seemed to be at least as good as those after host and graft treatments. CONCLUSIONS: CollPlant is a biomaterial without any potential risks, and grafting ovarian tissue with CollPlant is easy and the procedure may be easily modified, with limited or no foreseeable risks, for auto-transplantation in cancer survivors. Further studies are needed using other novel methods capable of enhancing neovascularization and reducing apoptosis and follicle atresia.

Ameliorative effects of quercetin on folliculogenesis in diabetic mice: a stereological study.

A high risk of reproductive disorders can be seen in diabetic pregnancy. Reproductive disorders associated with diabetes may result from alterations in the function of the ovary. In this study, we investigated the ameliorative effects of quercetin as a phytoestrogen and antidiabetic agent on the folliculogenesis in diabetic mice. Streptozotocin-induced diabetic mice were treated with 30 mg/kg/day quercetin for four weeks. The volume of ovary, follicles, and corpus luteum were significantly decreased in the diabetic mice. The number of growing follicles (secondary, antral, and Graafian follicles) and corpus luteum was significantly decreased in the diabetic mice. Also, the volume of oocytes was significantly decreased in antral and Graafian follicles. Our results indicated that the administration of quercetin in diabetic mice increased the volume of the ovary and growing follicles, the number of growing follicles and corpus luteum. It also significantly decreased the number of atretic follicles. As a result, it may be concluded that the impaired follicular growth and development caused by hyperglycemia in diabetic mice can be alleviated by quercetin treatment.

Morphologic analysis of atretic follicles in the Chinese soft-shelled turtle, Pelodiscus sinensis.

The Chinese soft-shelled turtle (Pelodiscus sinensis) and its eggs have been utilized as both a source of nutrition and medicine for thousands of years. In the present study, follicular atresia was investigated by morphological analysis as an important factor affecting clutch size. Results showed that follicular atresia can be divided into four stages: (1) development of follicular cell disorder; (2) massive cellular proliferation in the ooplasm with elimination of yolk granules; (3) large amounts of erythrocytes appearing in the ooplasm; (4) gradual degeneration of atretic follicles. For the first time, we have shown follicular atresia to be a dynamic process in P. sinensis and found that yolk granules are eliminated by both liquefaction as well as non-liquefaction processes. The present study may provide detailed morphologic analysis for further research into egg quality and clutch size in P. sinensis.

Histological Changes of the Ovary of Adult Mice Exposed Prenatally to Flunitrazepam / Cambios Histológicos del Ovario de Ratones Adultos Expuestos Prenatalmente a Flunitrazepam

Flunitrazepam (FNZ) is a minor tranquillizer involving allosteric modulation of the GABA receptor complex. It is a ligand of the peripheral benzodiazepine receptor (PBR) that participates in cholesterol transport in steroidogenic organs. The purpose was to investigate whether a single oral dose of FNZ of 2.5 mg/kg of body weight (bw), administered on day 6 of gestation, alters the structure of the adult ovary of mouse offsprings at 2 months of age. The mouse offsprings of the in utero FNZ-treated group and those of the control group (C) were killed. Ovaries were obtained in early estrous, fixed in Zenker solution, and processed by routine histological techniques. Serial sections were observed under light microscopy to determine the characteristics and quantity of follicles in different stages of development and of the corpus luteum. The ovarian tissues from the FNZ group depicted a great staining affinity, enlarged nuclei with abundant heterochromatin clumps. The quantity of primordial, primary and secondary normal follicles in the FNZ group decreased significantly (p< 0.01). The number of primary atretic follicles increased (p<0.01) and the secondary ones remained constant as in group C. Histological changes and statistical data suggest that FNZ produces long-lasting epigenetic or genotoxic effects in follicular and corpus luteum cells of the ovary of prenatally FNZ-treated mice.

El flunitrazepam (FNZ) es un tranquilizante menor que actúa modulando el receptor GABA, es un ligando del receptor benzodiazepínico periférico (PBR), el cual participa en el transporte de colesterol en órganos esteroidogénicos. El objetivo de este trabajo fue investigar si una dosis oral de FNZ (2.5mg/kg de peso) administrada en el sexto día de gestación, altera la estructura del ovario en crías de ratón adultos. Las crías del grupo FNZ tratadas in utero con la benzodiazepina, así como el grupo control, fueron sacrificadas. Los ovarios se extrajeron en estro temprano, se fijaron en líquido de Zenker y procesaron con las técnicas habituales de histología. Los cortes seriados fueron analizados mediante microscopia óptica, los folículos fueron identificados y contados de acuerdo a los diferentes estadios del desarrollo, así como las células del cuerpo lúteo. El tejido ovárico en el grupo de FNZ presentó una gran afinidad tintórea, núcleos grandes y con abundantes cúmulos de heterocromatina. El número de folículos primordiales, primarios y secundarios en el grupo FNZ disminuyó (p<0.01). Contrario al número de folículos atrésicos que aumentaron (p<0.01), a excepción de los folículos atrésicos secundarios que no fueron diferentes a los controles. Las alteraciones histológicas y los datos estadísticos sugieren que el FNZ produce un efecto epigenético o genotóxico de largo plazo tanto en los folículos como en las células del cuerpo lúteo, de ratones tratados prenatalmente con el fármaco.