Rat Androgen Research Articles

Development and evaluation of novel rodent model of PCOS mimicking clinical phenotype in human disease

BackgroundPolycystic ovary syndrome is a most common female reproductive disorder, involving endocrine and metabolic disorders with unclear etiology. Androgen-based rodent animal models like DHEA and DHT are most suitable for PCOS induction, but still, these models fail to produce non-lean PCOS phenotypes such as hyperandrogenism, hyperinsulinemia, elevated estrogen levels, and ovary weight. Excess fructose consumption leads to hyperandrogenism, hyperinsulinemia, and insulin resistance. The purpose of this study is to investigate, whether fructose consumption along with androgens in rats, could develop all metabolic and endocrine phenotypes of non-lean human PCOS disease. MethodsPrepubertal SD rats were administered with DHT (83ug, s.c.) and fructose (20%, p.o.) for 90 days whereas DHEA (7 mg/kg, s.c) and fructose (20%, p.o.) for 30 days. During study duration, the blood glucose level for oral glucose tolerance test, estrus cyclicity, and ultrasonography was observed. Reproductive hormones LH, FSH, insulin, testosterone, and estradiol levels were assessed using ELISA. The ovary, uterus, abdominal fat, and subcutaneous fat were collected and weighed, and histopathology was done for any anomaly’s findings.ResultsDHT + fructose-treated rats showed significant (p < 0.05) increase in serum testosterone, LH, estradiol, decreased FSH levels, and caused multiple cystic follicles. Abdominal fat, subcutaneous fat, ovary, and uterine weight were higher in DHT + F and DHEA + F when compared to control groups. OGTT reveals impaired insulin sensitivity and glucose tolerance in both model groups. Ovarian histopathology of DHT + F shows more cysts than the DHEA + F groups. No significant changes in uterine histology of DHT + F and DHEA + F-treated rats.ConclusionDHT + F-treated rats mimic all clinical phenotypes and could be used as novel rodent model for non-lean type PCOS.

Rapid and reagent-free bioassay using autobioluminescent yeasts to detect agonistic and antagonistic activities of bisphenols against rat androgen receptor and progesterone receptor

Bisphenol A (BPA) and its analogues have been classified as endocrine disruptors via binding to nuclear receptors. Two novel bioassays, BLYrARS and BLYrPRS, were developed for rapid detection of agonistic and antagonistic activities of BPA and five of its analogues binding rat androgen receptor (rAR) and rat progesterone receptor (rPR). The reporter bioassay was based on two autonomously bioluminescent strains of the yeast Saccharomyces cerevisiae, recombined with a bacterial luciferase reporter gene cassette (lux) that can produce autofluorescence, regulated by the corresponding hormone response element acting as the responsive promoter. The bioluminescent signal is autonomous and continuous without cell lysis or addition of exogenous reagents. The AR agonist R1881 could be detected at 4 h with a half-maximal effective concentration (EC50) of ~9.4 nM. The PR agonist progesterone could be determined at 4 h with an EC50 of ~2.74 nM. None of the sixteen bisphenols presented agonistic activities against rAR and rPR. However, thirteen BPs were rAR antagonists and eleven BPs acted as rPR antagonists with different potency. The BLYrARS and BLYrPRS bioassay characterized by automated signal acquisition without additional manipulations or cost can be applied for simple and rapid detection of agonistic and antagonistic activities of BPs and other compounds acting as agonists or antagonists of rAR and rPR. Based on data derived by use of this bioassay endocrine-disrupting activities of some BPA analogues are more potent than BPA.

Combination of selective androgen and estrogen receptor modulators in orchiectomized rats

PurposeSelective androgen and estrogen receptor modulators, ostarine (OST) and raloxifen (RAL), reportedly improve muscle tissue and offer therapeutic approaches to muscle maintenance in the elderly. The present study evaluated the effects of OST and RAL and their combination on musculoskeletal tissue in orchiectomized rats.MethodsEight-month-old Sprague Dawley rats were analyzed. Experiment I: (1) Untreated non-orchiectomized rats (Non-ORX), (2) untreated orchiectomized rats (ORX), (3) ORX rats treated with OST during weeks 0–18 (OST-P), (4) ORX rats treated with OST during weeks 12–18 (OST-T). Experiment II: 1) Non-ORX, (2) ORX, 3) OST-P, (4) ORX rats treated with RAL, during weeks 0–18 (RAL-P), 5) ORX rats treated with OST + RAL, weeks 0–18 (OST + RAL-P). The average daily doses of OST and RAL were 0.4 and 7 mg/kg body weight (BW). Weight, fiber size, and capillarization of muscles, gene expression, serum markers and the lumbar vertebral body were analyzed.ResultsOST-P exerted favorable effects on muscle weight, expression of myostatin and insulin growth factor-1, but increased prostate weight. OST-T partially improved muscle parameters, showing less effect on the prostate. RAL-P did not show anabolic effects on muscles but improved body constitution by reducing abdominal area, food intake, and BW. OST + RAL-P had an anabolic impact on muscle, reduced androgenic effect on the prostate, and normalized food intake. OST and RAL improved osteoporotic bone.ConclusionsThe OST + RAL treatment appeared to be a promising option in the treatment of androgen-deficient conditions and showed fewer side effects than the respective single treatments.

Dulaglutide, a long-acting GLP-1 receptor agonist, can improve hyperandrogenemia and ovarian function in DHEA-induced PCOS rats

ObjectiveThe purpose of this study was to explore the effect of dulaglutide on DHEA induced PCOS rats and its mechanism, to provide new drugs and research directions for clinical treatment of PCOS. MethodsIn this study, the PCOS model was established by giving female SD rats subcutaneous injection of DHEA for 21 consecutive days. After modeling, the treatment group was injected subcutaneously with three doses of dulaglutide for 3 weeks. The model group was injected with sterile ultrapure water, and the normal group did not get any intervention. The body weight changes of rats in each group were recorded from the first day when rats received the administration of dulaglutide. Three weeks later, the rats were fasted the night after the last treatment, determined fasting insulin and fasting glucose the next day. After the rats were anesthetized by chloral hydrate, more blood was collected from the heart of the rat. The serum insulin, testosterone and sex hormone binding globulin (SHBG) levels were detected by the enzyme-linked immunoassay method. After removing the adipose tissue, the obtained rat ovary tissue was used for subsequent experimental detection, using HE staining for morphology and follicular development analysis; qRT-PCR for the detection of 3βHSD, CYP17α1, CYP19α1, and StAR gene expression in ovarian tissue; and western blotting analysis of CYP17α1, CYP19α1, StAR protein expression and insulin level to verify whether dulaglutide has a therapeutic effect on PCOS in rats. ResultsAfter treated with different concentrations of dulaglutide, we found that the body weight of rats in the treatment groups were reduced. Compared with the rats in PCOS group, the serum androgen level of rats in the treatment groups was significantly decreased, and the serum sex hormone binding protein content was significantly increased, and there was statistically significant difference between these groups and PCOS group. In terms of protein expression and gene regulation, the expression of 3βHSD, CYP19α1 and StAR in the ovarian tissue of rats in treatment groups were decreased significantly after received the treatment of dulaglutide, and there was statistically significant difference between these groups and PCOS group. In addition, dulaglutide reduced the insulin content in the ovarian tissue of PCOS rats. ConclusionDulaglutide may reduce the hyperandrogenemia of PCOS rats by regulating the content of serum SHBG and the expression of 3βHSD, CYP19α1, and StAR related genes and proteins, thereby inhibiting the excessive development of small follicles and the formation of cystic follicles in the ovaries of PCOS rats, thereby improving polycystic ovary in PCOS rats. In addition, dulaglutide may reduce the weight of PCOS rats, further reducing the level of high androgen in PCOS rats, and improving the morphology of their polycystic ovaries.

Neonatal androgenization in rats affects oocyte maturation.

Androgens are relevant in order to achieve a normal growth and maturation of the follicle and oocyte, since both excess and absence of androgens may affect the correct ovarian function. The current study analyzes the impact of neonatal androgenization in the first ovulation and oocyte maturation in response to exogenous gonadotrophin stimulation. Neonatal rats were daily treated with testosterone, dihydrotestosterone, or vehicle during follicle assembly period (days 1 to 5). At juvenile period, rats were stimulated sequentially with PMSG and hCG. Ovulation, ovarian histology, hormonal milieu, morphological characteristics of meiotic spindle, and in vitro fertilization rate in oocytes were analyzed. Our data shows that oocytes from androgenized rats displayed a major proportion of aberrant spindles and altered meiotic advance that control animals. These alterations were accompanied with an increase in both fertilization rate and aberrant embryos after 48 h of culture. Our findings showed a direct impact of neonatal androgens on oocyte development; their effects may be recognized at adulthood, supporting the idea of a programming effect exerted by neonatal androgens. These results could be relevant to explain the low fertility rate seen in polycystic ovary syndrome patients after in vitro fertilization procedures.

Ovariectomy Improves Metabolic and Oxidative Stress Marker Disruption in Androgenized Rats: Possible Approach to Postmenopausal Polycystic Ovary Syndrome.

Background: This study aims to evaluate metabolic and oxidative stress markers in a postmenopausal rat model of polycystic ovary syndrome (PCOS). Methods: Wistar rats were divided in four groups: control ovariectomized (OVX; n = 9), control SHAM (n = 9), androgenized OVX (n = 10), and androgenized SHAM (n = 10). Female rats were androgenized during the neonatal period and compared with controls. Surgery (ovariectomy or SHAM procedure) was performed at day 100 and euthanasia at day 180 of life. Bodyweight, lipids, glucose, triglyceride glucose (TyG) index, and oxidative stress markers (total oxidant status [TOS], total antioxidant capacity, nitric oxide, ferric-reducing ability of plasma [FRAP], and advanced oxidation protein product) were addressed. Results: Androgenized SHAM rats exhibited a higher total, low-density lipoprotein cholesterol, triglycerides, TyG index (an insulin resistance marker), and increased TOS, FRAP, and albumin in comparison with control SHAM rats. These abnormalities disappeared after ovariectomy despite the fact that ovariectomized androgenized rats became heavier than the other three groups. Conclusion: Ovariectomy improved metabolic and oxidative stress markers in a rat model of PCOS.

Acetate causes renoprotection like androgen and mineralocorticoid receptors blockade in testosterone-exposed pregnant rats.

The kidney plays a critical role in human health and deviation from its normal function can lead to severe morbidity and mortality. Exposure to excess testosterone in women has been linked to several disorders, including kidney disorder and acting undoubtedly through androgen receptor (AR), whereas the involvement of mineralocorticoid receptor (MR) is unclear. Likewise, the renal effect of sodium acetate (SAc) during late gestational exposure to testosterone is not well known. We hypothesized that SAc or MR blockade would protect the kidney of testosterone-exposed pregnant rats against glutathione and adenosine depletion. Twenty-five pregnant Wistar rats were treated (sc) with olive oil, testosterone propionate (0.5mg/kg) singly or in combination with SAc (200mg/kg; p.o.), androgen receptor (AR) blocker, flutamide (Flu; 7.5mg/kg; p.o.) or (MR) blocker, eplerenone (Eple; 0.5mg/kg) between gestational days 14 and 19. Glutathione, adenosine and nitric oxide were decreased while uric acid (UA), xanthine oxidase (XO), malondialdehyde (MDA), lactate dehydrogenase activity and free fatty acids were increased in the kidneys of gestational rats exposed to testosterone. Also, plasma urea and creatinine were elevated. SAc and Eple reversed tested testosterone-induced effects in gestational rats. The exposure to testosterone impairs renal antioxidant defense via AR and MR during late gestation in pregnant rats. The study also provides evidence that sodium acetate protects the kidneys of gestational testosterone-exposed rats against defective antioxidant defense in like manner as MR or AR antagonist.

Effects of gonadectomy and dihydrotestosterone on neuronal plasticity in motivation and reward related brain regions in the male rat.

Gonadal hormones affect neuronal morphology to ultimately regulate behaviour. In female rats, oestradiol mediates spine plasticity in hypothalamic and limbic brain structures, contributing to long-lasting effects on motivated behaviour. Parallel effects of androgens in male rats have not been extensively studied. Here, we investigated the effect of both castration and androgen replacement on spine plasticity in the nucleus accumbens shell and core (NAcSh and NAcC), caudate putamen (CPu), medial amygdala (MeA) and medial preoptic nucleus (MPN). Intact and castrated (gonadectomy [GDX]) male rats were treated with dihydrotestosterone (DHT, 1.5mg) or vehicle (oil) in three experimental groups: intact-oil, GDX-oil and GDX-DHT. Spine density and morphology, measured 24hours after injection, were determined through three-dimensional reconstruction of confocal z-stacks of DiI-labelled dendritic segments. We found that GDX decreased spine density in the MPN, which was rescued by DHT treatment. DHT also increased spine density in the MeA in GDX animals compared to intact oil-treated animals. By contrast, DHT decreased spine density in the NAcSh compared to GDX males. No effect on spine density was observed in the NAcC or CPu. Spine length and spine head diameter were unaffected by GDX and DHT in the investigated brain regions. In addition, immunohistochemistry revealed that DHT treatment of GDX animals rapidly increased the number of cell bodies in the NAcSh positive for phosphorylated cAMP response-element binding protein, a downstream messenger of the androgen receptor. These findings indicate that androgen signalling plays a role in the regulation of spine plasticity within neurocircuits involved in motivated behaviours.

Dextran-coated superparamagnetic nanoparticles modified with folate for targeted drug delivery of camptothecin

Drug delivery vehicles based on magnetic nanoparticles present a promising strategy for cancer treatment, due to controlled targeted treatment, high loading efficiency, and biosafety as compared to traditional drug delivery approaches. In the present study, superparamagnetic iron oxide (Fe3O4) nanoparticles (SPIONs) were synthesised by a co-precipitation approach, stabilised with dextran (DEX), and successfully conjugated with folate (FA) for delivery of camptothecin (CPT) in prostate cancer cells. Size and other characteristics of the modified nanoparticles were measured using scanning electron microscopy (SEM), transmission electron microscopy (TEM), dynamic light scattering (DLS), and zeta-potential. The results demonstrated that the prepared FA-DEX-SPIONs were spherical in shape with an average diameter of 63.31 nm. They had anticancer activity with high CPT loading efficiency in AT3B-1cancer cells, enabling the therapeutic activity of the drug via its active delivery and release at 37 °C in phosphate and citrate buffer solutions. MTT results exhibited no cytotoxicity on rat androgen independent prostate cancer (AT3B-1) and normal human prostate (RWPE-1) cells. In conclusion, FA-DEX-SPION composite is a promising candidate that could be used for the targeted release of CPT anti-cancer drug.

Trabecular Bone is Increased in a Rat Model of Polycystic Ovary Syndrome.

Polycystic ovary syndrome (PCOS) in an intricate disorder characterized by reproductive and metabolic abnormalities that may affect bone quality and strength along with the lifespan. The present study analysed the impact of postnatal androgenization (of a single dose of testosterone propionate 1.25 mg subcutaneously at day 5 of life) on bone development and markers of bone metabolism in adult female Wistar rats. Compared with healthy controls, the results of measurements of micro-computed tomography (microCT) of the distal femur of androgenized rats indicated an increased cortical bone volume voxel bone volume to total volume (VOX BV/TV) and higher trabecular number (Tb.n) with reduced trabecular separation (Tb.sp). A large magnitude effect size was observed in the levels of circulating bone formation Procollagen I N-terminal propeptide (P1NP) at day 60 of life; reabsorption cross-linked C-telopeptide of type I collagen (CTX) markers were similar between the androgenized and control rats at days 60 and 110 of life. The analysis of gene expression in bone indicated elements for an increased bone mass such as the reduction of the Dickkopf-1 factor (Dkk1) a negative regulator of osteoblast differentiation (bone formation) and the reduction of Interleukin 1-b (Il1b), an activator of osteoclast differentiation (bone reabsorption). Results from this study highlight the possible role of the developmental programming on bone microarchitecture with reference to young women with PCOS.

SUN-023 Changes in Microct Bone Density and Reduced Bone Formation in a “Postmenopausal” PCOS Rat Model

Polycystic ovary syndrome (PCOS) affects women during their entire lifespan. Evidence from the literature suggests an association of PCOS with decreased bone formation markers (osteocalcin and P1NP), although no conclusive data about the incidence of fractures exist. In the present study, we investigated the consequences of androgenization in rats on bone markers and femur microCT and the changes in these parameters after ovariectomy. This study was approved by the local Animal Ethics Committee. Briefly, Wistar rats (n= 38) were divided in 4 groups: 1) “Control OVX” (single dose of corn oil s.c. at day 5 of life and ovariectomy at day 100, n=9); 2) “Control SHAM” (n=9); 3) “Androgenized OVX”(single dose of testosterone propionate 1.25 mg s.c. at day 5 of life and ovariectomy at day 100, n=10); and 4) “Androgenized SHAM” (n=10). Full characterization of estrous cycles and weight was performed during growth, and all animals were euthanized at day 180 during metestrus/diestrous. Evaluation of glucose levels, lipids, estradiol, P1NP levels (a marker of bone formation), and analysis of the femur micro CT Skyscan 1174 (Aartselaar, Belgium) was performed in at least eight animals of each group. Ovariectomy increased the weight of Androgenized OVX rats on day 180: these animals were heavier than Control OVX, Control SHAM, or Androgenized SHAM (ANOVA p<0.001). However, metabolic changes were observed in ovary-intact Androgenized SHAM rats who exhibited higher total cholesterol (ANOVA p<0.001), increased LDL (ANOVA p=0.03), and elevated TyG index, a marker of insulin resistance (ANOVA p<0.001) against all other three groups. This group (Androgenized SHAM rats)also exhibited an increase in MicroCt bone density (g/cm-3) (mean + SEM) of 1.117 + 0.06 against the other - Control SHAM 0.8433 + 0.03, Control OVX 0.5527 + 0.001, and Androgenized OVX 0.6284 + 0.02 (ANOVA p< 0.001). Although the values of bone density between Control OVX and Androgenized OVX groups were similar, gonadal removal produced a different pattern of bone density reduction between Control OVX and Androgenized OVX (Two-Way ANOVA p=0.001). Moreover, we found P1NP levels significantly decreased in the Androgenized OVX group (mean + SEM) of 58.57 + 4.41 ng/ml against 88.02 + 8.49 ng/ml in Control OVX versus (ANOVA p<0.0001) indicating lower bone formation. Our results suggest that bone and metabolic features of Androgenized rats are affected by ovariectomy with a negative impact on bone formation.

Testosterone and Progesterone Levels, Gene Expression of Androgen and Progesterone Receptors in Albino Male Rats Treated with Phenolic Flower Extract of Hibiscus Rosa - Sinensis L

Hibiscus rosa sinensis L. is one of medicinal plant and belongs to the family Malvaceae , Hibiscus species are used in the treatment of many disease, flowers are used to stimulate hair growth , wounds healing activity, anti-parasitic effect , Antidiabetic anti-hyperlipidemic activity. and have many pharmacological properties including anti-fertility , antipyretic, antispasmodic, antifungal, anti-inflammatory and many more. This study was conducted in the department of Biology /College of Science/University of Babylon/Iraq , to evaluate the Effect of Phenolic Extract of Hibiscus rosa - sinensis L. flowers on some biomarkers and gene expression in Albino male Rats .The overall number of animals used were 32 male rats aged 2-3 months. The results showed a significant decrease (p<0.05) in the level of Testosterone and Progesterone in treated rats as compared with control groups for 30 and 60 days. Also the result revealed a significant decrease ( p< 0.05) in the expression of the androgen receptors and progesterone receptor in the testicular cells treatment group with phenolic compounds of H. rosa- sinensis flowers at a dose of 300mg / kg /day of body weight for 60 days compared to control groups.

Histopathological changes in androgenized ovaries are recovered by melatonin treatment.

Nandrolone decanoate (ND) is a synthetic steroid, which promotes adverse effects on the ovarian tissue, and melatonin (MLT) exhibits a number of beneficial properties in the reproductive system. This study evaluated the general features of the ovarian tissue and the immunoexpression of sex steroid receptors in ND-treated rats that were submitted to short-term melatonin treatment. Adult rats received mineral oil (control group) and ND at doses of 7.5 mg/kg for 15 days (ND-treated group). The treatment with MLT (10mg/kg for 7 days) was given alone, before or in combination with ND. All ND-treated animals showed persistent dioestrus. In the androgenized groups that received MLT, ovarian morphology and size, and the number/area of corpora lutea were recovered. The number of healthy and atretic follicles was recovered when MLT was administered prior to ND; this was similar to the ovaries of control and MLT groups. There was a decrease in estrogen receptors immunostaining in the follicles of androgenized rats that were treated with MLT, and pretreatment with MLT reduced the expression of androgen receptor in atretic follicles and corpora lutea, when compared with ND-treated group. We conclude that MLT treatment recovered the histopathological aspects of the androgenized ovaries, and MLT pretreatment was the most effective.

Metformin improves ovarian follicle dynamics by reducing theca cell proliferation and CYP-17 expression in an androgenized rat model

BackgroundMetformin influences insulin receptor signaling, which might interfere with the proliferation of ovarian follicular structures and steroidogenesis. We hypothesize that reductions in glucose and insulin levels might interfere with CYP-17 expression and histomorphological changes in an androgenized rat model. The aim of this study was to analyze the effect of metformin on CYP-17 expression, follicular dynamics, and proliferative parameters in neonatally androgenized female rats.MethodsThirty-six newborn rats were randomly allocated to the following three groups on the third day of life: control (CG, n = 12), androgenized (GA, n = 12), and androgenized + metformin (GAmet, n = 12). The GA and GAmet animals were administered 0.1 mL of testosterone propionate (1.25 mg/animal) diluted in castor oil (vehicle) in a single dose; the CG rats received a subcutaneous injection of the vehicle in the dorsum. After 90 days, gavage treatment was initiated, distilled water was administered to the CG and GA rats, and metformin (150 mg/kg) was administered to the GAmet animals. The treatment was administered daily for six weeks. Following anesthesia, blood was drawn for biochemical measurements, and the ovaries were removed for histological and immunohistochemical analyses of Ki67, VEGFA and CYP17 expression. The glucose and insulin levels were also measured.ResultsThe comparison of the GA and GAmet animals revealed that metformin decreased the weight as well as the glucose and insulin levels, slowed the proliferation of the theca interna and interstitial cells, as evidenced by Ki-67 and VEGF-A expression, and diminished CYP17 expression in the analyzed ovarian structures. In addition, metformin reduced the number of degenerating follicles and interstitial cells and improved angiogenesis.ConclusionMetformin improves the carbohydrate metabolism, reduces proliferation, and decreases CYP-17 expression in the follicular structures of androgenized rats.

Inhibition of adenosine deaminase and xanthine oxidase activities by sodium acetate ameliorates glucose dysregulation and defective antioxidant defenses induced by gestational excess androgen in rats

Inhibition of adenosine deaminase and xanthine oxidase activities by sodium acetate ameliorates glucose dysregulation and defective antioxidant defenses induced by gestational excess androgen in rats

N‐Butanol and Aqueous Fractions of Red Maca Methanolic Extract Exerts Opposite Effects on Androgen and Oestrogens Receptors (Alpha and Beta) in Rats with Testosterone‐Induced Benign Prostatic Hyperplasia

Benign Prostatic Hyperplasia (BPH) affects, worldwide, 50% of 60-year-old men. The Peruvian plant red maca (Lepidium meyenii) inhibits BPH in rodents. This study aimed to determine the effects of methanolic red maca extract and its n-butanol and aqueous fractions on expression of androgen and oestrogen receptors in rats with testosterone enanthate-induced BPH. Thirty-six rats in six groups were studied. Control group received 2 mL of vehicle orally and 0.1 mL of propylene glycol intramuscularly. The second group received vehicle orally and testosterone enanthate (TE) (25 mg/0.1 mL) intramuscularly in days 1 and 7. The other four groups were BPH-induced with TE and received, during 21 days, 3.78 mg/mL of finasteride, 18.3 mg/mL methanol extract of red maca, 2 mg/mL of n-butanol fraction, or 16.3 mg/mL of aqueous fraction from red maca. Treatments with red maca extract and its n-butanol but not aqueous fraction reduced prostate weight similar to finasteride. All maca treated groups restored the expression of ERβ, but only the aqueous fraction increased androgen receptors and ERα. In conclusion, butanol fraction of red maca reduced prostate size in BPH by restoring expression of ERβ without affecting androgen receptors and ERα. This effect was not observed with aqueous fraction of methanolic extract of red maca.

Dutasteride-mediated morphological changes in the genitourinary tract associated with altered expression patterns of the androgen and estrogen receptors in male rats.

We evaluated the effects of dutasteride on the genitourinary tract using fifteen 8-week-old male Sprague-Dawley rats. Animals were divided into three groups comprising five animals each and treated as follows. Group A was a control group, members of Group B received oral administration of dutasteride 0.1mg/kg/day from the age of 8 to 16weeks, and members of Group C were castrated at the age of 8weeks. All rats were killed at the age of 16weeks for the sample collection of blood, bladder, prostate, seminal vesicles, and penis. Then, we evaluated the pathological examination for evaluating the tissue fibrosis and hormonal receptor expression. The results showed that the mean size of the prostate and seminal vesicles was smaller in Group B and Group C than in Group A. Serum and tissue concentrations of both testosterone and dihydrotestosterone were remarkably reduced in serum and all tissues in Group C compared with Group A. On the other hand, in Group B, only dihydrotestosterone was reduced in serum and penis. Histopathological examination revealed that Group C showed statistically significant histological changes, such as an increase in fibrotic tissue in the bladder, prostate, and penis. Similarly, Group B showed fibrotic changes in the prostate and penis compared with the Group A. Immunofluorescent staining revealed that the androgen receptor was more strongly expressed than the estrogen receptor beta in Group A. On the other hand, in Group C, weak expression of the androgen receptor and strong expression of the estrogen receptor beta was noted. In Group B, these changes were noted in the prostate and penis. These findings suggest that dutasteride cause morphological changes not only in prostate but also in penis. These changes are associated with altered expression patterns of androgen receptor and estrogen receptor.

Ovarian sex steroid receptors and sex hormones in androgenized rats

This study evaluated for the first time the effects of different doses of the anabolic steroid nandrolone decanoate (ND) on the expression of ovarian steroid receptors (AR, ER-α (ESR1) and ER-β (ESR2)) and related sex hormones after treatment and recovery periods in adult rats. The animals were injected subcutaneously with doses of ND (1.87, 3.75, 7.5 or 15 mg/kg b.w.) or mineral oil (control group) for 15 days, and the experimental groups were divided into three periods of evaluation: (a) ND treatment for 15 days, (b) ND treatment and recovery for a period of 30 days and (c) ND treatment and recovery for a period of 60 days. Estrous cycle was monitored daily. At the end of each period, rats were killed for collection of blood and ovaries. Persistent diestrus occurred in all rats during ND treatment and after 30-day recovery. The highest dose of ND was able to maintain all rats arrested at diestrus until 60-day recovery. The expression of steroid receptors varied in a dose- and period-dependent manner, having a more pronounced response with the dose of 15 mg ND/kg. ND treatment increased serum levels of testosterone, 17β-estradiol and dihydrotestosterone, especially at the highest doses of 7.5 and 15 mg ND/kg. No change was observed in the levels of follicle-stimulating hormone (FSH), whereas levels of the luteinizing hormone (LH) varied according to the dose and period. In conclusion, the ovarian sex steroid receptors and sex hormones were restored only at lower doses of ND and after a longer period of recovery.

Early ovarian follicular development in prepubertal Wistar rats acutely exposed to androgens.

Androgens may directly modulate early ovarian follicular development in preantral stages and androgen excess before puberty may disrupt this physiological process. Therefore, the aim of this study was to investigate the dynamics of follicular morphology and circulating androgen and estradiol levels in prepubertal Wistar rats acutely exposed to androgens. Prepubertal female Wistar rats were distributed into three groups: control, equine chorionic gonadotropin (eCG) intervention and eCG plus dehydroepiandrosterone (DHEA) intervention (eCG+DHEA). Serum DHEA, testosterone and estradiol levels were determined, and ovarian morphology and morphometry were assessed. The eCG+DHEA group presented increased serum estradiol and testosterone levels as compared with the control group (P<0.01), and higher serum DHEA concentration v. the eCG-only and control groups (P<0.01). In addition, the eCG+DHEA group had a higher number of, and larger-sized, primary and secondary follicles as compared with the control group (P<0.05). The eCG group presented intermediate values for number and size of primary and secondary follicles, without significant differences as compared with the other two groups. The number of antral follicles was higher in the eCG+DHEA and eCG groups v. controls (P<0.05). The number of primordial, atretic and cystic follicles were similar in all groups. In conclusion, the present experimental model using an acute eCG+DHEA intervention was useful to investigate events involved in initial follicular development under hyperandrogenic conditions, and could provide a reliable tool to study defective follicular development with possible deleterious reproductive consequences later in life.

Orexin Decreases Aromatase Gene Expression in The Hypothalamus of Androgenized Female Rats.

Orexin is a hypothalamic orexigenic neuropeptide, which third cerebral injection of it mainly exerts inhibitory effects on reproductive functions. It increases significantly the Aromatase (Cyp19) gene expression in the hypothalamus of male rats. Aromatase is an enzyme which converts androgens to estradiol in the hypothalamus of rats. Prenatal or neonatal exposure of females to testosterone masculinizes the pattern of Cyp19 mRNA levels in adulthood. In the present study the effects of central injections of orexin-A on hypothalamic Cyp19 gene expression of adult female rats were investigated, while they had been androgenized on third day of postnatal life. In this experimental study, twenty female Wistar rats received subcutaneous injections of testosterone propionate (50 µg/100 µl) on their third day of postnatal life. Adult androgenized rats weighing 180-220 g, received either 3 µl saline or one of 2, 4 or 8 µg/3 µl concentration of orexin via third cerebral ventricle. Five non-androgenized rats, as control group, received intra cerebral ventricle (ICV) injection of 3 µl saline. The hypothalamuses were dissected out and mean Cyp19 mRNA levels were determined by semi-quantitative real time-polymerase chain reaction (PCR) method. Data were analyzed by unpaired t test and one-way ANOVA using SPSS software, version 16. Mean relative Cyp19 mRNA level was significantly increased in the hypothalamus of androgenized compared to non-androgenized female rats. Central injec- tions of 2, 4 or 8 µg/3 µl orexin decreased significantly the hypothalamic Cyp19 mRNA level of androgenized rats compared to androgenized-control groups. The results suggested that the orexin may exert inhibitory effects on the gene expression of Cyp19 in the hypothalamus of neonatal androgenized female rats in adulthood.