Serum Gonadotropin-releasing Hormone Research Articles

Phoenixin's Influence on HPG Axis and Inflammation in Elite Ice Hockey Athletes: A Cross-Sectional Analysis.

The neuropeptide phoenixin (PNX) may be involved in regulating the hypothalamic-pituitary-gonadal (HPG) axis and inflammatory responses. This study aims to investigate the role of PNX in the regulation of HPG axis function in ice hockey players and its impact on body composition. This cross-sectional study included 65 male ice hockey players aged 18-22, divided into untrained, non-elite athlete, and elite athlete groups. Body composition was assessed using bioelectrical impedance analysis, and plasma levels of PNX, gonadotropin-releasing hormone (GnRH), luteinizing hormone (LH), testosterone, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-10 (IL-10) were measured by enzyme-linked immunosorbent assay. Elite ice hockey players exhibited significantly higher lower limb and core skeletal muscle mass, skeletal muscle index, and testosterone levels, aligning with the high-intensity intermittent nature of hockey training. Compared to the non-training group, ice hockey training groups showed elevated levels of PNX, GnRH, testosterone, and TNF-α, along with reduced levels of LH and IL-6. PNX concentration positively correlated with lean body mass, skeletal muscle mass, skeletal muscle index, serum GnRH, and testosterone levels, and negatively correlated with serum LH and IL-6 levels. In conclusion, PNX may enhance skeletal muscle mass in ice hockey players, particularly in the lower limbs and core muscles, by promoting HPG axis activity while inhibiting inflammatory responses and reducing HPG axis suppression. These findings provide new insights into the physiological adaptation mechanisms of ice hockey players, potentially aiding in the optimization of training strategies and improvement of athletic performance.

Active immunization with a novel recombinant GnRH vaccine inhibits reproductive function in male goats.

Gonadotropin-releasing hormone (GnRH) vaccines have been widely used to effectively inhibit gonadal development and reproductive function. To improve the immunogenicity of GnRH, we developed and evaluated the effects of GnRH6-kisspeptin-CRM197 immunization on the reproductive function in male goats. Thirty 3-month-old male goats (n = 30) were randomly assigned to control, surgical, and immunized groups. The immunized group received a 2 mL injection of the GnRH6-kisspeptin-CRM197 with a booster administered four weeks later. The control group was administered a white oil adjuvant. Blood samples were collected at regular intervals, and at week 20, the animals were euthanized for tissue collection. Serum antibody titers and testosterone levels were measured using ELISA and CLIA, respectively. Testicular parameters and histology were evaluated. The mRNA levels of reproductive-related genes in the HPG axis were measured using RT-qPCR. The results showed that the immunized goats had significantly increased serum GnRH and kisspeptin antibodies (P < 0.05) but decreased testosterone concentrations (P < 0.05) compared to the control group. Testicular size and histology were significantly affected in the immunized group, with notable reductions in testicular weight and dimensions (P < 0.01), and evidence of vacuolar degeneration and suppressed sperm production. The mRNA levels of FSHβ and LHβ in the pituitary, as well as FSHR, LHR, 3βHSD, and 17βHSD in the testis, were significantly lower in the immunized group compared to controls (P < 0.05). These findings suggest that GnRH6-kisspeptin-CRM197 is a safe antigen and a promising immunocastration vaccine with enhanced efficacy.

Active immunization with recombinant GnRH6-CRM197 inhibits reproductive function of male rats

Gonadotropin-releasing hormone (GnRH) vaccines have been successfully used for the inhibition of gonadal development and function, but current GnRH-based vaccines often present variability in the response. Cross-reactive material 197 (CRM197) has been used as carrier molecules to enhance an immune response to associated antigens. So, the synthetic mammalian tandem-repeated GnRH hexamer (GnRH6) gene was integrated into the expression plasmid pET-21a. Recombinant GnRH6-CRM197 protein was subsequently overexpressed in Escherichia coli strain BL21 and purified through Nickel column affinity chromatography and the antigenicity and biological effects of GnRH6-CRM197 were evaluated in rats. Sixteen 4-month-old adult male rats were randomly divided into two groups: the GnRH6-CRM197 group (n = 8) and the control group (n = 8). The GnRH6-CRM197 group rats were subcutaneously immunized with 100 μg of GnRH6-CRM197, administered thrice at 2-week intervals with GnRH6-CRM197.The control group received only a white oil adjuvant. Following the initial immunization, the weights of animals were recorded, and blood samples were collected from the orbital sinus at 4, 4.5, 5, 5.5, 6, 6.5, and 7 months. Serum antibody titers and testosterone concentrations were quantified using ELISA and CLIA, respectively. Additionally, testicular tissues were collected for morphological examination. The results revealed a significant increase in serum GnRH antibody titers (p < 0.05), but a significant decrease in serum testosterone concentrations (p < 0.05), and the weight, length, width, and girth of the testis, and the number of spermatogonia cells, spermatocytes, and sperm cells in the immunized rats. Furthermore, seminiferous tubules revealed significant atrophy and no sperm were observed in the immunized animals. Thus, GnRH6-CRM197 may be an effective antigen and a potential immunocastration vaccine.

Impact of Short-Term Arsenic Exposure on Hormonal Balance and Fibrosis in Sprague Dawley Rats

Objective: To evaluate the toxic effects of arsenic exposure on the female reproductive tract and endocrinesystem.Study Design: A laboratory-based experiment.Place and Duration of Study: The study was carried out at the Laboratory of the National Institute of Health(NIH) Islamabad, Pakistan from 1st January 2019 to 4th January 2020.Methods: non-probability convenient sampling technique was employed and 60 female healthy SpragueDawley rats were included in the study at the National Institute Islamabad. Animals were randomly allocatedinto control and experimental groups (n=30 each). In the first week, animals were acclimatized to the animalhouse of the College of Physicians and Surgeons Pakistan. In the second week, the control and experimentalgroups were given 10 ml of distilled water by oral gavage daily and 4μg of sodium arsenite dissolved in 10 ml ofdistilled water daily for two weeks respectively. After the experimental period, animals were taken to theanimal lab of the National Institute of Health Islamabad for blood sampling and tissue dissection. Rats wereeuthanized by chloroform inhalation and blood was drawn by single intracardiac puncture and stored in clotactivator vials at 4 0C for further workup. The female reproductive tract was dissected, and the tissue was stored in 10 % formalin for histological studies. Serum Gonadotropin-releasing hormone (GnRH) levels were assessed via the Enzyme-Linked Immunosorbent Assay (ELISA) method. Whereas histological effects of arsenic were observed under a microscope by tissue processing and staining with Masson trichrome stain. Mean ± SD of serum GnRH levels and percentage of fibrosis in uterine tissue of control and experimental groups wereevaluated by the software Statistical Package for Social Sciences version 22.0. Comparison of means evaluatedby Student T-test. The significant p-value was ≤ 0.05.Results: The experimental group showed significant elevation of serum GnRH levels and fibrotic changes withhigh collagen deposition (p ≤ 0.05).Conclusion: Exposure to arsenic via contaminated drinking water is disruptive for the female reproductive tractand endocrine system. How to cite this: Mir A, Mir UZ, Hussain W, Irfan A, Butt HA, Sarwar U. Impact of Short-Term Arsenic Exposure on Hormonal Balance and Fibrosis in Sprague Dawley Rats. Life and Science. 2024; 5(2): 187-192. doi: http://doi.org/10.37185/LnS.1.1.64

Characterization of reproductive hormones and related gene expression in the hypothalamus and pituitary gland in the egg-laying interval in White King pigeon

The egg-laying interval (LI) directly reflects the laying performance of breeding pigeons, influenced by reproductive hormones. This study aimed to assess reproductive hormone levels in serum and the expression of related genes and their receptors in the hypothalamus and pituitary gland in 4 stages: first (LI1), third (LI3), fifth (LI5), and seventh (LI7) days. The results showed that serum gonadotropin-releasing hormone (GnRH) level decreased from LI1 to LI7 (P < 0.01) and peaked in LI1. The serum follicle-stimulating hormone (FSH) and luteinizing hormone (LH) levels stayed at high levels from LI1 to LI5. The FSH level decreased slightly from LI5 to LI7 (P > 0.05), but the LH level decreased rapidly (P < 0.01). The prolactin (PRL) levels significantly increased in LI5 (P < 0.01) compared with LI1 and then stayed at a high level. The GnRH1 expression in the hypothalamus had no significant change in LI (P > 0.05). However, the GnRHR first decreased from LI1 to LI3 (P < 0.05) and then increased. The FSH mRNA level in the pituitary gland decreased from LI1 to LI3 and slightly increased in LI5 (P > 0.05). The change pattern of FSHR was similar to that of FSH and peaked in LI5 (P < 0.05). The LH expression level was the highest in LI5 and significantly higher than that in LI3 and LI7 (P < 0.05). However, the LHR mRNA level decreased in LI (P < 0.05). The expression patterns of PRL and PRLR were similar; they were upregulated in LI and peaked in LI7 (P < 0.01). The expression pattern of GnRHR was similar to that of FSH, LH, and FSHR, suggesting the critical role of GnRHR in LI. Furthermore, the expression levels of these genes peaked in LI5, closely correlating with the maturation of the first largest follicle in pigeons. PRL-PRLR signaling inhibited GnRH activity to promote ovulation. This study provided a basis for further investigating the molecular mechanisms underlying the regulation of reproduction in pigeons.

Discovery of microglia gonadotropin‑releasing hormone receptor and its potential role in polycystic ovarian syndrome.

Hypothalamic inflammation is a pathophysiological basis of polycystic ovarian syndrome (PCOS), while overactivated and/or excess M1 polarized microglia are considered to be the main reason for the occurrence of hypothalamic inflammation. Therefore, invitro and invivo experiments were performed to assess the relationships between microglia‑mediated inflammatory reactions and endocrine functions in the PCOS hypothalamus. The expression of gonadotropin‑releasing hormone (GnRH) receptor (GnRHR) was demonstrated in hypothalamic microglia, and it was found that low concentration, GnRH agonist, leuprolide acetate accelerated the expression of M2 polarization marker CD206, while high concentration leuprolide acetate increased the expression of M1 polarization marker CD86 invitro. Furthermore, aerobic exercise not only reduced the levels of serum testosterone, luteinizing hormone and GnRH and the amount of overactivated microglia, but also increased the number of M2 microglia in the hypothalamus of letrozole‑induced PCOS rats. In combination, these results not only demonstrated the expression of GnRHR in hypothalamic microglia, but also demonstrated that GnRH can induce microglial polarization, while aerobic exercise may improve the microglia‑mediated inflammatory reaction by reducing the expression of GnRHR in the hypothalamic microglia of PCOS rats.

Adolescence is a sensitive period for acrylamide-induced sex hormone disruption: Evidence from NHANES populations and experimental mice

Acrylamide (AA) is widely contaminated in environment and diet. However, the association of AA and sex hormones has rarely been investigated, especially in adolescents, a period of particular susceptibility to sex hormone disruption. In this study, survey-weighted multivariate linear regression models were conducted to determine the association between AA Hb biomarkers [HbAA and glycidamide (HbGA)] and sex hormones [total testosterone (TT) and estradiol (E2)] in a total of 3268 subjects from National Health and Nutrition Examination Survey (NHANES) 2013–2016 waves. Additionally, adult and pubertal mice were treated with AA to assess the effect of AA on sex hormones and to explore the potential mechanisms. Among all the subjects, significant negative patterns for HbGA and sex hormones were identified only in youths (6–19 years old), with the lowest β being − 0.53 (95% CI: −0.80 to −0.26) for TT in males and − 0.58 (95% CI: −0.93 to −0.23) for E2 in females. Stratified analysis further revealed significant negative associations between HbGA and sex hormones in adolescents, with the lowest β being − 0.58 (95% CI: −1.02 to −0.14) for TT in males and − 0.54 (95% CI: −1.03 to −0.04) for E2 in females, while there were no significant differences between children or late adolescents. In mice, the levels of TT and E2 were dramatically reduced in AA-treated pubertal mice but not in adult mice. AA disturbed the expression of genes in the hypothalamic–pituitary–gonadal (HPG) axis, induced apoptosis of hypothalamus-produced gonadotropin-releasing hormone (GnRH) neurons in the hypothalamus and reduced serum and hypothalamic GnRH levels in pubertal mice. Our study indicates AA could reduce TT and E2 levels by injuring GnRH neurons and disrupting the HPG axis in puberty, which manifested as severe endocrine disruption on adolescents. Our findings reinforce the idea that adolescence is a vulnerable stage in AA-induced sex hormone disruption.

Expression of phoenixin-14 and nesfatin-1 in the hypothalamo–pituitary–gonadal axis in the phases of the estrous cycle

Phoenixin-14 (PNX-14) and nucleobindin 2 (NUCB2)/nesfatin-1 are regulatory neuropeptides expressed in the hypothalamus. These neuropeptides can be effective in hormonal regulation of the hypothalamo–pituitary–gonadal (HPG) axis and reproductive functions. In the present study, the distribution of PNX-14 and NUCB2/nesfatin-1 in the hypothalamus, pituitary, ovary, and uterus tissues during the phases of the estrous cycle in female rats was investigated. Eighteen Wistar Albino rats determined among animals showing regular estrous cycle by vaginal smear method were divided into three groups: proestrus (Group I), estrus (Group II) and diestrus (Group III). Serum gonadotropin-releasing hormone (GnRH), plasma PNX-14, and NUCB2/nesfatin-1 concentrations were the highest, moderate, and lowest in estrus, diestrus, and proestrus phases, respectively. PNX-14 immunoreactivity in the supraoptic and arcuate nuclei of the hypothalamus and NUCB2/nesfatin-1 immunoreactivity in the paraventricular nuclei were particularly evident in the estrus phase. These neuropeptide immunoreactivities were decreased in different cells of anterior pituitary during proestrus compared with those during estrus and diestrus. PNX-14 immunoreactivity in the ovary, especially during the estrus phase, was diffuse and intense in the granulosa and luteal cells and oocytes, and it was few and weak in theca cells. In addition, NUCB2/nesfatin-1 immunoreactivity was abundant and strong in granulosa and luteal cells, theca and interstitial cells, and oocytes during estrus. In the estrus phase, PNX-14 immunoreactivity was strong in the glandular epithelial cells and stromal cells of the endometrium, also NUCB2/nesfatin-1 immunoreactivity was strong in the epithelial and glandular epithelial cells. As a result, when the estrous cycle was evaluated, it was concluded that the changes in the distribution of PNX-14 and NUCB2/nesfatin-1 at all phases were related to GnRH and that these neuropeptides showed the highest immunoreactivity especially in the HPG axis and uterus tissues of estrus rats.

Diet and exercise interventions reduce serum asprosin and the corresponding hypothalamic– pituitary–gonad-axis dysfunction in obese men

Background: Asprosin (ASP) is a recently discovered adipocyte factor that participates in glucose metabolism and inflammatory reactions. Recent findings suggest that it may be involved in the regulation of sex hormone secretion in the hypothalamic-pituitary-gonad (HPG) axis, but no studies have been reported in related populations. The purpose of this study was to evaluate the changes in serum ASP levels in healthy men and obese men, as well as before and after exercise weight loss, and to investigate male hypogonadism, insulin resistance, inflammatory response, and relationships induced by ASP and obesity. Methods: Thirty-eight young male volunteers were recruited and divided into a normal group (n = 20) and an obese group (n = 18) according to their body mass index. Fourteen of the obese men underwent a 14-week exercise and diet intervention (first 8 weeks of aerobic exercise at 60%–70% HRmax for 30–50 min/4 days a week). Beginning at week 9, the intensity was increased to 75% HRmax. Participants in the obese groups maintained a calorie-restricted diet throughout the study period. Results: Serum ASP levels in the obese group were significantly higher than those in the normal group, and serum gonadotropin-releasing hormone (GnRh), luteinizing hormone (LH), and testosterone (T) levels were decreased. After 14 weeks of exercise and diet intervention, serum ASP decreased significantly, the levels of body weight, lean body weight, body fat rate, fasting insulin (FINS), homeostatic model assessment for insulin resistance, TNF-α, IL-6, and IL-1β decreased significantly, and the serum GnRH, LH, and T levels increased significantly. ASP was positively correlated with body weight, body fat percentage, FINS, tumor necrosis factor (TNF)-α, interleukin (IL)-6, and IL-1β and negatively correlated with relative lean body weight and serum LH and T levels. Conclusion: The serum ASP levels were increased in obese men compared with those of normal weight individuals, resulting in a chronic inflammatory reaction, high serum insulin, and HPG axis injury. Fourteen weeks of exercise and diet intervention effectively alleviated this phenomenon. It has been speculated that ASP might regulate male reproductive function by regulating the inflammatory response and insulin sensitivity.

Effects of dietary supplementation of N-carbamylglutamate on the haematology parameters, secondary sexual characteristicsand testicular gene expression in roosters.

A total of 288 11-week-age roosters were used to evaluate the effects of dietary supplementation of N-carbamylglutamate (NCG) on reproductive traits and testicular gene expression. The experimental periods were 12 weeks. All birds were randomly assigned to 4 treatments with 6 replicates per treatment and 12 birds per replicate. Dietary conditions were based on a basal diet and supplemented with 0%, 0.08%, 0.12%, or 0.16% NCG to form C, N1, N2and N3 groupsrespectively. Dietary supplementation of NCG had positive effects on the seminiferous tubule parameters, serum gonadotropin-releasing hormone and testosterone levelsand the secondary sexual characteristics. Transcriptomics analysis was performed on the testicular tissues between C and N3 groups at the 16-week-age. Genes were mainly enriched in nine pathways, such as cytochrome P450 exogenous metabolism, drug metabolism, steroid hormone synthesisand glutathione metabolism, in which the ZP4 gene, cytochrome P450 family member 11A1and other genes involved in the maintenance of gonadal function, steroid hormone biosynthesis and metabolism, and soforth, exist differences in expression levels. In summary, dietary supplementation of NCG had positive effects on the reproductive traits of roosters. NCG supplementation improved the development of reproductive traits of roosters by regulating the genes expression in testicular tissues and thus improved the synthesis of reproductive hormones in vivo.

Studies on testicular ultrastructural and hormonal changes in type-2 diabetic rats treated with highly active antiretroviral therapy conjugated silver nanoparticles

AimThis study investigated the impact of highly active antiretroviral therapy (HAART) loaded silver nanoparticles (AgNPs) as HAART-AgNPs on the sperm count, viability, serum hormonal profile, insulin-like growth factor I (IGF-1), and testicular ultrastructure. MethodsThirty-six adult male Sprague-Dawley rats were allocated into diabetic and non-diabetic groups (n = 18). The rats in the diabetic group were induced experimental type 2 diabetes using fructose and streptozotocin (frt-STZ). Animals in both groups were subdivided into three groups each, A-C and DF (n = 6), and received distilled water, HAART, and HAART-AgNP, respectively. FindingsTreatment with HAART-AgNP displayed a significant increase (p < 0.05) in serum gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and testicular IGF-1 in diabetic rats. Also, electron microscopy revealed ameliorated testicular ultrastructure upon administration of HAART-AgNP in diabetic rats that were previously marked with architectural and cellular alterations. In addition, treatment with HAART-AgNP significantly reduced (p < 0.05) the blood glucose levels of diabetic rats. In contrast, the treatment of non-diabetic rats with HAART caused a significant decrease (p < 0.05) in the sperm count, serum GnRH, and testicular IGF-1, however, this treatment induced ultrastructural changes and a significant increase (p < 0.05) in serum testosterone levels in diabetic and non-diabetic rats. SignificanceThis study has demonstrated the beneficial impact of HAART-AgNP on the hypothalamic-pituitary-gonadal axis, IGF-1, and testicular architecture in male frt-STZ induced diabetic rats. This nanoconjugate could be a potential nano-drug candidate to cater for testicular dysfunction and metabolic derangements while managing HIV-infected male individuals.

Association between diabetes and IgM antibodies against endogenous gonadotropin-releasing hormone in serum. A meta-analysis

Background: Some studies reported a positive association between diabetes and GnRH IgM antibodies. We aimed to pool the published data to evaluate if there is a link between them or not.

Dietary supplementation with Forsythia suspensa extract during late gestation improves reproductive performance, colostrum composition, antioxidant status, immunoglobulin, and inflammatory cytokines in sows and newborn piglets

This study aimed to evaluate the effects of dietary Forsythia suspensa extract (FSE) supplementation during late gestation on reproductive performance, colostrum composition, nutrient digestibility, antioxidant status, immune status, and inflammatory cytokines in sows and newborn piglets. Forty healthy Yorkshire × Landrace gestating sows (average parity of 3.28 ± 0.61; average body weight of 240.16 ± 6.81 kg) were randomly assigned to 2 dietary treatments with 20 sows per treatment based on body weight, back fat thickness and parity. From d 85 of gestation to farrowing, sows were fed one of two diets including a control diet (CON) and a FSE diet (CON + 100 mg/kg FSE). The results showed that sows fed FSE had increased (P < 0.05) litter birth weight, enhanced (P < 0.05) fat and protein contents in colostrum, and decreased (P < 0.05) rate of stillbirth compared with CON. Dietary FSE supplementation increased (P < 0.05) the apparent total tract digestibility of dry matter, crude protein, ether extracts, gross energy, and organic matter in gestating sows compared with CON. Dietary FSE supplementation in sows increased (P < 0.05) the contents of total antioxidant capacity and superoxide dismutase, decreased (P < 0.05) the content of malondialdehyde in colostrum, and increased (P < 0.05) the contents of catalase and glutathione peroxidase in serum of their offsprings compared with CON. Additionally, sows fed FSE showed higher immunoglobulin A (P < 0.05) in serum of their offsprings and a trend for higher immunoglobulin M (P = 0.08) in colostrum compared with CON. Sows fed FSE showed increased (P < 0.05) content of glucose in colostrum and decreased (P < 0.05) content of urea in serum of their offsprings compared with CON. Sows fed FSE had decreased (P < 0.05) content of interleukin (IL)-6 and increased (P < 0.05) content of IL-10 in serum and colostrum compared with CON. Sows supplemented with FSE had decreased (P < 0.05) contents IL-6 and IL-8 in serum of their offsprings compared with CON. Sows supplemented with FSE also had increased (P < 0.05) content of gonadotropin-releasing hormone in serum of their offsprings compared with CON. In conclusion, dietary supplementation with FSE in sows during late gestation could improve litter birth weight, colostrum fat and protein contents, nutrient digestibility, antioxidant status, immunoglobulin, inflammatory cytokines in sows and newborn piglets. Moreover, FSE could also potentially modulate reproductive hormones in sows.

Protective Effects and Possible Mechanisms of Actions of Bushen Cuyun Recipe on Diminished Ovarian Reserve Induced by Cyclophosphamide in Rats.

BackgroundsDiminished ovarian reserve (DOR) contributes significantly to female infertility. Bushen Cuyun Recipe (BCR, Tradename Yueliang Yin), a product marketed in China, has shown effects in the treatment of female infertility in clinical practices of traditional Chinese medicine (TCM). In this study, we aimed to investigate the chemical compositions of BCR and its efficacy based on scientific evidence and pharmacological mechanisms in DOR treatments.MethodsThe chemical compositions of BCR were determined by the UHPLC-LTQ-Orbitrap MS method. DOR was induced in a rat model by intraperitoneal injection of cyclophosphamide (CTX) 90 mg/kg once. After the CTX treatment for 14 days, rats were intragastrically administrated deionized water, dehydroepiandrosterone (DHEA), or BCR in low, middle, and high doses for 30 days. Ovarian index, ovarian morphology, follicle number, and anti-Müllerian hormone (AMH) in serum were determined to assess the effects of BCR. To investigate possible action mechanisms, network pharmacological analysis was used to predict possible pathways in the effects of BCR on female infertility. In experimental studies, the contents of hormones in the hypothalamic-pituitary-ovarian axis (HPOA, including estradiol (E2), follicle-stimulating hormone (FSH), and gonadotropin-releasing hormone (GnRH)) and pyroptosis-related proteins, including gasdermin D (GSDMD), caspase-1, and interleukin-18 (IL-18), in ovarian were detected by ELISA, immunofluorescence and Western blot.ResultsChemical studies revealed a total 84 components in BCR, which included 43 flavonoids, 13 triterpenoids, 11 phenolic acids, 8 alkaloids, 1 coumarin, 1 anthraquinone, and 7 other components. After treatments with BCR, the ovarian morphology, ovarian index, estrous cycle, growing follicles and corpus luteum from last ovulation, and serum AMH in DOR rats were significantly improved. Network pharmacological analysis suggested that the NOD-like receptor signaling pathway ranked No. 1 among the mechanisms by which BCR affects female infertility. Experimental results demonstrated that the content of serum FSH in DOR rats was significantly decreased and the contents of serum GnRH and E2 were significantly elevated after BCR treatment and that the elevated level of GSDMD, caspase-1, and IL-18 was significantly reversed in BCR-treated rats.ConclusionsThe chemical compositions of BCR were first identified in the present study. BCR was demonstrated to show protective effects on DOR. The possible mechanisms of BCR on DOR might be mediated by regulating gonadal hormones of the HPOA and protecting granulosa cells in ovary against pyroptosis.

The mechanism of Oatp1a5-mediated microcystin-leucine arginine entering into GnRH neurons

Microcystin-leucine arginine (MC-LR) enters into gonadotropin-releasing hormone (GnRH) neurons and induces decline of serum GnRH levels resulting in male reproductive toxicity via hypothalamic-pituitary-testis axis. The organic anion transporting polypeptide 1a5 (Oatp1a5) is a critical transporter for the uptake of MC-LR by GnRH neurons. However, the underlying molecular mechanisms of the transport process are still elusive. In this study, we found that the transmembrane domains 2, 8, and 9 played important roles in transporting function of Oatp1a5. In addition, our data demonstrated that N-linked glycosylation was involved in the transport of MC-LR by Oatp1a5. Moreover, we showed that N-linked glycosylation sites Asn483 and Asn492 were vital for the transport function of Oatp1a5. In summary, the study furthered our understanding of mechanisms that the uptake of MC-LR by GnRH neurons and laid a theoretical foundation for preventing MC-LR from injuring male reproductive health.

Role of edaravone in alleviating prognosis of rats following traumatic brain injury

Objective To investigate the effects of edaravone on improving the prognosis of TBI rats. Methods A total of 150 SD male rats were divided into normal control group (10 rats), TBI group (70 rats) and edaravone group (70 rats). In the edaravone treatment group, the rats were injected intraperitoneally once a day continously for 2 weeks with the injection dose of 5.4 mg·kg-1·d-1. At 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 1 week and 2 weeks after injury, the neurobehavioral and motor function scores of rats were monitored respectively, with 10 rats monitored at each time point. Serum and cerebrospinal fluid samples were collected and the levels of β-endorphin and gonadotropin-releasing hormone (GnRH) were determined by radioimmunoassay (RIA). Results In the edaravone group, the neurobehavioral and motor function scores were higher than those of the TBI group at 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 1 week and 2 weeks after injury. At 48 hours after injury, the neurobehavioral scores of the TBI group and the edaravone treatment group were (8.2±0.9)points and (10.3±0.7)points, respectively (P<0.05), and the motor function scores were (5.9±1.0)points and (6.9±1.2)points respectively (P<0.05). Meanwhile, the contents of β-endorphin in blood and cerebrospinal fluid of the normal control group were (50.2±9.5)pg/ml and (16.2±2.8)pg/ml, and the contents of GnRH were (75.2±11.2)pg/ml and (36.2±10.8)pg/ml, respectively.The levels of β-endorphin and GnRH in serum and cerebrospinal fluid were significantly increased at 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 1 week and 2 weeks after injury. The levels of β-endorphin and GnRH in the edaravone group were lower than those of TBI group. At 72 hour after injury, the levels of β-endorph in serum in TBI group and edaravone group were (165.2±8.5) pg/ml and (109.5±6.3)pg/ml respectively (P< 0.05), and the levels of β-endorph in cerebrospinal fluid were (63.3±3.1)pg/ml and (38.2±2.3)pg/ml respectively (P<0.05). At 72 hour after injury, the levels of GnRH in serum in TBI group and edaravone group were (203.7±17.1)pg/ml and (110.4±19.2)pg/ml respectively (P<0.05), and the levels of GnRH in cerebrospinal fluid is (153.0±13.4)pg/ml and (93.2±10.5)pg/ml respectively (P<0.05). Conclusion During acute and recovery periods after TBI, continuous treatment with edaravone can obviously reduce the levels of β-endorphin and GnRH, which is beneficial to alleviate the secondary brain injury after TBI in rats, promote the recovery of nerve and function, and improve the prognosis. Key words: Brain injuries; β-Endorphin; Gonadotropin-releasing hormone; Edaravone

Evaluation of the serum sex hormones levels and alkaline phosphatase activity in rats’ testis after administering of berberine in experimental varicocele

Current study was aimed to investigate the protective effect of berberine (BB) on the serum gonadotropin-releasing hormone (GnRH), follicle-stimulating hormone (FSH), luteinizing hormone (LH), inhibin B (INHB), testosterone (T) and alkaline phosphatase (Alk-p) activity in the testis of experimental varicocele-induced animals. For the current objective, 30 mature-male Wistar rats were randomly divided into control (n = 6 rats), control-sham (n = 6 rats) and experimental groups (n = 18 rats). The animals in the experimental groups were undergone experimental varicocele and simple laparotomy was conducted in control-sham group. 60 days after varicocele (VCL) induction the experimental group subdivided into: non-treated VCL-induced and 50 mg/kg and 100 mg/kg BB-treated groups (intra-peritoneally). Following 60 days, the animals were euthanized and serum levels of testosterone and testicular activity of alkaline phosphatase were measured. Non-treated VCL-induced animals indicated a significant (P < 0.05) reduction in serum levels of T and INHB and a remarkable (P < 0.05) increase in GnRH, FSH, LH and Alk-p activity compared to control and control-sham groups. Insignificant changes were found between control and control-sham groups. Meanwhile, each BB administered group showed a remarkable (P < 0.05) increase in serum levels of T and INHB and a significant (P < 0.05) decrease in GnRH, FSH, LH and alkaline phosphatase activity in testis tissue. According to the current findings, BB by increasing serum levels of testosterone and INHB increases the testicular endocrine capacity and protects Leydig cell against inflammatory and oxidant injury of varicocele. In addition, BB by inhibiting GnRH, FSH, LH and alkaline phosphatase activity, regulate the levels of serum sex hormones in experimental varicocele and reduces varicocele-induced inflammatory reactions.

MiR-664-2 impacts pubertal development in a precocious-puberty rat model through targeting the NMDA receptor-1†.

Precocious puberty (PP) commonly results from premature activation of the hypothalamic-pituitary-gonadal axis (HPGA). Gonadotropin-releasing hormone (GnRH) is the initial trigger for HPGA activation and plays an important role in puberty onset. N-methyl-D-aspartate (NMDA) can promote pulsatile GnRH secretion and accelerates puberty onset. However, the mechanism of N-methyl-D-aspartate receptors (NMDARs) in PP pathogenesis remains obscure. We found that serum GnRH, luteinizing hormone (LH), follicle-stimulating hormone (FSH), estrogen (E2) levels, hypothalamic NMDAR1, and GnRH mRNA expression peaked at the vaginal opening (VO) day. Next, the hypothalamic NMDAR1 mRNA and protein levels in rats treated with danazol, a chemical commonly effecting on the reproductive system, were significantly increased at the VO day (postnatal day 24) compared to controls, accompanied by enhanced serum GnRH, LH, FSH, and E2 levels. Further, microRNA-664-2 (miR-664-2) was selected after bioinformatics analysis and approved in primary hypothalamic neurons, which binds to the 3'-untranslated regions of NMDAR1. Consistently, the miR-664-2 expression in hypothalamus of the Danazol group was decreased compared to Vehicle. Our results suggested that attenuated miR-664-2 might participate in PP pathogenesis through enhancing the NMDAR1 signaling.

Warm needle moxibustion, mild moxibustion and electroacupuncture interventions have an anti-aging effect possibly by regulating hypothalamus-pituitary-testis axis in aged male rats

To observe the effect of warm needle moxibustion (WNM), mild moxibustion and electroacupuncture (EA) of "Shenshu" (BL23) and "Guanyuan" (CV4) on serum gonadal hormone levels and related gene expression in the hypothalamus and testis in aged rats, so as to explore their underlying mechanisms in anti-aging. Forty male SD male rats were randomly divided into aged control, WNM, mild moxibustion, EA and medication groups (n ＝ 8 in each group), and other 8 male young adult SD rats were used as the youth control group. WNM (acupuncture plus moxibustion), mild moxibustion or EA (2 Hz /100 Hz, 1 mA) was separately applied to CV4 and bilateral BL23 for 15 min, once daily, 5 days a week for 8 weeks. Rats of the medication group received subcutaneous injection of testosterone propionate injection (7 mg/kg), once every 3 days, and those of the aged and youth groups received the same dose of 0.9% NaCl solution for 8 weeks. The levels of serum testoste-rone (T) and gonadotropin-releasing hormone (GnRH) were measured by ELISA. The expression of Kisspeptin (KISS-1), G protein-coupled receptors 54 (GPR54), RFamide-related peptide-3 (RFRP-3) and GPR147 mRNAs in the hypothalamus and GnRH-R, GPR54 and GPR147 mRNAs in the testis tissues were detected by quantitative real-time PCR. The levels of serum T and GnRH, and the expressions of KISS-1, GPR54, RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R and GPR54 mRNAs in the testis in the aged rats were significantly lower than those in the youth rats (P<0.05), while the expression level of GPR147 mRNA of the testis tissue was obviously higher in the aged group than that in the youth group (P<0.05). Follo-wing the treatment, the lowered serum T and GnRH contents, the down-regulated expression levels of hypothalamic KISS-1, GPR54, RFRP-3 and GPR147 mRNAs and testicle GnRH-R and GPR54 mRNAs, as well as the up-regulated testicle GPR147 mRNA expression in the aged rats were reversed in the WNM, mild moxibustion and EA groups (not the medication group except serum T) in comparison with the aged control group (P<0.05). The therapeutic effects of WNM, mild moxibustion and EA were significantly superior to those of medication in up-regulating serum GnRH content and expression levels of KISS-1, GPR54, RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R, GPR54 and GPR147 mRNAs in the testis (P<0.05). No significant differences were found among the WNM, mild moxibustion and EA groups in serum T and GnRH contents, and the expression levels of KISS-1, GPR54, RFRP-3 and GPR147 mRNAs in the hypothalamus and GnRH-R and GPR54 and GPR147 mRNAs in the testis (P>0.05)．. The WNM, mild moxibustion and EA therapies have an anti-aging effect by increasing serum T and GnRH levels in aged rats, which is possibly related to their effects in up-regulating the expression of hypothalamic KISS-1, GPR54, RFRP-3 and GPR147 mRNAs and testicle GnRH-R and GPR54 mRNAs, as well as down-regulating testicle GPR147 mRNA expression. The therapeutic effect of WNM, mild moxibustion and EA is evidently superior to that of medication.

Microcystin-leucine arginine inhibits gonadotropin-releasing hormone synthesis in mice hypothalamus

Microcystin-leucine arginine (MC-LR) causes serum testosterone declines and male reproductive disorders. However, the molecular mechanisms underlying the pathological changes are still unclear. In the present study, we aimed to investigate the toxic effects of MC-LR on gonadotropin-releasing hormone (GnRH) neurons in the hypothalamus. Our results demonstrated that MC-LR could enter GnRH neurons and inhibit GnRH synthesis, resulting in the decrease of serum GnRH and testosterone levels. The inhibitory effects of MC-LR on GnRH synthesis were identified to be associated with activation of the cyclic adenosine monophosphate (cAMP)/protein kinase A (PKA)/cAMP response element-binding protein (CREB)/c-Fos signaling pathway. With miRNA microarray analyses, we found that miR-329-3p was down-regulated most dramatically in MC-LR-treated GT1–7 cells. We then further identified that miR-329-3p regulated PRKAR1A and PRKACB expression and thus influenced GnRH synthesis. This is the first study to explore the molecular mechanism underlying the inhibitory effects of MC-LR on GnRH synthesis in the hypothalamus. Our data have provided a new perspective in the development of diagnosis and treatment strategies for male infertility as a result of dysfunction of the hypothalamic-pituitary-gonadal axis.