Follicles Of Rats Research Articles

Brown adipose tissue activation with ginsenoside compound K ameliorates polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is a common metabolic and endocrine disease affecting women of reproductive age. Due to its complex aetiology, there is no currently effective cure for PCOS. Brown adipose tissue (BAT) activity is significantly decreased in PCOS patients, and BAT activation has beneficial effects in animal models of PCOS. Here, we investigated the effect of ginsenoside compound K (CK) in an animal model of PCOS and its mechanism of BAT activation. Primary brown adipocytes, Db/Db mice and dehydroepiandrosterone (DHEA)-induced PCOS rats were used. The core body temperature, oxygen consumption, energy metabolism related gene and protein expression were assessed to identify the effect of CK on overall energy metabolism. Oestrous cycle, serum sex hormone, ovarian steroidogenic enzyme gene expression and ovarian morphology were also evaluated following CK treatment. Our results indicated that CK treatment could significantly protect against body weight gain in Db/Db mice via BAT activation. Furthermore, we found that CK treatment could normalize hyperandrogenism, oestrous cyclicity, normalize steroidogenic enzyme expression and decrease the number of cystic follicles in PCOS rats. Interestingly, as a potential endocrine intermediate, C-X-C motif chemokine ligand-14 protein (CXCL14) was significantly up-regulated following CK administration. In addition, exogenous CXC14 supplementation was found to reverse DHEA-induced PCOS in a phenotypically similar manner to CK treatment. In summary, CK treatment significantly activates BAT, increases CXCL14 expression and ameliorates PCOS. These findings suggest that CK might be a potential drug candidate for PCOS treatment.

An investigation of the effects of chronic zonisamide, sultiam, lacosamide, clobazam, and rufinamide anti-seizure medications on foliculogenesis in ovarian tissue in prepubertal non-epileptic rats.

We aimed to determine the morphological and histological effects of zonisamide, sultiam, lacosamide, clobazam, and rufinamide on ovarian folliculogenesis in rats. Sixty female Wistar rats were divided into six experimental groups as control, zonisamide, sultiam, lacosamide, clobazam, and rufinamide groups; control solution and drugs were administered by gavage for 90 days. The number of healthy follicles in the control group was significantly higher than in the anti-medication groups (p< 0.001), and the number of corpus luteum was significantly lower (p< 0.001). There was a significant difference in the number of TUNEL positive apoptotic follicles between the control and drug groups (p< 0.001). With EGF, IGF-1, and GDF-9 staining, a very strong immunoreaction was observed in the ovarian multilaminar primary follicle granulosa cells and oocytes in the control group compared to the drug group (p< 0.001). Long-term anti-seizure medication with zonisamide, sultiam, lacosamide, clobazam, and rufinamide from prepubertal to adulthood causes apoptosis and disruption of folliculogenesis in the ovarian follicles of nonepileptic rats.

Androgen Excess Induced Mitochondrial Abnormality in Ovarian Granulosa Cells in a Rat Model of Polycystic Ovary Syndrome.

BackgroundAndrogen excess could profoundly lead to follicular dysplasia or atresia, and finally result in polycystic ovary syndrome (PCOS); however, the exact mechanism remains to be fully elucidated.MethodsPCOS model rats were induced by dehydroepiandrosterone, and their fertility was assessed. The ovarian granulosa cells (GCs) from matured follicles of PCOS model rats were collected and identified by immunofluorescence. The mitochondrial ultrastructure was observed by transmission electron microscope and the mitochondrial function was determined by detecting the adenosine triphosphate (ATP) content and mtDNA copy number. Besides, the expressions of respiratory chain complexes and ATP synthases in relation to mitochondrial function were analyzed.ResultsThe PCOS model rats were successfully induced, and their reproductive outcomes were obviously adverse. The GCs layer of the ovarian was apparently cut down and the mitochondrial ultrastructure of ovarian GCs was distinctly destroyed. The ATP content and mtDNA copy number of ovarian GCs in PCOS model rats were greatly reduced, and the expressions of NDUFB8 and ATP5j were significantly down-regulated without obvious deletion of mtDNA 4834-bp.ConclusionsAndrogen excess could damage mitochondrial ultrastructure and function of GCs in rat ovary by down-regulating expression of NDUFB8 and ATP5j in PCOS.

Bunium incrassatum Bois. Batt. Trab. (Talghouda) in the improvement of thyroid tissue damages in female rats

This study aims to determine the nutritional and therapeutic effects of dried Bunium incrassatum Bois.Batt.Trab. Tubers powder on rats with hyper and hypothyroidism. Thirty female Wistar rats were divided into 6 groups. G1 (C) received a normal diet. G2 (CT) received a normal diet with dried Talghouda tubers powder dissolved in water. G3 (Hpo) was treated with Carbimazole® (5 mg/day) for 6 weeks. G4 (HpoT) was treated with Carbimazole® (5 mg/day) for 6 weeks and Talghouda for 7 weeks after induced hypothyroidism. G5 (Hpy) was treated with Levothyroxine® (600µg/Kg body weight/day) for 6 weeks. G6 (HpyT) had hyperthyroidism induced by Levothyroxine® (600µg/Kg body weight/day) for 6 weeks and then treated with Talghouda for 7 weeks. A dose of Talghouda was chosen at 1.03g/Kg body weight/day, and all treatments were given by gavage. Phytochemical analysis was carried out for Talghouda, and the thyroid glands of each group were then recovered for histological study. Talghouda treatment showed a highly significant (p 0.01) increase in body weight in groups G2, G4 and G6 with gland repair and reactivation of thyroid follicles in hyper and hypothyroid rats compared to G1, G3 and G5. The powder of dried Talghouda tubers, used as traditional medicine in Algeria, improved the damages caused by hyper and hypothyroidism.

Influence of Diabetes on Morphometric Index of Ovarian Follicles in Streptozocin-Induced Rats

The study aimed to determine the effect of diabetes on follicle development by measuring the diameters of rats' ovarian follicles. The structure and function of many organs change by being affected by diabetes. The ovary is an essential organ of the reproductive system affected by diabetes. The size of the ovarian follicles and corpus luteum can also be affected by diabetes. For this reason, body mass, blood glucose level, and rat ovarian follicles, and corpus luteum diameters were measured in this study. Ten healthy female rats were kept as the control group. In the other rat group, experimental diabetes was induced with streptozotocin (STZ)(60 mg/kg). The rats in both groups were killed after 30 days and their ovaries removed. 5-6 µm sections were made using paraffin embedding techniques and stained with hematoxylin-eosin. On the fifth day of STZ administration to rats, the mass loss of rats was 10%, and the diabetogenic index was 330%. Compared with the control group, diameters of diabetic rats significantly decreased the diameter of the primordial, primary and Graafian follicles, and corpus luteum. The decrease in the diameter of secondary follicles of diabetic rats was not significant. The percentage shrinkage index was the highest in the corpus luteum with 37%. As a result, it can be said that diabetes influences the size of the ovarian follicles and especially the corpus luteum, thereby negatively affecting the ability to improve oocyte quality. Diabetes-related follicle diameter may shrink and cause infertility. It may be essential to measure the diameters of the follicles in vitro fertilization studies in patients with diabetes.

Hypothyroidism and Fertility: An Animal Model follows up in The Second-Generation.

Objective Hypothyroidism is known as the most common endocrine disorder. The prevalence of hypothyroidism in the female and male population is 2% and 0.2%, respectively. Maternal hypothyroidism is a defect in the thyroid hormones transition from the mother to the fetus. The present study was conducted to find whether maternal hypothyroidism affects the fertility of the second generation. Materials and Methods In this experimental study, twelve adult female rats weighting 180-220 g were randomly divided into case and control groups. Hypothyroidism was induced by dissolving 0.1 g/L of 6-n-propyl-2-thiouracil in drinking water toward the end of pregnancy and lactation. At the end of the breastfeeding period, the blood samples of female children were collected. Six healthy, mature, female rats were selected and kept until they reached maturity, and were then mated with male rats. After observing the female rats’ delivery, blood samples were collected from their male and female newborns and the healthy rats were selected. Results There was a significant difference in the volume and size of ovarian as well as in the number of secondary follicles in comparison with the control group (P=0.025). However, there were no significant changes in the other parameters including the number of primary follicles, the number of Graafian follicles and sperm parameters. There was no significant decrease in the testicular volume and size, number of Leydig cells and seminiferous tubules diameter. ConclusionMaternal hypothyroidism has no significant effects on testicular tissue function, and sperm parameters in the second generation, but can significantly reduce the rate of secondary follicles in the second generation female rats.

Capsaicin prevents radiotherapy-induced premature ovarian failure in rats.

Ionising radiation exposure of 5-10gray (Gy) to the pelvic area induces premature ovarian failure (POF). Twenty-four young adult Wistar albino female rats were were treated with subcutaneous capsaicin 0.5mg/kg per day or placebo for 10days then exposed to whole body irradiation. Rats were randomly divided into four groups: (1) control; (2) capsaicin; (3) radiation only (IR): rats were injected with placebo before exposure to a single dose of 8.3-Gy whole body irradiation; (4) radiation-capsaicin (IR+CAP): rats were injected with capsaicin prior to whole body irradiation. Radiation triggered oxidative stress, increased ovarian inflammation, increased follicular apoptosis and diminished ovarian follicle pool. Capsaicin significantly ameliorated oxidative stress by decreasing serum total oxidant status, oxidative stress index, disulphide, and malondialdehyde levels (P ≤0.001); ovarian inflammatory status by decreasing expressions of TNF-α, IL-1β, PARP-1 (P =0.002); apoptosis by decreasing expressions of active caspase-3 and p53 (P =0.015, P =0.002); and follicle counts by increasing primordial follicles and decreasing apoptotic follicles (P ≤0.001) in rats when administered before radiation exposure. The beneficial effects of capsaicin are demonstrated for the first time on ionising radiation exposed rat ovaries. Capsaicin pre-treatment before radiotherapy restores the primordial follicle pool, inhibits atresia of ovarian follicles and may be an acceptable therapeutic modality to prevent radiation-induced POF.

CaCO3-based carriers with prolonged release properties for antifungal drug delivery to hair follicles.

Superficial fungal infections are of serious concern worldwide due to their morbidity and increasing distribution across the globe in this era of growing antimicrobial resistance. The delivery of antifungals to the target regions of the skin and sustaining the effective drug concentration are essential for successful treatment of such mycoses. Topical formulations get extra benefits here if they penetrate into the hair follicles since fungal hyphae can proliferate and produce spores in such reservoirs. We designed a novel particulate system for the encapsulation and intrafollicular delivery of griseofulvin (Gf) antifungal drug, which is water-insoluble and currently commercially available in oral dosage forms. Micron-sized calcium carbonate (vaterite) carriers containing 25 ± 3% (w/w) of Gf were prepared via the wet chemical method. The successful in vivo transportation of the carriers into the hair follicles of rats was demonstrated using scanning electron and confocal laser scanning microscopy. In addition, we introduced an approach toward Gf release prolongation for the proposed system. The stabilizing coatings were formed on the surface of the obtained particles via the layer-by-layer technique. The formulations displayed sufficient biocompatibility and good cellular uptake in contact with fibroblast cells in vitro. Four different coatings were tested for their preserving ability in the course of continued carrier incubation in the model media. The best release prolonging formulation liberated 38% of the loaded Gf during 5 days, while the uncoated carriers demonstrated more than 50% drug release within the first 24 h in water. To assess the in vivo release properties, free Gf drug and Gf-loaded carriers (uncovered and covered with the stabilizing shell) were administered topically in rats and the drug excretion profiles were further studied. By comparing the daily Gf levels in urine, we verified the sustained effect (longer than a week) of the stabilizing shell formed on the carrier surface. Conversely, the application of the free drug did not provide reliable Gf detection for this period. These findings open new prospects for the efficiency enhancement of topical therapeutics. Importantly, the elaborated system could be adapted for the dermal delivery of various water-insoluble drugs beyond the scope of antifungal therapy.

Bushen Jieyu Tiaochong Formula reduces apoptosis of granulosa cells via the PERK-ATF4-CHOP signaling pathway in a rat model of polycystic ovary syndrome with chronic stress

Ethnopharmacological relevancePolycystic ovary syndrome (PCOS) is a common and complex endocrine disorder that is also an important cause of infertility. Adverse psychological stress can aggravate the occurrence and development of PCOS. Bushen Jieyu Tiaochong Formula (BJTF), a prescription of Traditional Chinese Medicine (TCM), has been used in the treatment of PCOS and shown to be effective in reducing negative emotion. However, the therapeutic mechanism has yet to be clearly elucidated. In the current study, we investigated the potential mechanism of action of BJTF. Aim of the studyTo investigate the role of PERK-ATF4-CHOP signaling in the molecular mechanisms that mediate the effects of BJTF in a rat model of PCOS, with chronic stress induced by letrozole and a chronic unpredictable mild stress (CUMS) paradigm. Materials and methodsIn addition to the normal control group, the PCOS combined with CUMS model rats were randomly assigned to a model group, a Diane-35 (ethinylestradiol 35 μg/cyproterone acetate 2 mg)-treated positive control group, or one of three BJTF-treated groups receiving a low, medium, or high dose. Behavioral testing, including the sucrose preference test and open field test, was conducted, and hematoxylin and eosin (H&E) staining was used to observe changes in the pathological morphology of ovarian tissue. Free testosterone (FT), luteinizing hormone (LH), and follicle-stimulating hormone (FSH) levels in serum were quantified by enzyme-linked immunosorbent assays (ELISA). The hippocampal levels of norepinephrine (NE), 5-hydroxytryptamine/serotonin (5-HT), and 5-hydroxyindoleacetic acid (5-HIAA) were measured using high-performance liquid chromatography–electrochemical detection (HPLC-ECD). Apoptotic granulosa cells were detected using terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Furthermore, quantitative reverse transcription polymerase chain reaction (qRT-PCR) and immunohistochemistry were used to detect the expression of glucose-regulated protein 78 (GRP78) and CHOP in the ovarian tissues. The expression levels of GRP78, CHOP, PERK, and ATF4 in ovarian tissues were also measured by western blotting. ResultsTreatment with either BJTF or Diane-35 ameliorated the abnormal cystic dilatation of follicles in the model rats and reduced the serum levels of FT and LH, and the LH/FSH ratio. BJTF treatment also attenuated chronic psychological stress-like behavior and regulated the expression and metabolism of cerebral monoamine neurotransmitters. The efficacy of BJTF was greater than that of Diane-35, with the optimal effects observed at the medium dose. BJTF also lowered the apoptotic index of ovarian granulosa cells and downregulated the expression of GRP78, CHOP, and ATF4. Although the expression level of PERK was not significantly altered by BJTF, the mean PERK expression level was the lowest in the medium-dose BJTF group. ConclusionsAdministration of BJTF has the therapeutic potential to promote the homeostasis of the reproductive endocrine environment and to restore follicular development and ovulation, possibly through the inhibition of the PERK-ATF4-CHOP signaling pathway, leading to downregulation of GRP78 expression to further delay ovarian granule cell apoptosis mediated by endoplasmic reticulum stress (ERS). Moreover, BJTF could improve behavioral performance by regulating cerebral monoamine neurotransmitters in this rat model. These findings provide a new perspective for treating PCOS related to psychological stress using TCM.

ERβ Regulation of Gonadotropin Responses during Folliculogenesis.

Gonadotropins are essential for regulating ovarian development, steroidogenesis, and gametogenesis. While follicle stimulating hormone (FSH) promotes the development of ovarian follicles, luteinizing hormone (LH) regulates preovulatory maturation of oocytes, ovulation, and formation of corpus luteum. Cognate receptors of FSH and LH are G-protein coupled receptors that predominantly signal through cAMP-dependent and cAMP-independent mechanisms that activate protein kinases. Subsequent vital steps in response to gonadotropins are mediated through activation or inhibition of transcription factors required for follicular gene expression. Estrogen receptors, classical ligand-activated transcriptional regulators, play crucial roles in regulating gonadotropin secretion from the hypothalamic–pituitary axis as well as gonadotropin function in the target organs. In this review, we discuss the role of estrogen receptor β (ERβ) regulating gonadotropin response during folliculogenesis. Ovarian follicles in Erβ knockout (ErβKO) mutant female mice and rats cannot develop beyond the antral state, lack oocyte maturation, and fail to ovulate. Theca cells (TCs) in ovarian follicles express LH receptor, whereas granulosa cells (GCs) express both FSH receptor (FSHR) and LH receptor (LHCGR). As oocytes do not express the gonadotropin receptors, the somatic cells play a crucial role during gonadotropin induced oocyte maturation. Somatic cells also express high levels of estrogen receptors; while TCs express ERα and are involved in steroidogenesis, GCs express ERβ and are involved in both steroidogenesis and folliculogenesis. GCs are the primary site of ERβ-regulated gene expression. We observed that a subset of gonadotropin-induced genes in GCs, which are essential for ovarian follicle development, oocyte maturation and ovulation, are dependent on ERβ. Thus, ERβ plays a vital role in regulating the gonadotropin responses in ovary.

Intra-ovarian injection of bone marrow-derived c-Kit+ cells for ovarian rejuvenation in menopausal rats.

Introduction: Cell-based therapies with certain cell types are touted as novel and hopeful therapeutic intervention in the clinical setting. Here, we aimed to assess the regenerative potential of c-Kit+ cells in the rejuvenation of ovarian tissue and fertility rate in rat model of premature ovarian failure (POF). Methods: Rats were treated with 160 mg/kg/BW of 4-vinylcyclohexene dioxide for 15 days. Freshly enriched rat bone marrow-derived c-Kit+ (MACS) and c-Kit- cells (4×105 cells/10 µL) were transplanted into the ovaries of treatment and control animals. Prior to transplantation as well as 2, 4, 6, and 8 weeks post-transplantation, randomly-selected rats were euthanized and ovarian tissues were subjected to pathophysiological examinations and real-time PCR analyses. Results: POF status was confirmed by the presence of pathological features and a decreased number of immature and mature follicles compared with the control group (P < 0.05). Histological examination revealed a substantial reduction of atretic follicles in POF rats receiving c-Kit+ cells in comparison with POF rats that did not receive these cells (P < 0.05). Compared with the control samples, angiogenesis-related genes, Angpt2 and KDR, showed increased and decreased expressions in POF ovaries, respectively (P < 0.05). c-Kit+ cells had potential to restore angiogenesis in the ovarian tissue within normal ranges. Systemic levels of FSH did not significantly change in pre- or post-transplantation time points for any group (P > 0.05). Notable reduction of collagen deposition was found in c-Kit-treated rats. Transplantation of c-Kit+ cells also restored the reduced fertility rate (P < 0.05). Conclusion: The administration of c-Kit+ cells can modulate angiogenesis and pathological changes, leading to the rejuvenation of ovarian function of a rat model of premature menopause.

P–443 Effects of capsaicin pre-treatment on ovarian follicle pool, inflammatory and apoptotic pathways against radiation induced ovarian failure

Abstract Study question Is capsaicin effective in preventing radiation induced ovarian follicle loss and premature ovarian failure (POF) in rats? Summary answer Capsaicin pre-treatment before radiotherapy restores especially primordial follicle pool, inhibits atresia of ovarian follicles, may be an acceptable therapeutic modality to prevent radiation induced POF. What is known already Ionizing radiation exposure to pelvic area induces inflammation, oxidative stress, follicular atresia and apoptosis; leading to POF. Phytochemicals were used in animal studies to prevent radiotherapy induced POF because of their antioxidant and anti-inflammatory properties however their potential radio-protective effects in human ovarian follicles are not clear. Capsaicin is the active compound of hot peppers and has anti-inflammatory and antioxidant properties. It was found that low dose capsaicin stimulated ovarian follicular development and proliferation of granulosa cells, inhibited apoptosis of ovarian follicles in pre-pubertal rat ovaries. However, no data exists on radio-protective effects of capsaicin on ovarian follicles. Study design, size, duration Twenty-four young adult Wistar albino female rats were housed under standard conditions (20 ± 1 0C room temperature, 60 ± 10% humidity, and a 12/12-h light/dark cycle) in regular cages and allowed free access to food and water. After 10 days of subcutaneous capsaicin 0,5 mg/kg/day or placebo treatment, animals exposed to total body irradiation of 8.3 Gy using a linear accelerator. Treatment continued for 1 day after irradiation. Participants/materials, setting, methods Rats were randomly divided into four groups: (1) control: non-irradiated rats were injected placebo; (2) capsaicin: non-irradiated rats were injected capsaicin; (3) radiation only (IR): rats were injected placebo before exposure to a single dose of 8.3-Gy whole body radiation; (4) Radiation-capsaicin (IR+CAP): rats were injected capsaicin prior to whole body irradiation and continued for 1 day after irradiation. Rats were sacrificed, blood samples were obtained for biochemical investigations. Ovaries were dissected for histopathological evaluation. Main results and the role of chance Radiation triggered oxidative stress, increased ovarian inflammation, increased follicular apoptosis and diminished ovarian follicle pool. Capsaicin was significantly ameliorated; oxidative stress by decreasing serum total oxidant status, oxidative stress index, disulfide, and malondialdehyde levels (p ≤ 0.001 both); ovarian inflammatory status by decreasing expressions of TNF-α, IL–1β, poly ADP-ribose polymerase–1 (PARP–1) (p = 0.002 both); apoptosis by decreasing expressions of active caspase–3 and p53 (p = 0.015 and p = 0.002 respectively); follicle counts by increasing primordial follicles and decreasing apoptotic follicles (p ≤ 0.001 both) in rats when administered before radiation exposure. Results of our study confirmed previously reported pro-proliferative and anti-apoptotic properties of capsaicin on ovarian follicles. These beneficial effects of capsaicin are demonstrated for the first time on ionizing radiation exposed rat ovaries. Limitations, reasons for caution Present study is a in-vivo rat study and other preclinical studies are needed to confirm our findings before moving forward to human trials. Radio-protective effects of capsaicin on rat ovarian follicles were demonstrated only in short term. Long term effects of capsaicin on folliculogenesis, fertilization and fecundity should be investigated. Wider implications of the findings: Preserving fertility is one of the main goals of successful radiotherapy in terms of quality of life for oncological or hematological diseases. Capsaicin treatment before radiotherapy may be an acceptable therapeutic modality to prevent radiation induced POF and has potential to utilize in clinical application in terms of fertility preservation. Trial registration number 218S876/2019

Magnolia officinalis Ameliorates Dehydroepiandrosterone-Induced Polycystic Ovary Syndrome in Rats

Background: Polycystic ovary syndrome (PCOS) is a prevalent reproductive and metabolic disorder. Insulin resistance (IR) is highly associated with PCOS and aggravates its symptoms. Thiazolidinediones (TZDs), as insulin sensitizing agents, are PPARγ agonists that improve many of the symptoms of PCOS. The Magnolia officinalis extract (MOE) is a natural peroxisome proliferator activated receptor gamma (PPARγ) agonist that improves insulin sensitivity in experimental models. Objectives: Using a dehydroepiandrosterone (DHEA)-induced rat model of PCOS and IR, this study aimed to explore both the potential beneficial effects and the molecular mechanisms of action of MOE. Methods: Post-pubertal female Sprague Dawley rats were subcutaneously injected daily with DHEA (6 mg/100 g body weight) dissolved in sesame oil for 28 days (n = 30). Age- and weight-matched control rats received only sesame oil (n = 12). Afterward, 16 of the DHEA-injected rats, along with five control rats, were sacrificed for blood and tissue collection. The 14 remaining DHEA-injected rats received either treatment of 30 days of oral MOE (500 mg/kg) dissolved in dimethyl sulfoxide (DMSO) (n = 7), or oral DMSO only (n = 7). Meanwhile, the remaining control rats (n = 7) continued to receive daily oral DMSO for 30 days. At the end of the treatments, the rats were sacrificed for blood and tissue collection. Results: After 28 days, the DHEA-treated rats exhibited an increase in body weight as compared to controls (P &lt; 0.05). DHEA injection induced a PCOS phenotype as evident by a statistically significant (P &lt; 0.05) elevated serum luteinizing hormone (LH), and an increased number of cystically dilated follicles with thicker granulosa compared to controls. PCOS rats showed a statistically significant rise in fasting insulin with an increased homeostatic model assessment index of insulin resistance (HOMA-IR) as compared to controls (P &lt; 0.05). Compared to the control group, PCOS rats had a statistically significant lower ovarian protein expression of PPARγ, insulin receptor substrate 1 (IRS1), and protein kinase B (Akt) by Western Blot (P &lt; 0.05). Conversely, the PCOS group showed an increased mammalian target of rapamycin (mTOR) pathway activity as evident by an increase in the fraction of phosphorylated mTOR to total mTOR compared to the control group (P &lt; 0.05). When treated for 30 days with oral MOE (500 mg/kg), the PCOS rats showed a statistically significant decrease in body weight and serum LH levels as compared to the non-treated PCOS rats (P &lt; 0.05). The number of cystically dilated follicles in the MOE-treated PCOS rats was significantly reduced compared to the non-treated PCOS rats. In the MOE-treated PCOS rats, the ovarian protein expression of PPARγ, IRS1, and Akt was significantly increased, while the p-mTOR/mTOR expression was decreased compared to the non-treated PCOS group (P &lt; 0.05). Conclusions: According to our results, the MOE ameliorated the DHEA-induced PCOS phenotype histologically, hormonally, and metabolically. Fundamentally, this explores the elusive pathophysiologic association between IR and PCOS by targeting pathways common to both disorders.

Effect of Acute Exposure of Cigarette Smoke on Endometrium Thickness and Number of Ovarian Follicles in Rats

Background: Cigarrete smoke has been ascociated with many health-related problems especially in long time use. One of many health problems that are affected is the reproductive system that give negative impact in fertility. However, there are limited studies about this in acute exposure of cigarette smoke. Objective: The aim of this study is to investigate the effect of acute exposure of cigarette smoke on endometrial thickness and number of ovarian follicles in rats. Methods: Thirty adult female rats were randomly divided into 3 groups: P0, P1 and P2. The P1 and P2 groups were exposed to cigarette smoke for 15 days with dose of 2 and 4 cigarettes daily, respectively. Vaginal swab was done twice, before and after the last treatment, to determine the proestrus phase of the animals. Histological slide of uterus and ovarium using Haematoxylin Eosin staining were observed under light microscop. Number of ovarian follicles were measured, and the thickness of endometrium was meassured using CellSens software version 2.3 (Olympus Life Science). Statistical analysis of data was performed using MANOVA in SPSS software version 23 for Windows. Results: There were no significant differences in both endometrial thickness and number of ovarian follicles between 3 groups (p&gt;0.05). However, we found that there were a downward trend of some variables (endometrial thickness, number of primary follicles, de Graff follicles, and corpus luteum) from P0 to P2. Conclusion: We concluded that acute exposure of cigarette smoke not yet influenced the female reproductive organ, based on histological appearance of endemotrium and ovarium.

Histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with metformin during and after induction of permanent estrus

Objective: To evaluate the histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with metformin during and after induction of permanent estrus. Methods: Thirty-two adult-female rats with regular estrous cycle were equally divided into four groups: 1) GCtrl - at estrous phase. 2) GPCOS - at permanent-estrous phase. 3) GMet1 - rats and daily treated with metformin (12.5 mg/Kg) during 60 consecutive days, as preventive form and 4) GMet2 - PCOS rats, which remained exposed to 60 days of continuous illumination and treated with metformin. After that, the animals were euthanized, and the ovaries were removed and processed for paraffin embedding. Sections were stained with H.E. for histomorphometry or subjected to immunohistochemistry for Ki-67 and cleaved caspase-3 (Casp-3) detections. Results: The GPCOS showed lack of corpus luteum and several ovarian cysts, as well as interstitial-like cells. The presence of corpus luteum and a significant increase in primary and antral follicles were observed in Mel-treated groups, which also showed a decrease in the number of ovarian cysts and in the area occupied by interstitial-like cells. The presence of corpus luteum along with an increase in the number of primary follicles in the Met2 group were noticed (p&lt;0,01). A significant reduction in number of cysts and in the area occupied by interstitial cells, as well as a decrease in nuclear volume of interstitial cells, were noticed in the Met-treated groups, mainly in the Met2 group. The percentage of cell proliferation was significantly higher in granulosa cells of the Met-treated groups than PCOS group, mainly in the GMet2 (p&lt;0,01), which was similar to the GCtrl group. On the other hand, the percentage of apoptosis (cleaved-caspase-3- positive cells) was significantly higher in the granulosa cells of GPCOS and Met-treated groups than the GCtrl group, but without significant difference, which showed weak cleaved caspase-3 immunoreactivity in those cells. Conclusion: The ovaries of rats treated with metformin showed a decrease in nuclear volume and in the area occupied by interstitial cells, presence of corpus luteum, in addition to a decrease in the number of cysts.

Улучшение деградации лекарственных носителей на основе ватерита внутри волосяных фолликулов с помощью сонофореза

Intrafollicular drug delivery is beneficial in terms of both localized therapy of relevant skin disorders and systemic transportation of bioactive molecules. Vaterite particles are capable of loading and delivering various substances to hair follicles. Possibility to control the duration of their intrafollicular degradation can improve such a particulate delivery system. Here, we propose the use of sonophoresis (1 MHz, 1 W/cm2 ) to accelerate the resorption of vaterite carriers inside the hair follicles of rats in vivo. The effect of sonication is demonstrated utilizing optical coherence tomography monitoring of the skin and scanning electron microscopy investigation of the plucked hairs. A nine-minute post-treatment of skin in the site of particle delivery allowed us to almost halve the time of their degradation.

Development of a Chemical Reproductive Aging Model in Female Rats.

Women are born with an abundant but finite pool of ovarian follicles, which naturally and progressively decreased during their reproductive years until menstrual periods stop permanently (menopause). Perimenopause represents the transition from reproductive to non-reproductive life. It is usually characterized by neuroendocrine, metabolic and behavioral changes, which result from a follicular depletion and reduced number of ovarian follicles. During this period, around 45-50 years old, women are more likely to express mood disorders, anxiety, irritability and vasomotor symptoms. The current animal models of reproductive aging do not successfully replicate human perimenopause and the gradual changes that occur in this phase. While the traditional rat model of menopause involves ovariectomy or surgical menopause consisting of the rapid and definitive removal of the ovaries resulting in a complete loss of all ovarian hormones, natural or transitional menopause is achieved by the selective loss of ovarian follicles (perimenopause period). However, the natural aging rodent (around 18-24 months) model fails to reach very low estrogen concentrations and overlaps the processes of somatic and reproductive aging. The chronic exposure of young rodents to 4-vinylcyclohexene diepoxide (VCD) is a well-established experimental model for perimenopause and menopause studies. VCD induces loss of ovarian small follicles (primary and primordial) in mice and rats by accelerating the natural process of atresia (apoptosis). The VCD, ovary-intact or accelerated ovarian failure (AOF) model is the experimental model that most closely matches natural human progression to menopause mimicking both hormonal and behavioral changes typically manifested by women in perimenopause. Graphical abstract: The female reproductive system is regulated by a series of neuroendocrine events controlled by central and peripheral components. (A). The mechanisms involved in this control are extremely complex and have not yet been fully clarified. In female mammals whose ovulation (the most important event in a reproductive cycle) occurs spontaneously, reproductive success is achieved through the precise functional and temporal integration of the hypothalamus-pituitary-ovary (HPO) axis. (B). In women, loss of fertility appears to be primarily associated with exhaustion of ovarian follicles, and this process occurs progressively until complete follicular exhaustion marked by the final menstrual period (FMP). (C). While in female rodents, reproductive aging seems to begin as a neuroendocrine process, in which changes in hypothalamic/pituitary function appear independently of follicular atresia. The traditional rat model of menopause, ovariectomy or surgical menopause consists of the rapid and definitive removal of the ovaries resulting in a complete loss of all ovarian hormones. (D). The chronic exposure (15-30 days) to the chemical compound 4-vinylcyclehexene diepoxide (VCD) in young rodents accelerates gradual failure of ovarian function by progressive depletion of primordial and primary follicles, but retains residual ovarian tissue before brain alterations that occurs in women in perimenopause. Low doses of VCD cause the selective destruction of the small preantral follicles of the ovary without affecting other peripheral tissues.

Pemberian Ekstrak Oncom Hitam dan Merah Memperpanjang Siklus Estrus dan Mempertebal Endometrium Tikus Putih

hytoestrogens are active compounds, derived from plants, which have a similar structure and function as estrogen. Phytoestrogens are commonly found in legumes. Oncom, which is assumed containing phytoestrogens, is one of the most famous legumes food from Indonesia and widely consumed daily in West Java. This study was aimed to determine the effect of oncom extract on estrous cycle, endometrium thickness, and the number of antral follicles in productive age rats (Rattus novergicus). This experimental study was using 21 three-to-four-month-old fertile female rats and divided into three groups. Group I (K) was considered as a control group without any treatment. Group II and III were treatment groups which were given black (H) and red (M) oncom extracts 0.005 g/g body weight, respectively, orally with a feeding tube for 14 days. The length of the estrous cycle was measured by performing vaginal swab with interval 12 hours after first treatment was given and during the treatment. Endometrium thickness and the number of antral follicles were measured by collecting the organs uterus and ovary for histological purpose with paraffin method after rats were euthanized post-treatment oncom extracts for 14 days. Data were analyzed by ANOVA and continued with LSD test. The total length of estrous cycle of control group, black oncom extract group, and red oncom extract group was 107,43±3,16 hours, 141,43±15,36 hours, and 161,14±17,10 hours, respectively. The mean of endometrium thickness of control group, black oncom extract group, and red oncom extract group was 346,945±65,88 ?m, 485,740±86,69 ?m, and 533,904±78,93 ?m, respectively. The number of antral follicles of control group, black oncom extract group, and red oncom extract group was 6,00±1,54, 8,43±2,99, and 9,14±2,72, respectively. Results showed that black and red oncom extracts had a significant effect on the length of estrous cycle and endometrium thickness in rats, yet there is no significant difference in the number of antral follicles. In summary, black and red oncom extracts had effects on the length of estrous cycle and endometrium thickness, yet there was no effect on the number of antral follicles.

Ajwa Improves Nicotine Induced Ovarian Toxicity in Murine Model

Introduction: Nicotine, a major component of cigarette smoke, alters ovarian function and Ajwa (Phoenix dactylifera) being an antioxidant prevents its toxicity. Aims &amp; Objectives: This experimental study was designed to evaluate protective effect of Ajwa against ovarian toxicity induced by nicotine in adult albino rats. Place and duration of study: The study was conducted in the Department of Anatomy, Federal Postgraduate Medical Institute, Shaikh Zayed Medical Complex, Lahore for duration of 3 months (January 2017-March 2017). Material &amp; Methods: 40 adult female albino rats were divided into four equal groups. 1ml distilled water was given to the rats of control (A) group by gastric intubation, experimental group B was given nicotine injection intraperitoneally (0.1 mg/kg body weight) for 28 days. The rats of experimental group C were given Ajwa fruit extract 1gm/kg body weight daily by gastric intubation, while rats of experimental group D were given 0.1mg/kg body weight nicotine intraperitoneally plus 1gm/kg body weight Ajwa fruit extract by gastric intubation for 28 days. Results: In group B, secondary and Graafian follicles were mostly atrophic, degeneration of granulosa cells, pyknotic nuclei and stromal vacuolization were present. Zona pellucida was torn and irregular. Mean diameters of secondary and Graafian follicles were significantly lower in nicotine group B (p-value&lt;0-001*). Experimental group C revealed normal histological architecture and diameter of ovarian follicles. Near normal restoration of follicular structure was seen in group D. Conclusion: Ajwa fruit extract is counterbalancing for nicotine induced oxidative stress in ovarian follicles of rat, so it may be useful to improve infertility or sub-fertility induced by smoking in females.

Histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats with polycystic ovaries treated with clomiphene citrate

Objective To evaluate the histomorphometric and immunohistochemical changes in interstitial cells and ovarian follicles of rats treated with clomiphene citrate during and after induction of permanent estrus. Methods Twenty four adult-female rats with regular estrous cycle were equally divided into three groups: (1) GCtrl–at estrous phase. (2) GPCOS–at permanent-estrous phase. (3) GCC–PCOS rats, which remained exposed to 60 days of continuous illumination and treated with Clomiphene Citrate. After that, the animals were euthanized, and the ovaries were removed and processed for paraffin embedding. Sections were stained with H.E. for histomorphometry or subjected to immunohistochemistry for Ki-67 and cleaved caspase-3 detections. Results The GPCOS showed lack of corpus luteum and several ovarian cysts, as well as interstitial-like cells. The presence of corpus luteum and a significant increase in primary and antral follicles were observed in GCC, which also showed a decrease in the number of ovarian cysts and in the area occupied by interstitial-like cells, as well as a decrease in nuclear volume of interstitial cells. The percentage of cell proliferation was significantly higher in granulosa cells of the GCC. On the other hand, the percentage of apoptosis was significantly higher in the granulosa cells of GPCOS than the GCC. Conclusion The ovaries of rats treated with clomiphene citrate showed a decrease in the number of cysts, an increase in the number of ovarian follicles, the presence of corpus luteum along with a decrease in the nuclear volume in the area occupied by interstitial cells.