Cystic Follicles Research Articles

Scanning the Future: Veterinary Reproductive Ultrasonography


 Reproductive ultrasonography can be an extensively preferred diagnostic modality in farm and companion animals for maximizing reproductive health in future days. The shape, contour, size, and position of the structure being analyzed, as well as its echogenicity, which is determined by the amplitude of the echoes received, are being used to describe ultrasound pictures. Reproductive ultrasonography has introduced a new dimension to animal reproduction by allowing not only visualization of the reproductive tract but also early pregnancy diagnosis, surveillance of embryonic or fetus development, and detection of the estrous cycle phase. Confirmation of ovarian and uterine ailments such as cystic ovarian follicle, endometritis, hydrometra, mucometra, and pyometra can be easily diagnosed for the earliest treatment regimens. The technique is a simple, safe, non-invasive, cost-effective, and practical way to confirm reproductive status in farm and companion animals from 30 days of gestation onwards, and by trans-rectal approach, the earliest diagnosis of pregnancy and non-pregnancy in small ruminants and swine can be confirmed from day 20 onwards. As a result, it appears that veterinary reproductive ultrasonography is an excellent technique for managing fertility in farm and companion animals

Astragalus hamosus Acts as an Insulin Sensitizer in the Treatment of Polycystic Ovary Syndrome Rat Models by Affecting IRS1 Expression.

Polycystic ovary syndrome (PCOS) is the most common endocrine abnormality among women of reproductive age. Insulin resistance is known as the hallmark of PCOS that leads to hyperinsulinemia and type 2 diabetes in PCOS patients. This study aimed to evaluate the expression pattern of IRS1 as a candidate gene in insulin resistance development in the PCOS rat models. In this study, estradiol valerate was used for PCOS induction. Then, all of the rats were divided into five experimental groups and treated with Astragalus hamosus extract. Ethanol was used for extraction by Soxhlet, and extracts were analyzed by GC-MS. Ovarian morphology was analyzed using histological experiments. Finally, the expression of IRS1 and hormonal titration of testosterone and insulin were evaluated using qRT-PCR and ELISA assays, respectively. Induction of PCOS led to an increase in body weight, which decreased after treatment with the extract. Histological assessment declared an increased number of corpora lutea in treated groups and reduced cystic follicles compared to the PCOS group. Astragalus hamosus extract-treated groups exhibited decreased levels of insulin and testosterone compared to the PCOS group. qRT-PCR results showed an increase in the expression levels of IRS1 in the treated groups compared to the PCOS group. This study indicated the impact of Astragalus hamosus extract on PCOS by clarifying the increased levels of IRS1 expression in the treated groups compared to the PCOS group.

Alterations of Cortisol and Melatonin Production by the Theca Interna Cells of Porcine Cystic Ovarian Follicles.

Simple SummaryThe mechanism of follicular cyst formation is largely unknown but changes in follicular composition are known to be involved. In particular, there is abnormal hormone secretion in cystic follicles. Here, we found there was disruption of hormone secretion in the fluid of cystic follicles in sows. The glucocorticoid receptor was highly expressed, and the melatonin receptor was weakly expressed in cystic follicles compared with control follicles. Thus, secretion of steroid hormones in cystic follicles is disrupted and disturbances in signaling via cortisol and melatonin are involved in the development of follicular cysts in sows.(1) Background: Cortisol and melatonin (MT) act in regulating follicular development. We hypothesized that abnormal levels of cortisol, MT, and steroids in theca interna cells might be involved in the development of follicular cysts in sows. (2) Methods: To test this hypothesis, we measured the mRNA levels of enzymes involved in steroid hormone synthesis, the glucocorticoid receptor (GR), and melatonin receptors (MTRs) in theca interna cells of cystic and normal porcine follicles. (3) Results: The concentrations of estradiol, progesterone, and cortisol were greater in cystic follicles than in control ones (p = 0.034, p = 0.020, p = 0.000), but the concentration of MT was significantly lower (p = 0.045). The levels of GR, 11β-HSD1, and 11β-HSD2 were higher in cystic follicles than in control l follicles. MT types 1 and 2 were significantly lower in cystic follicles (p < 0.05). The mRNA expression levels of genes encoding the steroid hormone synthesis enzymes, steroidogenic acute regulatory protein (StAR), recombinant cytochrome P45011A1 (CYP11A1), and 3β-hydroxysteroid dehydrogenase (3β-HSD) in theca interna cells of cystic follicles were significantly higher than in control follicles. Thus, there was disruption of hormone secretion in the fluid of cystic follicles in sows. (4) Conclusions: The levels of steroid hormones, cortisol and MT are disrupted in porcine cystic follicles.

Case of Cystic Graffian Follicles in Cattle

Case of Cystic Graffian Follicles in Cattle

Histomorphological Changes in a Rat Model of Polycystic Ovary Syndrome and the Contribution of Stevia Leaf Extract in Modulating the Ovarian Fibrosis, VEGF, and TGF-β Immunoexpressions: Comparison with Metformin.

Polycystic ovary syndrome (PCOS) is a common endocrine disorder of fertile females. It has been reported that stevia leaf extract (SLE) has antidiabetic and antihyperlipidemic properties. Therefore, the current study hypothesized and investigated the role and mechanistic aspects of a natural sweetener; SLE in treating a rat model of letrozole-induced PCOS and to compare it with metformin. Thirty-five female Wistar albino rats were divided into 5 groups: control, PCOS-induced group (letrozole, 1 mg/kg/d, for 21 days), SLE, metformin, and combination-treated groups (300 mg/kg/d, for the next 28 days in SLE and metformin-treated groups). Vaginal smears were done. The levels of glucose, lipid, and hormonal profiles were measured in the serum meanwhile, malonyl dialdehyde (MDA), superoxide dismutase (SOD), and tumour necrosis factor (TNF-α) were measured in the ovary. Ovarian sections were subjected to hematoxylin and eosin, Masson, and immunohistochemical identification of VEGF and TGF-β followed by morphometric analysis. PCOS rats showed altered hormonal and lipid profiles, in addition to hyperglycemia. Also, the ovarian tissue levels of MDA and TNF-α were elevated, and SOD was decreased. Numerous cystic follicles, decrease/absence of corpora lutea, interstitial fibrosis with positive VEGF and TGF-β immunoreactivity were evident. SLE improved all altered parameters. SLE showed potential therapeutic merits in letrozole-induced PCOS via anti-inflammatory, antioxidant, anti-fibrotic, and angiogenesis regulating mechanisms. Its effects were almost comparable to metformin, and the combination of both has no further synergistic effect.

Therapeutic potential of hydrolysable tannin on weight management oxidative stress and reproductive health in polycystic rats

Phytonutrients get attention now days due to their wide application and their tendency to improve the productive and reproductive performance in humans. Hydrolysable tannin (HT) is one of these bionutrients. The present study was aimed to investigate the therapeutic effect of HT on the induced polycystic ovarian syndrome (PCOS) in rats. Forty-five adult female rats with 4 to 5 days regular estrous cycle were selected. The PCOS was induced by a single intramuscular injection of Estradiol Valerate 4 mg/rat/kg. The rats were offered 0.5, 1, 1.5 and 2% of HT/kg body weight through oral gavage for two months. The results had shown a reduction (p<0.05) in average feed intake, body weight and weight gain, total oxidative stress and nutrient digestibility in PCOS rats. Similar reduction trend (p<0.05) in serum luteinizing hormone and insulin was observed; while serum follicular stimulating hormone, serum prolactin and total antioxidant capacity were improved (p<0.05). The ovarian histopathology showed no signs of reduction in cystic follicles while no improvement was observed in healthy follicles. The study concluded that the treatment with 1.5% level of HT had shown the best therapeutic potential for weight management, reduction of oxidative stress and improvement in reproductive health of PCOS rats.

Normotensive rats with PCOS exhibit the hypertensive pattern: focus on oxidative stress.

Numerous evidence implies complex interrelations between polycystic ovary syndrome (PCOS) and hypertension (HT) in reproductive-age women. In this study, we aimed to investigate the potential strain differences in ovarian morphology, hemodynamic, and biochemical characteristics in an androgen-induced PCOS rat model. A total of 24 rats of 3 weeks old (12 Wistar Kyoto - WK and 12 spontaneously hypertensive rats - SHR) were divided into four groups: WK, WK PCOS, SHR, and SHR PCOS. PCOS was induced by daily s.c. injections of testosterone enanthate (1 mg/100 g body weight) administered for 5 weeks. PCOS induction led to estrus cyclicity cessation, cystic ovarian appearance, and sex hormones disturbances in both strains. The morphometric parameters in ovaries were altered in a manner of PCOS-related changes in both strains (higher number in preantral, atretic, and cystic follicles). Ultrasonographically, a significant decrease in ovarian volume (OV) was registered in PCOS groups but also in SHR compared to WK rats. All blood pressure parameters were higher in SHR compared to WK. PCOS modeling increased systolic, mean arterial, and pulse pressure in WK strain, while in SHR, only mean arterial and pulse pressure were higher. Alterations in oxidative stress parameters could provide a molecular basis for PCOS-related changes: in PCOS groups, thiobarbituric acid reactive substance and superoxide anion radical levels were higher in both strains, while superoxide dismutase and glutathione were significantly lowered.

Effect of chlorogenic acid on follicular development, hormonal status and biomarkers of oxidative stress in rats with polycystic ovary syndrome.

Polycystic ovarian syndrome (PCOS) is a complex endocrine and metabolic disorder. Chlorogenic acid (CGA) bears antioxidant properties with protective effects on different tissues. This study was conducted to evaluate the effect of CGA on follicular development, hormonal status and biomarkers of oxidative stress in a rat model of PCOS. In this experimental study, 18 rats were divided into three equal groups including: control, non-treated PCOS [(estradiol valerate (EV): 40.00 mg kg-1 intramuscularly)], and PCOS-CGA (EV: 40.00 mg kg-1 intramuscularly and CGA: 100 mg kg-1 intraperitoneally once a week for eight consecutive weeks). At the end of treatment period, all rats were anesthetized. Then 5.00 mL blood samples of rats in the three groups were taken and prepared for hormonal analyses and their ovaries were isolated and dissected mechanically free of fat and mesentery. The ovaries underwent the following analyses: Morphological study with Hematoxylin and Eosin staining and biochemical study using the malondialdehyde (MDA) level and total antioxidant activity. Data were analyzed using one-way ANOVA and post hoc Tukey's test. The serum level of luteinizing hormone, estrogen, testosterone, antioxidant capacity, glutathione and the number of cystic follicles in the PCOS group treated with 100 mg kg-1 Chlorogenic acid compared to the non-treated PCOS group were significantly decreased, however, the serum level of follicle stimulating hormone, progesterone, MDA and the number of secondary, graafian follicles and corpus luteum were significantly increased. Chlorogenic acid could be effective in ameliorating follicular development as well as hormonal and biochemical disorders in rats with PCOS.

Investigating the Effect of Crocus sativus (Saffron) Petal Hydro-alcoholic Extract on Ovarian Follicle, Inflammatory Markers, and Antioxidant Enzymes in Mice Model of Polycystic Ovary Syndrome.

Background:Saffron petals have traditionally been used to treat a variety of diseases, such as gynecological diseases, primary dysmenorrhea, and premenstrual syndrome. Polycystic ovary syndrome (PCOS) is a kind of gynecological disease that causes infertility, menopausal and urogenital disorders and saffron petals seem to be an efficient treatment for such disorders.Methods:NMRI mice (total=60, each group n=12) were divided into control, PCOS, and the treatment groups. PCOS and treatment groups were injected with testosterone enanthate (TE=1 mg/kg). After 4 weeks, the treatment group was treated with Saffron Petal Extract (SPE) for 14 days. Ovary and blood samples were collected for histological and serological analyses, and expression of NF-κB, NF-κB p65, and IκB genes was analyzed. Finally, data were analyzed using one-way ANOVA (p<0.05).Results:In this study, the number of corpus luteum decreased in PCOS mice (p<0.001) but increased with SPE treatment (p<0.05, p<0.01, p<0.001). The number of cystic follicles increased in PCOS mice (p<0.001), but decreased with SPE treatment (p<0.05, p<0.001). TNFα, IL1ß, IL6, IL18, and CRP levels increased in PCOS mice (p<0.001), but decreased following SPE treatment (p<0.05, p<0.001). Glutathione (GSH) and glutathione S-transferase (GST) serum levels decreased in PCOS mice (p<0.001), but increased with SPE treatment (p<0.05, p<0.001). The transcriptional level (s) of NF-κB, NF-κB p65, IκB genes changed in PCOS condition (p<0.001), and were regulated by SPE treatment (p<0.05, p<0.01 and p<0.001).Conclusion:The present study shows that SPE improved the PCOS symptoms in mice via increasing antioxidant factors and reducing inflammatory markers in serum.

Cauliflower (Brassicae oleracea) extract restores hormonal imbalance, resumes follicular maturation, and down regulates oxidative stress in oestradiol valerate- induced polycystic ovary syndrome in Sprague-Dawley rats

Background: Polycystic ovary syndrome is a common cause of anovulatory infertility. Objectives: This study investigated the ameliorative effect of ethanolic extract of cauliflower in oestradiol valerate-induced polycystic ovary syndrome in Sprague-Dawley rats. Method: Twenty five female rats weighing between 120-160 g were used. They were grouped into five (Groups 1-5; N=5). Polycystic ovary syndrome was induced using oestradiol valerate (16 mg). Groups1and 2 served as negative and positive control respectively, groups 3-5 received cauliflower (200, 500 and 1025 mg/kg body weight) for four weeks. Animals were sacrificed; blood and ovarian tissues were collected for biochemical analysis and histology. Results: The relative weight of the ovary was increased significantly in all the groups compared to the control. This significantly reduced when high dose cauliflower group was compared to EV-group. MDA and SOD levels increased significantly in EV-only groups compared to control. Treatment with cauliflower successfully reduced MDA and SOD levels in comparison to EV-only groups. On the other hand, GSH and catalase did not show any significant differences when the treatment groups were compared to control. FSH and progesterone reduced significantly while LH, testosterone, and oestrogen levels increased significantly in EV-only groups compared to control. Treatment with cauliflower was able to ameliorate these effects. Histological sections showed cystic follicles in EV- only treated group compared to control. Treatment with cauliflower extract in increasing doses was able to rescue growing follicles from cystic formation. Conclusion: Cauliflower extract restores hormonal imbalance, recues follicles from cystic formation, and combats oxidative stress in oestradiol valerate-induced polycystic ovary syndrome in Sprague-Dawley rats.

Histomorphological Study of Ovarian Lesions with Emphasis on Rare Entities- A Descriptive Study

Introduction: The ovaries are bilateral organs on either side of the uterus. Non neoplastic and neoplastic lesions occur in all age groups and can present with similar clinical and radiological features. Histopathology remains the gold standard for diagnosing and categorising these lesions. Aim: To study the prevalence of ovarian lesions in relation to age and clinical findings. Neoplastic lesions will be histologically classified as per World Health Organisation guidelines. Findings in this study will be compared with that in other studies. Materials and Methods: This is an observational study of all ovarian lesions undertaken in the Department of pathology in NRI Institute of Medical Sciences, Sangivalasa, Vishakhapatnam, Andhra Pradesh, India, from June 2018 to November 2021. Specimens were received as ovarian masses alone, or in combination with hysterectomy with either unilateral or bilateral salpingo- oophorectomy. Relevant clinical information was obtained from the records. The tissues were processed by using an automatic tissue processor, paraffin blocks were made and sections cut were stained with Haematoxylin and Eosin(H&amp;E) and examined under the microscope. The lesions were categorised as non neoplastic and neoplastic. The neoplastic lesions were classified according to the latest 2020 World Health Organisation (WHO) guidelines. Incidence of various lesions was expressed in percentage. Results: A total of 119 specimens were studied. Some of the specimens had bilateral ovaries and the second ovary had a different nonneoplastic lesion in 18 cases. 80 (60.60%) were nonneoplastic lesions and 52 (39.39%) were neoplastic lesions. 5 cases of torsion could not be classified into neoplastic or non-neoplastic due to lack of viable histological features. Non neoplastic lesions were more common in the 31-50 years age group. Benign neoplasms were common in 41-50 years age group. Malignant tumours were more common in the perimenopausal and postmenopausal age group. The most common non neoplastic lesion were 39 cases (48.75%) of follicular cyst followed by 26 cases (32.5%) of corpus luteal cyst, 13 cases (16.25%) of cystic follicles and 2 cases (2.5%) of endometriotic lesions. Of the neoplastic lesions, 47 cases (90.38%) were benign, 2 cases (3.84%) were borderline and 3 cases (5.76%) were malignant tumours. Surface epithelial tumours were 42 cases (80.76%), followed by 6 cases (11.53%) of germ cell tumours and 4 cases (7.69%) of sex cord stromal tumours. Conclusion: Non neoplastic lesions and neoplastic lesions present with similar clinical and radiologic picture and must be differentiated histopathologically. Non neoplastic lesions were more common in the present study. Among the neoplasms, benign tumours were more common and surface epithelial tumours were more common than any other category. Torsion ovary, Seromucinous cystadenoma, atypical endometrioid tumor and granulosa cell tumor were some rare entities that were encountered in the study. Classifying tumours helps in better patient management.

Therapeutic Effects of Eugenol in Polycystic Ovarian Rats Induced by Estradiol Valerate: A Histopathological and A Biochemical Study.

Background:Polycystic ovary syndrome (PCOS) is a common type of endocrinopathy in women which is accompanied by androgens elevation, insulin resistance, and metabolic dysfunction. Eugenol is a phenolic component of clove oil that has an antioxidant, anti-inflammatory, and anti-diabetic activity. The present study aimed to evaluate the therapeutic effects of eugenol on the PCOS models of rats.Materials and Methods:In this experimental study, thirty adults female Wistar rats weighing between 180 and 200 g were used. Estradiol valerate-induced PCOS rats (4 mg/rat) were treated with eugenol (12 and 24 mg/kg) for 28 days. The effects of eugenol were studied on levels of glucose, lipid profile, liver enzymes, reproductive hormones, oxidative stress, and the expression of cyclooxygenase-2 (Cox-2) and peroxisome proliferator-activated receptor alpha (Ppar-α) genes, using biochemical analysis of blood and histopathological evaluation of ovaries.Results:Estradiol valerate-induced PCOS resulted in the formation of cystic follicles in the ovaries, hyperinsulinemia, hyperglycemia, hyperlipidemia, hyperandrogenism, and anovulation. It altered the Cox-2 and Ppar-α gene expression and increased oxidative stress and activities of liver enzymes. Eugenol treatment improved the PCOS-associated endocrine and metabolic disorder and histopathological alterations, mostly through antioxidant, anti-diabetic, anti-hyperlipidemic, and anti-androgenic properties. It showed beneficial effects on serum glucose, serum insulin, fat profile, reproductive hormones, liver activity, oxidative stress, expression of Cox-2 and Ppar-α genes, as well as restoration of normal ovulation in the PCOS animals.Conclusion:Eugenol could represent a promising natural product to prevent PCOS or reduce its symptoms.

Effect of Electroacupuncture on Reproductive Disorders and Insulin Resistance in a Murine Polycystic Ovary Syndrome Model.

Polycystic ovarian syndrome (PCOS) is a common, complex, and heterogeneous endocrine and metabolic disorder. There is no standardized treatment, and it therefore requires individualized therapies according to the symptoms and pathogenesis of each patient. The present study aimed to determine the effect of electroacupuncture at the acupoints Zusanli (ST36), Sanyinjiao (SP6), and Neiguan (PC6) on reproductive disorders and insulin resistance in a murine model of PCOS induced by dehydroepiandrosterone (DHEA). Vaginal smear analysis was used to determine mice estrous cycle; intraperitoneal glucose and insulin tolerance tests were adopted to analyze metabolic characteristics; enzyme-linked immunosorbent assay was used to measure hormone levels; gene expression was quantified with real-time PCR; hematoxylin and eosin staining was used to observe ovarian morphology. We observed disordered estrous cycle, polycystic ovarian morphology, and higher levels of homeostasis model assessment-insulin resistance (HOMA-IR) and testosterone (T), indicating successful modeling of PCOS. DHEA increased levels of estrogen (E2), progesterone (P), testosterone (T), luteinizing hormone (LH), and follicle-stimulating hormone (FSH), and EA treatment restored them to levels seen in the control group. EA reduced the days in estrus caused by DHEA, improved the abnormal sex hormone receptor genes, and attenuated the DHEA-induced histomorphological changes in mouse ovaries. The average expressions of the androgen receptor (AR), estrogen receptor (ER), luteinizing hormone receptor (LHR), and follicle-stimulating hormone receptor (FSHR) genes in the ovary greatly increased after DHEA treatment and significantly decreased in the DHEA + EA group. After EA treatment, the cystic follicle (CF) number was reduced and corpora lutea (CL) increased in the DHEA + EA group compared to the DHEA group. EA improved glucose intolerance and insulin intolerance. Statistical analysis of intraperitoneal glucose tolerance test-area under curve (IPGTT-AUC) glucose levels revealed a significant decrease in DHEA group mice compared to the control and DHEA + EA groups. EA was found to restore fasting blood glucose, fasting serum insulin, and HOMA-IR. In summary, our study suggests that EA has a remarkable effect in the DHEA-induced murine PCOS model. Management of EA could improve estrous cycle, hormonal disorders, abnormal sex hormone receptors in ovaries, ovary morphology, and insulin resistance in PCOS mice.

Bioactive component of aqueous extract of Solanum melongena ameliorate estradiol valerate induced ovarian-pituitary dysfunctions in female Sprague–Dawley rats: Histomorphological and biochemical evidence

BackgroundEstradiol valerate (EV) is known to cause hypothalamic-pituitary dysregulation of the gonadotrophin-releasing hormone. This study focused on the impact of aqueous extract of Solanum melongena(SM) on EV-induced ovarian-pituitary dysfunction in female Sprague-Dawley rats. MethodsThirty-two healthy female Sprague Dawley rats weighing (8 weeks old; weight 150–200 g) were randomly divided into four groups of eight (n = 8) rats each. Group A received 2 ml of distilled water, Group B received a single dose of 4 mg/kg body weight (bwt) of EV (i.p), Group C received 500 mg/kg bwt SM extract orally and Group D received a single dose of 4 mg/kg bwt of EV (i.p) and 500 mg/kg bwt of SM orally, the experiment lasted for 21days. The following parameters body weight, pituitary gland weight, ovary weight, ovary, and pituitary gland histomorphology, total antioxidant status (TOS), total oxidant status (TOS), oxidative stress index (OSI), total cholesterol (TC), triglyceride, low-density lipoprotein (LDL), and high-density lipoprotein (HDL), progesterone, FSH, LH, Testosterone, and estradiol were assessed. ResultsEstradiol valerate decreases the number of healthy follicles, progesterone, estradiol, HDL, and TAS. And increase degenerated cystic follicles, TT, FSH, LH, triglyceride, TC, LAL, TOS, and OSI. Pituitary gland showed degenerative changes, characterized by neutrophil infiltrations and loss of cellular projections marked improvement was observed in the ovary, pituitary gland, hormone, and biochemical parameters after SM administration. ConclusionSolanum melongena offers protection against EV-induced ovarian-pituitary dysfunctions evidence by restoration of oxidative stress markers, steroidogenesis, and histopathological alterations back to control level.

THE THERAPEUTIC EFFECT OF PIPER LONGUM LINN. FRUITS IN TESTOSTERONE INDUCED POLYCYSTIC OVARIAN SYNDROME IN FEMALE WISTAR RATS

The regulation of polycystic ovarian syndrome (PCOS) is intricate but often disappointing. The intend of existing study is to investigate effects hydro alcoholic extract of Piper longum L. (HEPL) fruits in testosterone persuaded Polycystic ovarian syndrome. Female wistar arts were separated into 9 groups (n=6). Normal Control received Sesame oil as a vehicle control once daily. Testosterone (2mg/kg/s.c.) was administered for 31days to induce PCOS induction in all remained group. PCOS persuaded animals were given with Metformin (300mg/kg/oral route), Clomiphene citrate, (150mg/kg/oral), HEPL groups (200, 400 and 800 mg/kg), Combination group (Piper longum 800 mg/kg and Metformin 300mg/kg) and Combination group (Piper longum 800 mg/kg and Clomiphene citrate,150mg/kg) up to 59 days using vehicle as a Distilled water. Body weights, estrus cyclicity, biochemical, hormonal and histological orders were measured. Testosterone persuaded PCOS marked by uneven estrus cyclicity, enhanced triglycerides, cholesterol, LDL glycaemia, testosterone and LH (p&lt; 0.05, 0.001) levels were analyzed. To study insulin resistance, Oral glucose tolerance test was performed. Multiple cysts had been observed with PCOS rats as compared with the control group. Also, HEPL established the estrus periodicity. These biochemical, hormonal and structural alterations were abridged by Piper longum L. Moreover, Piper longum significantly decreased Cystic follicles count, testosterone and LH (p

Molecular regulation of polycystic ovary syndrome: altered gene expression levels in mouse models pretreatment and post-treatment.

Polycystic ovary syndrome (PCOS) is a complex disorder and genetic factors are believed to play a role. The main aim was to investigate expression levels of genes involved in PI3K/AKT signalling pathway pretreatment and post-treatment. Mouse models of PCOS were generated. Group one included control mice with no polycystic ovaries (n = 4), Group 2 included a PCOS mouse model (n = 8), Group 3 included PCOS mice treated with clomiphene citrate (n = 7) and Group 4 included PCOS mice treated with clomiphene citrate, metformin and pioglitazone (n = 8). Histochemical analyses were performed. Total RNA was extracted and cDNA was synthesized. Irs, Akt1 and Akt2, mTor and Pdpk1 gene expression levels were evaluated by RT-PCR amplification. In Group 1, cortex and medulla were evaluated as normal; in Group 2, ovarian cortex was composed of immature oocytes and cystic follicles with atretic follicles. In Groups 3 and 4, follicles were in the process of normal follicle differentiation. The expression levels of Akt1 and Pi3k were significantly different (P < 0.0001) between Groups 1 and 2. The significant differences in expression levels of Pi3k and Akt1 were also observed between the Group 1 and both Groups 3 and 4 (P < 0.0001). Furthermore, significant variations of the expression levels of mTor between Groups 1 and 4 were observed. The extrapolation of results of this study may imply that follicular development may be regulated by molecular pathways involving Pi3k, Akt1 and mTor expression. Therefore, genes in the PI3K/AKT pathway may have a direct regulatory role in the development of PCOS.

Brown Adipose Tissue Activation by Cold Treatment Ameliorates Polycystic Ovary Syndrome in Rat.

Polycystic ovary syndrome (PCOS) is a common endocrine disease accompanied by energetic metabolic imbalance. Because the etiology of PCOS is complex and remains unclear, there is no effective and specific treatment for PCOS. It is often accompanied by various metabolic disorders such as obesity, insulin resistances, and others. Activated brown adipose tissue (BAT) consumes excess energy via thermogenesis, which has positive effects on energy metabolism. Our previous research and that of others indicates that BAT activity is decreased in PCOS patients, and exogenous BAT transplantation can improve PCOS rodents. Notably however, it is difficult to apply this therapeutic strategy in clinical practice. Therapeutic strategies of enhancing endogenous BAT activity and restoring whole-body endocrine homeostasis may be more meaningful for PCOS treatment. In the current study, the dehydroepiandrosterone-induced PCOS rat was exposed to low temperature for 20 days. The results show that cold treatment could reverse acyclicity of the estrous cycle and reduce circulating testosterone and luteinizing hormone in PCOS rats by activating endogenous BAT. It also significantly reduced the expression of steroidogenic enzymes as well as inflammatory factors in the ovaries of PCOS rats. Histological investigations revealed that cold treatment could significantly reduce ovary cystic follicles and increase corpus luteum, indicating that ovulation was recovered to a normal level. Concordant with these results, cold treatment also improved fertility in PCOS rats. Collectively, these findings suggest that cold treatment could be a novel therapeutic strategy for PCOS.

257 Awardee Talk: Pursuit of Novel Factors that Regulate Ovarian Follicular Function

Abstract As follicles grow, theca cells (TC) and granulosa cells (GC) must proliferate with minimal differentiation while thecal vascularization increases so that follicles do not prematurely ovulate or luteinize before the oocyte is fully mature. In the early 2000s we used Affymetrix microarrays to discover several unique genes involved in ovarian follicular development. Thrombospondin and fibroblast growth factor (FGF) 2 receptor genes were stimulated by IGF1 in porcine GC. We compared GC gene expression in bovine cystic versus normal follicles and discovered several novel genes including Indian hedgehog protein (IHH), FGF9, brain ribonuclease (BRB), and G protein-coupled receptor 34 (GPR34), leading to identification of roles for these proteins in ovarian follicular development. During the past 10 years, follow-up TC microarray and mechanistic studies have identified FGF9 control of cell cycle proteins, tight junction proteins, and microRNA 221 (MIR221), and that the mitogenic and steroidogenic responses to the major trophic hormones of the ovary (including IGF1, LH and FSH) are altered by overexpression of MIR221 in GC. In addition, we discovered that: 1) FGF9 stimulates GC and TC mitosis while inhibiting steroidogenesis; 2) FGF9 induces E2 transcription factor (E2F)-1, E2F-8 and cyclin D1 (CCND1), and that both IGF1 and vascular endothelial growth factor-A (VEGFA) synergize with FGF9 to further induce E2F8 and CCND1 mRNA; 3) FGF9 induces the nuclear protein UHRF1; and 4) an E2F inhibitor blocks the stimulatory and inhibitory effects of FGF9 on GC proliferation and steroidogenesis, respectively, and down-regulates UHRF1 mRNA and up-regulates VEGFA mRNA. Thus, aberrant production of FGF9 and the factors it induces/inhibits may lead to vascular dysfunction and ovarian disorders such as ovarian cysts. With additional research, knowledge about these newly identified factors may be used to help the livestock industry improve reproductive efficiency via new treatments for estrous synchronization, superovulation and cystic ovaries.

A novel PCOS rat model and an evaluation of its reproductive, metabolic, and behavioral phenotypes.

BackgroundAlthough animal models of PCOS have been used in many studies, none of them can reproduce both the reproductive and metabolic phenotypes of PCOS. In addition, behavioral parameters have not been evaluated in PCOS animal models.PurposeWe tried to produce an improved rat model of PCOS, and the reproductive, metabolic, and behavioral phenotypes of the model rats were evaluated.MethodsFemale rats were implanted with silicon tubes containing oil‐dissolved dihydrotestosterone (Oil‐DHT) as a new PCOS model. Their phenotypes were compared with those of conventional PCOS model rats (DHT), into which tubes containing crystalline DHT were implanted, and non‐DHT‐treated rats (control).ResultsBoth the Oil‐DHT and DHT rats showed greater body weight gain, food intake, and fat depot weight than the control rats. Furthermore, these groups showed fewer estrous stages and increased numbers of cystic follicles. The DHT rats exhibited lower ovarian and uterine weights than the control rats, whereas no such changes were observed in the Oil‐DHT rats. The Oil‐DHT and DHT rats showed less locomotor activity in the light phase than the control rats.ConclusionsOur proposed PCOS model reproduced both the reproductive and metabolic phenotypes of PCOS and may have potential for PCOS research.

THE POSSIBLE EFFECT OF FLAXSEED EXTRACT ON LETROZOLE-INDUCED POLYCYSTIC OVARY RAT MODEL: CORRELATIVE HISTOLOGICAL AND FUNCTIONAL STUDY

Background: Polycystic Ovary Syndrome (PCOS) is one of the most common metabolic disorders affecting women in the reproductive age leading to infertility/subfertility. It may occur as a complication of Letrozole (LTZ) therapy in breast cancer patients. Objective: This was a preliminary study to assess the possible protective and/or curative effect of flaxseed extract in a letrozole-induced PCO rat model. Material and Methods: Thirty-six adult female albino rats were divided into five main groups: Group I kept as the control group, Group II (Letrozole-induced PCO group), received letrozole solution (3 mg/kg) orally once daily by gastric tube for 21 days. Group III (combined flaxseed and letrozole group), received aqueous flaxseed extract (500 mg /kg/day) orally one hour before letrozole in the same dose and duration as group II, Group IV (recovery group), received letrozole in the same dose and duration as group II and then left without treatment for the next 4 weeks after PCO induction, Group V (flaxseed-treated group), treated with aqueous flaxseed extract orally once daily for the next 4 weeks after PCO induction. At the end of the experiment, final body weight, ovarian weight, blood glucose, serum antioxidants assay, and sex hormones levels were measured. Also, specimens of ovaries were processed for light and electron microscopic studies. Histomorphometric and statistical analyses were done. Results: letrozole-induced PCO resulted in a significant increase in final body weight, ovarian weight as well as abnormal blood glucose, sex steroids hormone levels, and antioxidant assays. Moreover, numerous ovarian cystic follicles, widespread cytoplasmic vacuolation in the granulosa lutein cells, congested blood capillaries, and increased fibrosis within the ovarian medulla were evident in the PCO group in concomitant with a significant increase in area percentage of collagen fibers. The ultrastructural assessment confirmed these distortions. The protective and curative potential of flaxseed is mostly comparable revealed improvement in the final body weight, biochemical markers and histological changes, with the protective effect was more pronounced than the curative one. Conclusion and Recommendation: Flaxseed succeeded in ameliorating letrozole-induced polycystic ovary owing to its phytochemicals, antioxidant and anti-inflammatory properties with folliculogenesis potentials. So, it is recommended to use flaxseed as a co-therapy with letrozole to reduce its hyperandrogenic effects.