Increased Serum Luteinizing Hormone Research Articles

Evaluation of Gymnema sylvestre R. Br. against Letrozole Induced Polycystic Ovarian Syndrome in rats

Objectives: To evaluate the therapeutic potential of Gymnema sylvestre (GS) leaf ethanolic extract against letrozole induced polycystic ovarian syndrome (PCOS) in rats. Methods: Thirty female virgin rats (180 - 250g) with regular estrus cycles were used and divided into five groups of six animals each. Group-I serves as normal control receives 1ml of 0.5% carboxyl methyl cellulose orally; group-II to V were treated with letrozole (1mg/kg/b.w. p.o.) for 21 days to induce PCOS. Induction of PCOS was confirmed by measuring menstrual irregularity. After induction of PCOS, group-II serves as PCOS control, group-III receives metformin (20 mg/kg/b.w. p.o.) as standard, and group- IV to V treated with G. sylvestre ethanolic extract 200mg/kg/b.w. and 400mg/kg/b.w. p.o. respectively for 28 days. After completion of the treatment schedule oral glucose tolerance test was carried out, followed by serum lipid profile (LDL, VLDL, HDL, and TG), and hormonal analysis was carried out. Then the animals have sacrificed and the ovaries were then processed for histological examination. Results: The results of this study exhibited irregularity in the estrus cycle, a significant increase in serum blood glucose, and an altered serum lipid profile in PCOS control. In addition, the PCOS control rats showed a significant increase in serum luteinizing hormone, testosterone, and a decrease in estrogen, follicle-stimulating hormone, and progesterone. These changes were significantly reversed by G. sylvestre in a dose dependent manner. Histopathology of the ovary also showed an almost normal ovary in the treatment groups. Conclusion: In conclusion, the treatment of ethanolic leaf extract of Gymnema sylvestre reduces insulin resistance by improving glucose utilization, regularize menstrual irregularity and promote ovulation by correcting the hormonal irregularities and improve the development of follicles in PCOS condition.

Effects of diet-induced obesity and its correction on sex hormone levels and behavioral featurеs of male Wistar rats

Obesity causes a wide range of pathophysiological consequences in the body, affecting the sphere of hormonal and psycho-emotional regulation. The aim of the study is to investigate the effect of a high-calorie diet on the development of obesity, reproductive status hormone levels and behavioral features of male Wistar rats, as well as the possibility of normalizing the identified shifts with non-medicinal options for obesity correction. Experiments were performed on 80 sexually mature male Wistar rats for 16 weeks and included 6 groups of animals, taking into account the diet (standard vivarium diet, highcalorie diet and transition from a high-calorie diet to a standard diet at the 2nd stage of the experiment) with or without the moderate physical activity in the form of treadmill running at the 2nd stage – the last 8 weeks of the experiment. A longterm high-caloric diet (16 weeks) caused visceral obesity, elevated glucose, cholesterol and triglyceride levels, dramatically decreased serum testosterone and estradiol levels, and depression manifestations in the Porsolt test in male Wistar rats. Successful correction of obesity, metabolic shifts and depressive behavior in rats was achieved by switching to a balanced diet during the last 8 weeks of the experiment. Normalization of testosterone and estradiol levels against the background of an increased serum luteinizing hormone level occurred in rats receiving a high-calorie diet (16 weeks) with the connection of physical exercise during the last 8 weeks of the experiment. Complex correction of obesity, which included the transition to a standard diet and exercise during the last 8 weeks of the experiment, led to a dramatic 2-fold increase in testosterone levels compared to control and a hyperactive phenotype in male Wistar rats. There is a close relationship between the depression development and testosterone levels in obesity and its correction.

Beneficial effects of irisin in experimental paroxetine-induced hyperprolactinemia.

It is known that selective serotonin reuptake inhibitors (SSRIs), represent an important and effective treatment of depression and other psychological disorders, these medications can increase prolactin levels mainly through activation of the serotonergic pathway. In this study, we aimed to determine the beneficial effects of irisin on paroxetine, a SSRI, induced hyperprolectinemia and in some other reproductive hormonal changes associated with hyperprolactinemia. Thirty two male Spraque-Dawley rats were used and divided into four groups including sham-operated control (vehicle), irisin (100 ng/kg/day for 28 days with mini-osmotic pumps), paroxetine (treated with 20 mg/kg paroxetine by oral gavage), irisin and paroxetine+irisin groups (n = 8). Serum prolactin (PRL), kisspeptin, luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone and 5-alpha reductase levels were determined with enzyme-linked immunosorbent analysis (ELISA). In animals treated with paroxetine, PRL level increased and testosterone level decreased significantly (p < 0.05). Serum LH level was significantly increased in the group, but no significant changes were observed in the FSH, kisspeptin and 5-alpha reductase levels. Serum prolactin levels was significantly decreased in the group treated with irisin. While no significant difference was observed in kisspeptin, FSH and 5-alpha reductase levels, an increase in serum LH and testosterone levels with irisin administration (p < 0.05). In conclusion, chronic irisin exposure may reverse paroxetine-induced hyperprolactinemia. These results indicate that irisin may have the potential to be used as a therapeutic agent by primarily affecting paroxetine-induced increased prolactin and decreased testosterone levels.

Combined effects of Gymnema sylvestre and Pergularia daemia on letrozole-induced polycystic ovarian syndrome in rats

Objective: To investigate the combined therapeutic potential of Gymnema (G.) sylvestre and Pergularia (P.) daemia on letrozole-induced polycystic ovarian syndrome (PCOS) in rats. Methods: Thirty six healthy female Wistar rats with regular estrus cycles were randomly divided into six groups each of 6. Group I received 1 mL of 0.5% carboxyl methyl cellulose orally and served as the vehicle control group, while groups II to VI were treated with letrozole (1 mg/kg body weight p. o.) for 21 days to induce PCOS. After induction of PCOS, group II served as the PCOS control group, without treatment; group III received metformin (20 mg/kg body weight p. o.) as the standard group, and groups IV to VI received G. sylvestre (100 mg/kg body weight p. o.), P. daemia (300 mg/kg body weight p. o.), and the combination of G. sylvestre and P. daemia, respectively, for 28 days. Vaginal smears were collected from all rats daily throughout the study to determine the phases of the estrus cycle. After completing the treatment schedule, oral glucose tolerance test, serum lipid profile and reproductive hormonal analysis were carried out. Subsequently, the rats were sacrificed to collect ovary and uterus for histopathological examination. Results: The PCOS control rats showed a significant irregularity in the estrus cycle, hyperglycemia, and the altered serum lipid profile such as the increased low and very low density lipoprotein, triglyceride, and decreased high density lipoproteins. In addition, the PCOS control rats showed a significant increase in serum luteinizing hormone, testosterone, and estrogen, and decrease in follicle stimulating hormone and progesterone. These changes were significantly revoked in all the treatment groups. The test drugs also significantly reduced the gained ovary weight (P&lt;0.001), and histopathology of the ovary showed almost normal ovary. Among the treatment groups, the group of combination treatment of G. sylvestre and P. daemia showed superior ameliorative results in PCOS parameters. Conclusions: Combination of G. sylvestre and P. daemia presents potent synergistic activity against hyperandrogenism, hyperinsulinemia, anovulation and follicular cysts in letrozole-induced PCOS rats.

Kigelia Africana Stem Bark Extract Treatment Reverses Type 1 Model of Experimental Diabetes and Associated Reproductive Impairments in Male Wistar Rats

A key hallmark in diabetes is altered reproductive functions. Kigelia africana is reported to offer therapeutic influence in diabetes. However, literature offers nothing concerning its ability to reverse reproductive complications of diabetes. This study investigates the effect of the ethanolic extract of Kigelia africana (EEKA) on reproductive functions in streptozotocin-induced diabetic male rats. Thirty adult male Wistar rats (150-200 g) were randomly divided into five groups; Group 1 (control) received distilled water (vehicle). Group 2 was treated with 250 mg/kg EEKA. Groups 3, 4, and 5 were made diabetic using streptozotocin. Thereafter, groups 4 and 5 were treated with 250 mg/kg EEK and glibenclamide respectively. Treatments were by oral gavage for six weeks. Bodyweight and blood glucose, liver enzymes and proteins, superoxide dismutase (SOD), malondialdehyde activity (MDA), testosterone, follicle-stimulating hormone (FSH), luteinizing hormone (LH), sperm count, and motility was assessed. EEKA treatment significantly reduced blood glucose in diabetic rats. Serum testosterone was significantly reduced in the diabetic group. However, treatment with EEKA significantly modulated the reduction. EEKA treatment in diabetic rats significantly increased serum LH. Therapeutic EEKA treatment in diabetic rats significantly reversed reduced sperm count and motility. EEKA treatment in the diabetic group reduced MDA activity while significantly enhancing SOD activity. The significant increases in serum alanine aminotransferase and aspartate aminotransferase were significantly reduced with EEKA treatment in the diabetic group. Total protein and albumin were significantly reduced in the diabetic group but significantly reversed with EEKA treatment. Therapeutic administration of EEKA reduced hyperglycemia, enhanced sperm count, motility, and serum testosterone levels in diabetic rats. Phytochemical constituents and robust antioxidant activity of EEKA is involved in the modulation and reversal of diabetes and impaired reproductive parameters.

Hormonal profile and characteristics of polychistic ovarium syndrome among Madurese tribe, Indonesia

Polycystic ovarium syndrome (PCOS), the most common endocrine disorders in women of reproductive age, characterized by menstrual disorders (amenorrhea/oligomenorrhea), hirsutism, the appearance of acne, alopecia and the results of biochemical tests that show increased androgens (testosterone). Increased serum luteinizing hormone (LH) and follicle stimulating hormone (FSH) serve as diagnostic tests for PCOS over the years, but from several research results obtained inconsistent results that need further research. The purpose of this study is to analyze the hormonal profile and characteristics of polycystic ovary syndrome (PCOS) in Madura. Case-control study conducted in March-August 2019 in the Madura tribe. There were 32 subjects with PCOS and 32 healthy women participated in this study. Hormonal examination was using a serum and followed by enzyme-linked immune sorbent assay (ELISA kit). Levels Follicle-stimulating hormone (FSH) average was 8.74 and SHBGs were 10.02 lower in PCOS patients and LH levels were higher in PCOS patients; ratio LH/FSH was 0.76. The results of the Madurese study showed that levels of FSH, LH, weight significantly related to PCOS sex hormone binding globulin (SHBG), however body mass index (BMI) levels were not related to PCOS.

Exposure to di-n-octyl phthalate during puberty induces hypergonadotropic hypogonadism caused by Leydig cell hyperplasia but reduced steroidogenic function in male rats

Humans are exposed to phthalates ubiquitously, which may threaten health. However, whether di-n-octyl phthalate can prevent pubertal sexual maturity is still elusive. In this study, male Sprague Dawley rats (age 35 days) were treated daily by gavage with 0, 10, 100, and 1000 mg/kg body weight of di-n-octyl phthalate from day 35 to day 49 after birth. Di-n-octyl phthalate significantly reduced serum testosterone levels at doses of 100 and 1000 mg/kg, but increased serum luteinizing hormone levels of 1000 mg/kg and decreased testosterone/luteinizing hormone ratio at ≥10 mg/kg, without affecting serum follicle-stimulating hormone levels. Di-n-octyl phthalate significantly induced Leydig cell hyperplasia (increased number of CYP11A1-positive Leydig cells) at 100 and 1000 mg/kg. Di-n-octyl phthalate down-regulates the gene expression of Cyp11a1, Hsd3b1 and Insl3 in individual Leydig cells. Di-n-octyl phthalate can also reduce the number of sperm in the epididymis. Di-n-octyl phthalate increased phosphorylated AKT1/AKT2 without affecting their total proteins, but increased the total protein and phosphorylated protein of ERK1/2 and GSK-3β. Primary immature Leydig cells isolated from 35-day-old rats were treated with 0–50 μM di-n-octyl phthalate for 3 h. This phthalate inhibited androgen production under basal, LH-stimulated, and cAMP-stimulated conditions by 5 and 50 μM in vitro via down-regulating Cyp11a1 expression but up-regulating Srd5a1 expression in vitro. In conclusion, di-n-octyl phthalate induces hypergonadotropic hypogonadism caused by Leydig cell hyperplasia but reduced steroidogenic function and prevents sperm production.

Longitudinal Increases in Serum Insulin-like Factor 3 and Testosterone Determined by LC-MS/MS in Pubertal Danish Boys.

Serum concentrations of the peptide hormone insulin-like factor 3 (INSL3) is a candidate marker for improved distinction between constitutional delay of growth and puberty (CDGP) and permanent hypogonadotropic hypogonadism (HH) in boys. To assess the possible diagnostic role of LC-MS/MS-based INSL3 measurements as a marker of imminent puberty by comparison with testosterone (T) and luteinizing hormone (LH) levels in serum longitudinally collected from 18 healthy boys throughout puberty. The first increase in serum LH was detected on average 4 months earlier, as compared with the first observed increases in INSL3 and T. When comparing the 2 testicular hormones only, we found that in 22% (4 of 18) of the boys the first increase in serum INSL3 was observed prior to the first observed increase in T, whereas in 44% (8 of 18) the first increase in T was observed before the first observed increase in INSL3. In the remaining 6 boys, the 2 testicular hormones showed the first increase at the same examination. In some boys with delayed puberty, the first indication of testicular maturation may be detectable by observing serum INSL3. Further studies of LC-MS/MS determination of serum INSL3 in patients with CDGP and HH are warranted.

Irisin may have a role in pubertal development and regulation of reproductive function in rats.

Physical exercise and body muscle/fat mass are known to affect the endocrine system, puberty onset and reproductive health. However, the potential effects of irisin, an adipo-myokine and exercise-induced hormone, have not yet been fully elucidated on reproductive maturation. Therefore, the present study aimed to determine the effects of irisin administration on pubertal maturation and reproductive system in female and male rats. Daily i.p. injection of irisin (100 ng/kg; from postnatal day 21 for about 10 weeks) delayed the ages at the vaginal opening (as an external index of puberty onset) and first estrus. Furthermore, continuous administration of irisin to female rats caused a significant decrease in serum follicle-stimulating hormone levels and an increase in serum luteinizing hormone and 17β-estradiol levels, as well as causing histopathological changes in the ovarian tissue. On the contrary, irisin administration to male rats did not modify the timing of puberty, as estimated by age at preputial separation. However, chronic exposure to irisin produced significant increases in serum luteinizing hormone and testosterone levels and also sperm concentration and seminiferous tubule diameter in male rats. In conclusion, irisin exposure has different effects on both pubertal maturation and reproductive system in female and male rats. The present findings reveal that chronic irisin exposure may lead to disorders in the female reproductive system and may have androgenic potential on the hypothalamic-pituitary-gonadal axis in males.

Erythropoietin-producing hepatocellular receptor A7 restrains estrogen negative feedback of luteinizing hormone via ephrin A5 in the hypothalamus of female rats.

We have previously shown that systemic injection of erythropoietin-producing hepatocellular receptor A7 (EPHA7)-Fc raises serum luteinizing hormone (LH) levels before ovulation in female rats, indicating the induction of EPHA7 in ovulation. In this study, we aimed to identify the mechanism and hypothalamus-pituitary-ovary (HPO) axis level underlying the promotion of LH secretion by EPHA7. Using an ovariectomized (OVX) rat model, in conjunction with low-dose 17β-estradiol (E2) treatment, we investigated the association between EPHA7-ephrin (EFN)A5 signaling and E2 negative feedback. Various rat models (OVX, E2-treated OVX, and abarelix treated) were injected with the recombinant EPHA7-Fc protein through the caudal vein to investigate the molecular mechanism underlying the promotion of LH secretion by EPHA7. Efna5 was observed strongly expressed in the arcuate nucleus of the female rat by using RNAscope in situ hybridization. Our results indicated that E2, combined with estrogen receptor (ER)α, but not ERβ, inhibited Efna5 and gonadotropin-releasing hormone 1 (Gnrh1) expressions in the hypothalamus. In addition, the systemic administration of EPHA7-Fc restrained the inhibition of Efna5 and Gnrh1 by E2, resulting in increased Efna5 and Gnrh1 expressions in the hypothalamus as well as increased serum LH levels. Collectively, our findings demonstrated the involvement of EPHA7-EFNA5 signaling in the regulation of LH and the E2 negative feedback pathway in the hypothalamus, highlighting the functional role of EPHA7 in female reproduction.

Correcting imbalance of sex hormones by a phosphodiesterase 5 inhibitor improves copulatory dysfunction in male rats with type 2 diabetes

IntroductionSexual dysfunction is a common complication in men with type 2 diabetes and is often refractory to treatment. This study investigated the long-term influence of the phosphodiesterase 5 inhibitor (PDE5I)...

Intravenous infusion of kisspeptin increased serum luteinizing hormone acutely and decreased serum follicle stimulating hormone chronically in prepubertal bull calves

Kisspeptin (KP) is a hypothalamic neuropeptide that stimulates the secretion of gonadotropin releasing hormone. To determine the acute and chronic effects of KP on serum concentrations of luteinizing hormone (LH) and follicle stimulating hormone (FSH), prepubertal bull calves [12 ± 1 (SD) weeks of age; 96.5 ± 14.5 kg BW] were administered one of four treatments [0.0 (control; CON), 0.125 (L-KP), 0.25 (M-KP), or 0.5 (H-KP) μg of KP/kg BW/hour] by intravenous infusion for 76 h. Blood samples were collected every 15 min for the first (acute; 1–6 h; Day 1) and last (chronic; 71–76 h; Day 4) 6 h of the intravenous infusions. Serum concentrations of LH and FSH were determined by radioimmunoassay. For each day, effects of treatment, time, and interactions on LH and FSH concentrations and pulse parameters were analyzed using procedures for repeated measures with JMP Software (SAS Inst. Inc., Cary, NC). There was a treatment effect (P = 0.002) and a treatment × time interaction during Day 1 (P = 0.02) such that LH concentrations were greatest following administration of all doses of KP when compared to CON. However, there was no treatment effect (P = 0.57) or a treatment × time interaction during Day 4 (P = 0.20) on serum LH concentrations. There was a treatment by day interaction (P = 0.02) on mean serum FSH concentrations. Most notably, on Day 4 mean serum FSH concentrations during intravenous infusion of M-KP and H-KP doses were less than that of CON. There was a treatment by day interaction (P = 0.0054) on FSH pulse amplitude concentrations, such that intravenous infusion of all doses of KP on Day 4 decreased FSH pulse amplitudes. In conclusion, acute infusion of KP increased LH concentrations and chronic infusion of KP decreased FSH concentrations. Despite the potential suppression of the hypothalamic–pituitary–gonadal axis with chronic infusion of KP, there are likely applications of KP, KP analogs, or KP receptor agonists to hasten the onset of puberty in livestock.

Primary aldosteronism in Klinefelter's syndrome: two cases.

SummaryPrimary aldosteronism (PA) is more common than expected. Aberrant adrenal expression of luteinizing hormone (LH) receptor in patients with PA has been reported; however, its physiological role on the development of PA is still unknown. Herein, we report two unique cases of PA in patients with untreated Klinefelter’s syndrome, characterized as increased serum LH, suggesting a possible contribution of the syndrome to PA development. Case 1 was a 39-year-old man with obesity and hypertension since his 20s. His plasma aldosterone concentration (PAC) and renin activity (PRA) were 220 pg/mL and 0.4 ng/mL/h, respectively. He was diagnosed as having bilateral PA by confirmatory tests and adrenal venous sampling (AVS). Klinefelter’s syndrome was suspected as he showed gynecomastia and small testes, and it was confirmed on the basis of a low serum total testosterone level (57.3 ng/dL), high serum LH level (50.9 mIU/mL), and chromosome analysis. Case 2 was a 28-year-old man who had untreated Klinefelter’s syndrome diagnosed in his childhood and a 2-year history of hypertension and hypokalemia. PAC and PRA were 247 pg/mL and 0.3 ng/mL/h, respectively. He was diagnosed as having a 10 mm-sized aldosterone-producing adenoma (APA) by AVS. In the APA, immunohistochemical analysis showed co-expression of LH receptor and CYP11B2. Our cases of untreated Klinefelter’s syndrome complicated with PA suggest that increased serum LH levels and adipose tissues, caused by primary hypogonadism, could contribute to PA development. The possible complication of PA in hypertensive patients with Klinefelter’s syndrome should be carefully considered.Learning points:The pathogenesis of primary aldosteronism is still unclear.Expression of luteinizing hormone receptor has been reported in aldosterone-producing adenoma.Serum luteinizing hormone, which is increased in patients with Klinefelter’s syndrome, might contribute to the development of primary aldosteronism.

Effects of letrozole administration on growth and reproductive performance in Markhoz goat bucklings

This study evaluated the growth performance, testicular and semen characteristics, and hormonal profile of Markhoz (Iranian Angora) bucklings injected with letrozole (LTZ). Twenty-eight 4–4.5 month old bucks were randomly assigned into four groups and received either 0.25 mg/kg body weight (BW) LTZ subcutaneously (sc LTZ) or intramuscularly (im LTZ), and also sc (sc CONT) or im (im CONT) controls every week for 3 months. The study was performed at the beginning of the breeding season in Sanandaj Animal Husbandry Research Station (46.99 °E, 35.31 °N). The results showed that LTZ causes increased final body weight (25.78 ± 1.61 kg), higher average daily gain (104 ± 0.03 g/days), and decreased feed conversion ratio (7.81 ± 2.57) (P < 0.05). The pre-slaughter, hot, and cold carcass weights (27.56 ± 2.40, 11.45 ± 1.07 and 11.11 ± 1.05 kg, respectively) were (P < 0.05) heavier in LTZ groups while other carcass characteristics did not differ between groups. No differences occurred between the groups in biochemical parameters, except high-density lipoprotein levels (35.47 ± 2.43 mg/dL) which was higher in LTZ treatments (P < 0.05). LTZ-treated bucks had larger scrotal circumference (20.12 ± 5.75 cm), higher relative testicular weight (560.91 ± 78.59 mg/100 g BW) and volume (175.5 ± 29.71 cm3), greater diameter of seminiferous tubules (224.5 ± 5.21 μm), and number of Sertoli cells (8.39 ± 0.77) (P < 0.05). Semen volume (0.74 ± 0.16 mL), sperm concentration (2.64 ± 0.19 × 10−9/mL), total sperm per ejaculate (1.95 ± 0.49 × 10−9), and semen index (1248 ± 323) increased (P < 0.05) by LTZ treatments, while semen pH (6.77), motility (80.91%), progressive motility (76.75%), viability (83.35%), abnormality (13.70%), acrosome integrity (78.06%), and membrane integrity (80.05%) of sperm remained unaffected. Intratesticular and serum testosterone (T) levels (7.97 ± 0.89 ng/mg protein and 2.47 ± 0.59 ng/mL, respectively), serum luteinizing hormone (LH), growth hormone (GH) levels (1.71 ± 0.24 and 3.62 ± 0.33 ng/mL, respectively) of LTZ groups were elevated, whereas intratesticular and serum estradiol (E2) levels (84.14 ± 8.15 pg/mg protein and 32.33 ± 2.16 pg/mL, respectively) decreased (P < 0.05). No differences were recorded between the sc and im routes of LTZ administration in the measured parameters. To conclude, we have found that LTZ treatment improves growth and reproductive functions of goat bucklings associated with increased serum LH and GH, elevated T and reduced E2 levels in both serum and testis.

Dimethoate blocks pubertal differentiation of Leydig cells in rats.

Dimethoate is an organophosphate pesticide. It is widely used in agriculture. However, whether it blocks pubertal development of Leydig cells remains unknown. In the current study, we exposed male Sprague Dawley rats with 7.5 and 15 mg kg-1 dimethoate from postnatal day 35-56. We also exposed Leydig cells isolated from 35-day-old rats for 3 h. Dimethoate reduced serum testosterone levels at 7.5 and 15 mg kg-1 but increased serum luteinizing hormone and follicle stimulating hormone levels at 15 mg kg-1. Dimethoate did not influence Leydig cell number but reduced Leydig cell size and down-regulated Star, Cyp11a1, and Hsd3b1 in Leydig cells as well as their protein expression. Dimethoate inhibited basal androgen output in a dose-dependent manner with the inhibition starting at 0.05 μM. It significantly inhibited luteinizing hormone and 8Br-cAMP stimulated androgen outputs at 50 μM. It significantly inhibited 22R-hydroxycholesterol and progesterone-mediated androgen outputs at 50 μM. Further study demonstrated that dimethoate also down-regulated the expression of Star, Cyp11a1, and Hsd3b1 at 5 or 50 μM invitro. Dimethoate did not directly inhibit rat testicular steroidogenic enzyme activities at 50 μM. In conclusion, dimethoate targets Star, Cyp11a1, and Hsd3b1 transcription, thus blocking Leydig cell differentiation during puberty.

The androgen receptor in the hypothalamus positively regulates hind-limb muscle mass and voluntary physical activity in adult male mice

We have previously shown that expression of the androgen receptor (AR) in neurons within the brain positively regulates hind-limb muscle mass and physical activity in male mice. To further investigate the region of the brain responsible for mediating these effects of testosterone and to determine whether they are only important for muscle mass accrual during development or whether they are also important for the maintenance of muscle mass in the adult, we deleted the AR specifically in the hypothalamus of adult male mice (Hyp-ARKOs). Hyp-ARKO mice were generated by bilateral stereotaxic microinjection of an adeno-associated virus (AAV) expressing GFP and iCre recombinase under the control of the e-synapsin promoter into the hypothalamus of 10-week-old exon 3-AR floxed male mice. AR mRNA was deleted by 45% in the hypothalamus of Hyp-ARKOs at 5 weeks post-AAV-eSyn-iCre injection. This led to an increase in the mass of the androgen-dependent organs, seminal vesicles and kidneys, by 30% (P < 0.01) and 10% (P < 0.05) respectively, and an increase in serum luteinizing hormone (LH) by 2 fold (P < 0.05). Whilst the mean value for serum testosterone was higher in the Hyp-ARKOs, this did not reach statistical significance. Despite a phenotype consistent with increased androgen bioactivity in Hyp-ARKOs, which would be expected to increase muscle mass, the mass of the hind-limb muscles, gastrocnemius (Gast) (P = 0.001), extensor digitorum longus (EDL) (P < 0.001) and soleus (Sol) (P < 0.01) were paradoxically decreased by 12–19% compared to controls. Voluntary physical activity was reduced by 65% (P < 0.05) in Hyp-ARKO male mice and was associated with a reduction in gene expression of Drd1a and Maob (P ≤ 0.05) in the hypothalamus, suggesting involvement of the brain dopaminergic system. These data provide compelling evidence that androgen signalling via the AR in the hypothalamus acts to positively regulate the maintenance of hind-limb muscle mass and voluntary activity in adult male mice, independent of AR signalling in peripheral tissues.

Oxidative Stress and Reproductive Damage Induced by Lead Acetate in Female Guinea Pig (Cavia porcellus): Curative Effects of Hydroethanolic Extract of Spirulina platensis

This study was aimed at evaluating the curative effects of Hydroethanolic Extract Of Spirulina platensis (HESP) on the reproductive function of female Guinea pig exposed to oxidative stress. Sixty females, 3-4 months old, weighing 300-400 g were divided into six groups (10 animals/group). The neutral control received distilled water, the negative control was treated with lead acetate at a dose 12 mg/kg.b.w while the positive control was given 12 mg of lead acetate/kg b.w and 100 mg of vitamin C. Groups 4, 5 and 6 were treated with lead acetate at a dose of 12 mg/kg b.w for the first 30 days and then received from the 31st day to the 90th day HESP at doses of 50,100 and 200 mg/kg.b.w respectively once daily. Results revealed that lead caused prominent toxic effects on fertility, deterioration of sex organs as well as a disruption of serum levels of Luteinizing Hormone (LH), Follicle Stimulating Hormone (FSH) and progesterone. Lead acetate markedly increased tissues oxidative-stress marker (malondiadehyde), whereas it reduces the activities of antioxidant-enzymes, Superoxide Dismutase (SOD), total peroxidase and catalase. These changes were also accompanied by a significant (p<0.05) decrease of body weight gain. However treatment with HESP ameliorated lead acetate-induced anomalies by significantly (p<0.05) increasing the body weight gain, organ weights and feed intake. Also HESP let to a significant (p<0.05) improvement of fertility indices, a significant (p<0.05) increase in serum LH, FSH, progesterone hormones and an increase in antioxidant enzymes. Curative treatment brought about histological tissues protection and a significant (p<0.05) decrease of toxicity biomarkers (AST, ALT, creatinine and urea) and Malondiadehyde (MDA). In conclusion, lead acetate induced reproductive stress and administration of HESP can mitigate these adverse effects due to its antioxidant properties.

Dibutyltin Dichloride Retards Leydig Cell Developmental Regeneration in Adult Rat Testis.

Dibutyltin dichloride (DBTCl), widely used as plastic stabilizer, can cause comprehensive toxicity. The present study aims to investigate the effects of DBTCl on rat Leydig cell developmental regeneration and characterize the related mechanism. Adult male Sprague Dawley rats were randomly divided into four groups and gavaged with saline (control) or 5, 10, or 20 mg/kg/day of DBTCl consecutively for 10 days. At the end of the DBTCl treatment, all rats received a single intraperitoneal injection (i.p.,) of 75 mg/kg ethane dimethane sulfonate (EDS) to eliminate all the adult Leydig cells and to induce Leydig cell developmental regeneration. Leydig cell developmental regeneration was evaluated by measuring the levels of serum testosterone, luteinizing hormone, and follicle-stimulating hormone on days 7, 35, and 56 post-EDS. Leydig cell gene and protein expression levels, as well as cell morphology and cell counts were also carried out on day 56 post-EDS. The present study found that DBTCl significantly reduced serum testosterone levels on days 35 and 56 post-EDS, but increased serum luteinizing hormone (LH) and follicle-stimulating hormone (FSH) levels on day 56 at ≥ 5 mg/kg/day. The mRNA and protein levels of Leydig (Lhcgr, Scarb1, Star, Cyp11a1, Hsd17b3, and Hsd11b1) and Sertoli cells (Fshr, Amh, and Sox9) were significantly downregulated in the DBTCl-treated testes compared to the control. Immunohistochemical staining showed that DBTCl-treatment caused fewer regenerated Leydig cells and impaired Sertoli cell development and function in the testis on day 56 post-EDS. In conclusion, the present study demonstrates that DBTCl retards rat Leydig cell developmental regeneration by downregulating steroidogenesis-related enzymes at the gene and protein levels, inhibiting Leydig cell proliferation and impairing Sertoli cell function and development.

Effect of Acupuncture on Reproductive Endocrine and p-Akt Protein Expression in Polycystic Ovary Syndrome Rats with Insulin Resistance

To observe the effect of manual acupuncture intervention on the reproductive endocrine and p-Akt protein expression in polycystic ovary syndrome with insulin resistance (PCOS-IR) rats, so as to reveal its mechanism in relieving PCOS-IR. Female Wistar rats were randomly divided into control, model, medication (Metformin) and acupuncture groups (n＝8 in each). The PCOS-IR model was established by subcutaneous injection of Dehydroepiandrosterone (DHEA, 6 mg/100 g), once daily for 20 days. Rats of the medication group were treated by gavage of Metformin solution (100 mg/kg), once daily for 28 days. For rats of the acupuncture group, manual acupuncture stimulation (lifting-thrusting and rotating of needles) was applied to the bilateral "Shenshu"(BL 23), "Yishu"(EX-B 3), "Zigong" (EX-CA 1), "Sanyinjiao"(SP 6), "Fenglong"(ST 40) and "Guanyuan"(CV 4) for 5 min, once a day for 28 days. H．E. staining was used to observe the histopathologic changes of the ovary tissue. Serum levels of follicle-stimulating hormone (FSH), estradiol (E2), luteinizing hormone (LH), testosterone (T) and fasting insulin (FINS) were measured by ELISA. Fasting plasma glucose (FPG) was measured by blood glucose meter, and homeostasis model assessment-insulin resistance index (HOMA-IR) was calculated. Western blot was used to detect the expression of Akt and p-Akt proteins in ovarian tissues. Following modeling, the ovaries showed polycystic changes and an increase of atresia follicles with a larger diameter and no corpus luteum in the model group. After the intervention, there were various types of follicles and a small amount of corpus luteum at different stages of development in both acupuncture and medication groups. In comparison with the control group, serum FSH and E2 levels as well as ovarian Akt and p-Akt protein expression were considerably decreased (P<0.05, P<0.01), and serum LH, T and FINS contents and HOMA-IR were significantly increased in the model group (P<0.01). Following the intervention, the decreased serum FSH and E2 levels as well as ovarian Akt and p-Akt protein expression, and the increased serum LH, T and FINS contents and HOMA-IR were remarkably reversed in both acupuncture and medication groups (P<0.05，P<0.01), without significant differences between the two groups in the levels of those serum and ovary indexes (P>0.05)．. Acupuncture intervention can regulate the levels of reproductive endocrine hormones and reduce insulin resistance in PCOS-IR rats, which may be related to its effect in up- regulating the expression of p-Akt protein in the ovaries.

Effects of apoC1 genotypes on the hormonal levels, metabolic profile and PAF-AH activity in Chinese women with polycystic ovary syndrome

BackgroundElevated serum levels of apolipoprotein (apo) C1 may be an early protein marker of metabolic abnormality in women with polycystic ovary syndrome (PCOS). It is not clear, however, whether there are any relationships between the apoC1 rs4420638A/G and -317deletion (H1)/insertion (H2) polymorphisms and PCOS. We investigated the relationship between these two variants and the risk of PCOS, evaluated the genotypic effects on clinical, hormonal and metabolic indexes and plasma platelet-activating factor acetylhydrolase (PAF-AH) activity, and defined the association of apoC1 gene variants with apoE ε2/ε3/ε4 polymorphisms.MethodsThis is a cross-sectional study of 877 women with PCOS and 761 controls. The apoC1 rs4420638A/G genotype was determined by a Taqman real-time PCR allelic discrimination assay. The apoC1–317H1/H2 and apoE ε2/ε3/ε4 genotypes were measured using PCR and restriction fragment length polymorphism analysis. The clinical, hormonal and metabolic parameters and PAF-AH activity were measured.ResultsThe frequencies of apoC1 rs4420638A/G and -317H1/H2 genotypes and alleles were similar between PCOS and control groups (P > 0.05). However, the rs4420638 G allele was related to increased serum luteinizing hormone, cholesterol and apoB levels, and the ratio of apoB to apoA1 (P < 0.05), and the -317H1H1 genotype was associated with a higher acne grade score and a higher ratio of apoB-PAF-AH to H-PAF-AH activity (P < 0.05) in patients with PCOS. We also demonstrated that the apoC1 rs4420638A/G and -317H1/H2 gene variants existed in moderate to reasonably high linkage disequilibrium with apoE ε2/ε3/ε4 polymorphisms in Chinese women.ConclusionThe apoC1 rs4420638A/G and -317H1/H2 gene variants might be involved in endocrine abnormalities of reproductive axis, metabolic abnormalities and chronic inflammation in PCOS, although no association was observed between the apoC1 genetic variants and the risk of PCOS in Chinese women.