Plasma Steroid Levels Research Articles

Prevention of chemotherapy-induced premature ovarian insufficiency in mice by scaffold-based local delivery of human embryonic stem cell-derived mesenchymal progenitor cells

BackgroundPremature ovarian insufficiency (POI) is one of the most serious side effects of chemotherapy in young cancer survivors. It may not only reduce fecundity but also affect lifelong health. There is no standard therapy for preserving ovarian health after chemotherapy. Recently, administration of embryonic stem cell-derived mesenchymal progenitor cells (ESC-MPCs) has been considered a new therapeutic option for preventing POI. However, the previous method of directly injecting cells into the veins of patients exhibits low efficacy and safety. This study aimed to develop safe and effective local delivery methods for the prevention of POI using two types of bioinspired scaffolds.MethodsFemale mice received intraperitoneal cisplatin for 10 days. On day 11, human ESC-MPCs were delivered through systemic administration using intravenous injection or local administration using intradermal injection and intradermal transplantation with a PLGA/MH sponge or hyaluronic acid (HA) gel (GEL) type of scaffold. PBS was injected intravenously as a negative control. Ovarian function and fertility were evaluated 4 weeks after transplantation. Follicle development was observed using hematoxylin and eosin staining. The plasma levels of sex hormones were measured using ELISA. Expression levels of anti-Müllerian hormone (AMH) and ki-67 were detected using immunostaining, and the quality of oocytes and embryos was evaluated after in vitro fertilization. The estrous cycles were observed at 2 months after transplantation.ResultsThe local administration of human ESC-MPCs using the bioinspired scaffold to the backs of mice effectively prolonged the cell survival rate in vivo. The HA GEL group exhibited the best recovered ovarian functions, including a significantly increased number of ovarian reserves, estrogen levels, and AMH levels and decreased apoptotic levels. Furthermore, the HA GEL group showed improved quality of oocytes and embryos and estrous cycle regularity.ConclusionsHA GEL scaffolds can be used as new delivery platforms for ESC-MPC therapy, and this method may provide a novel option for the clinical treatment of chemotherapy-induced POI.

Effects of paroxetine treatment and its withdrawal on neurosteroidogenesis

Selective serotonin reuptake inhibitors (SSRI) show high efficacy in treating depression, however during treatment side effects, like for instance sexual dysfunction, may appear, decreasing compliance. In some cases, this condition will last after drug discontinuation, leading to the so-called post-SSRI sexual dysfunction (PSSD). The etiology of PSSD is still unknown, however a role for neuroactive steroids may be hypothesized. Indeed, these molecules are key physiological regulators of the nervous system, and their alteration has been associated with several neuropathological conditions, including depression. Additionally, neuroactive steroids are also involved in the control of sexual function. Interestingly, sexual dysfunction induced by SSRI treatment has been also observed in animal models. On this basis, we have here evaluated whether a subchronic treatment with paroxetine for two weeks and/or its withdrawal (i.e., a month) may affect the levels of neuroactive steroids in brain areas (i.e., hippocampus, hypothalamus, and cerebral cortex) and/or in plasma and cerebrospinal fluid of male rats. Data obtained indicate that the SSRI treatment alters neuroactive steroid levels and the expression of key enzymes of the steroidogenesis in a brain tissue- and time-dependent manner. Indeed, these observations with the finding that plasma levels of neuroactive steroids are not affected suggest that the effect of paroxetine treatment is directly on neurosteroidogenesis. In particular, a negative impact on the expression of steroidogenic enzymes was observed at the withdrawal. Therefore, it is possible to hypothesize that altered neurosteroidogenesis may also occur in PSSD and consequently it may represent a possible pharmacological target for this disorder.

The effects of three types of exercise training on steroid hormones in physically inactive middle-aged adults: a randomized controlled trial.

Physical inactivity and ageing are associated with imbalances in anabolic/catabolic steroid hormones, jeopardizing health. We investigated the effects of three types of training on plasma steroid hormone levels in physically inactive, middle-aged adults. A 12-week randomized controlled trial was performed with a parallel-group design. A total of 67 (36 women) middle-aged adults (45-65years old) were randomly assigned to (1) no exercise (control), (2) concurrent training based on the international physical activity recommendations (PAR), (3) high-intensity interval training (HIIT), or (4) HIIT plus whole-body electromyostimulation (HIIT + EMS). The training volume in the PAR group was 150min/week at 60-65% of the heart rate reserve for aerobic training and ~ 60min/week at 40-50% of the one-repetition maximum for resistance training. The training volume in the HIIT and HIIT + EMS groups was 40-65min/week at > 95% of the maximum oxygen uptake in long interval sessions, and > 120% of the maximum oxygen uptake in short interval sessions. Compared to the control group, dehydroepiandrosterone sulfate increased in the PAR, HIIT, and HIIT + EMS groups (~ 14%, ~ 14%, and ~ 20%, respectively; all P < 0.01). Cortisol decreased in the PAR, HIIT, and HIIT + EMS groups (~ -17%, ~ -10%, and ~ -23%, respectively; all P ≤ 0.05). Testosterone increased in the HIIT and HIIT + EMS groups (~ 28%, and ~ 16%, respectively; all P ≤ 0.01). Free testosterone increased in the HIIT and HIIT + EMS groups (~ 30% and ~ 18% respectively; all P ≤ 0.01). No significant increase in sex hormone-binding globulin was observed (P = 0.869). Our findings suggest that HIIT, with or without whole-body EMS, can significantly enhance steroid hormones status in previously physically inactive middle-aged adults. The PAR program led to slight improvements than the HIIT and HIIT + EMS groups despite the application of a higher training volume. NCT03334357 (ClinicalTrials.gov). November 7, 2017 retrospectively registered.

BIOCHEMICAL, HISTOPATHOLOGICAL, AND GENETIC CHARACTERIZATION OF POSTURE RESPONSIVE AND UNRESPONSIVE ALDOSTERONE-PRODUCING ADENOMAS

Objective: Aldosterone-producing adenoma (APA) accounts for ∼30% of primary aldosteronism. Based on responses of aldosterone to upright posture, APA can be divided into posture-responsive and –unresponsive forms. Somatic gene mutations in KCNJ5, CACNA1D, ATP1A1, or ATP2B3 have been reported to exist in more than 90% of APAs. We sought to compare the biochemical, histopathological, and genetic characterization between these two types of APAs. Design and method: Posture-responsive was defined as showing a rise in plasma aldosterone concentration of at least 50% 2 - 3 h after the assumption of upright posture (10 AM) above the basal level (7 AM) following overnight recumbency. Plasma levels of aldosterone and hybrid steroids (18-oxocortisol and 18-hydroxycortisol) were measured by LC-MS/MS. Histologic analysis and immunohistochemistry for CYP11B2 (aldosterone synthase) and CYP17A1 (17aplha-hydroxylase) were performed on APA tissue sections collected from 24 posture-unresponsive and 17 posture-responsive APA patients. Sanger sequencing of KCNJ5 and gene targeted next-generation sequencing were performed to detect somatic gene mutations. Results: Compared to patients with posture-responsive APA (of which 48.7% were composed of zona glomerulosa (ZG)-like cells and 35.3% showed ZG-zona fasciculata (ZF) mixed appearance), posture-unresponsive APA patients displayed higher (P &lt; 0.01) levels of hybrid steroids, recumbent aldosterone and cortisol, greater (P &lt; 0.05) suppression of aldosterone secretion from the contralateral unaffected adrenal, and larger (P &lt; 0.01) tumors mostly composed of ZF-like cells that had higher (P &lt; 0.01) expression of CYP17A1 (but not of CYP11B2). Of 41 studied APAs, 37 (90.2%) harboured aldosterone-driving somatic mutations (KCNJ5 = 14, 34.1%; CACNA1D = 13, 31.7%; ATP1A1 = 8, 19.5%; and ATP2B3 = 2, 4.9%), including 5 that were previously unreported (3 in CACNA1D and 2 in ATP1A1). Interestingly, 64.7% (11/17) of posture-responsive APAs carried CACNA1D mutations, whereas 54.2% (13/24) of posture-unresponsive APAs harboured KCNJ5 mutations. Conclusions: The excessive production of hybrid steroids by posture-unresponsive APAs may relate to their ZF-like tumor cell composition, resulting in strong expression of CYP17A1 (in addition to somatic gene mutation-driven CYP11B2 expression), thereby allowing production of cortisol which acts as the substrate for CYP11B2-generated hybrid steroids.

The importance of thermoperiod for proper gametogenesis and successful egg and sperm production in meagre (Argyrosomus regius) breeders in aquaculture

We examined the effect of constant water temperature throughout the year on gametogenesis, spawning success and egg/sperm/embryo quality in meagre (Argyrosomus regius). Two broodstocks were exposed to simulated natural photoperiod, andeither attenuated seasonal water temperature (SeasT, 16.4 to 19.6ºC) or relatively constant water temperature (CoT, 19.4 ± 0.6ºC).In the spawning period (May), 4 couples per group were induced to spawn with gonadotropin releasing hormone agonist (GnRHa). Gonadal stage of development, sperm quality parameters and plasma levels of sex steroids were evaluated prior to the GnRHa treatment. Spawning success and egg/sperm quality were examined over the following 4 weeks. Constant temperature did not prevent gametogenesis, but exposure to attenuated seasonal water temperature with the inclusion of winter low temperature was beneficial to both sexes. The mean (±SD) diameter of the largest vitellogenic oocytes prior to GnRHa administration was significantly higher in the SeasT compared to the CoT group (598 ± 27 vs 520 ± 17 μm). Testosterone plasma levels in the females were significantly higher in the SeasT group, but all other hormones were similar in both sexes. SeasT females spawned more consistently with higher relative fecundity, and 24-h embryo survival of the produced eggs. A more pronounced negative effect of constant water temperature was observed in males, since CoT males exhibited a spermiation index of 0 prior to GnRHa treatment, the latter clearly having a beneficial effect over the following 4 weeks. The study demonstrated that meagre do undergo gametogenesis to a significant extent even under constant water temperatures during the year. However, a seasonal thermal regime -even an attenuated one- was necessary for the proper development of the gametes, allowing for the successful spawning induction using the established GnRHa induction protocol.

Profiling of Steroid Metabolic Pathways in Human Plasma by GC-MS/MS Combined with Microwave-Assisted Derivatization for Diagnosis of Gastric Disorders.

Steroid hormones are associated in depth to cellular signaling, inflammatory immune responses, and reproductive functions, and their metabolism alterations incur various diseases. In particular, quantitative profiling of steroids in plasma of patients with gastric cancer can provide a vast information to understand development of gastric cancer, since both sex hormones and glucocorticoids might be correlated with the pathological mechanisms of gastric cancer. Here, we developed a gas chromatography-tandem mass spectrometry-dynamic multiple reaction monitoring (GC-MS/MS-dMRM) method combined with solid-phase extraction (SPE) and microwave-assisted derivatization (MAD) to determine 20 endogenous steroids in human plasma. In this study, MAD conditions were optimized with respect to irradiation power and time. The SPE enabled effective cleanup and extraction for profiling of steroid hormones in human plasma samples. The MAD could improve laborious and time-consuming derivatization procedure, since dielectric heating using microwave directly increase molecular energy of reactants by penetrating through medium. Furthermore, dMRM method provided more sensitive determination of 20 steroids, compared to traditional MRM detection. The limits of quantification of steroids were below 1.125 ng/mL and determination coefficients of calibration curves were higher than 0.9925. Overall precision and accuracy results were below 19.93% and within ±17.04%, respectively. The developed method provided sufficient detection sensitivities and reliable quantification results. The established method was successfully applied to profile steroid metabolism pathways in plasma of patients with chronic superficial gastritis (CSG), intestinal metaplasia (IM), and gastric cancer. Statistical significances of steroid plasma levels between gastric disorder groups were investigated. In conclusion, this method provided comprehensive profiling of 20 steroids in human plasma samples and will be helpful to discover potential biomarkers for the development of gastric cancer and to further understand metabolic syndrome.

Effect of various intensity of exercise on serum prolactin level in untrained male individuals

Background: Exercise and physical health influence the plasma levels of reproductive hormone and prolactin in both sexes. Lower levels of sex hormones, female estrogen, and male testosterone can be associated with higher levels of serum prolactin. Lower male reproductive hormones may lead to exhaustion, decreased sperm count, loss of sexual desire, temporary infertility, and impotence. Aim and Objective: The objective of the study was to see the effect of various intensity of exercise on serum prolactin level in untrained male individuals. Materials and Methods: A total of 20 untrained male individuals asked to perform exercise of various grade (mild, moderate, and severe). Pre-and post-workout blood samples were obtained to determine serum prolactin levels. Serum prolactin levels assessed through prolactin hormone ELISA kit. The data were analyzed by the student Paired t–test using the version 26 “SPSS Statistical Package.” Results: In untrained male subjects, the prolactin levels increased after the various intensity of exercise compare to pre-exercise level and the difference was statistically significant. Conclusion: In males plasma levels of sex hormones not only depends on postexercise serum prolactin levels but also depending on the intensity and duration of the activity and the physical fitness of the individual.

Consumption of sage (Salvia officinalis) promotes ovarian function by stimulating estradiol hormone release and controlling folliculogenesis, steroidogenesis, and autophagy

ObjectivesSalvia officinalis (sage) is widely sold in Saudi Arabia to make a popular herbal tea that women with reproductive disorders can drink daily. Despite the wide use of sage in traditional medicine, to our knowledge, little attention has been given to its effect on female reproduction. Our purpose was to understand the effects of sage extract on ovarian function to potentially shed light on its use in treating female infertility. MethodsForty female Wistar virgin female rats were randomly assigned to four groups, with 10 animals in each group. The control group received distilled water, and the three treatment groups received different concentrations of sage extract: 3.5, 15 or 60 mg/kg during 14 days. Ovarian tissues were cut into 5-μm sections and stained with hematoxylin and eosin. Plasma levels of reproductive steroids were measured using ELISA, and gene expression was examined via RT-PCR. ResultsOur results showed that low doses of sage (3.5 mg/kg and 15 mg/kg) have no effects on serum concentration levels of steroid hormone, the number of growing follicles, and on the transcripts of the steroidogenesis and folliculogenesis genes. In contrast, a high dose of the extract (60 mg/kg) significantly increased estradiol concentration levels. As a result, the number of growing follicles increased, while the number of abnormal follicles significantly decreased. Interestingly, sage extract also affected mRNA levels of genes involved in folliculogenesis and steroidogenesis processes. Two autophagy related genes, LC3 and ATG12, were not affected, but all of the investigated genes, INSL3, CYP17a, LHR, CCND2, IGF1, ESR1, ESR2, ACTB, GDF, and ATG5, significantly increased with the highest dose of sage. ConclusionAs a remedy, sage extract is best used at a high dose when treating female infertility disorders, as sage extract may promote ovarian function by stimulating folliculogenesis and steroidogenesis.

Kisspeptin Influences the Reproductive Axis and Circulating Levels of microRNAs in Senegalese Sole.

Kisspeptin regulates puberty and reproduction onset, acting upstream of the brain–pituitary–gonad (HPG) axis. This study aimed to test a kisspeptin-based hormonal therapy on cultured Senegalese sole (G1) breeders, known to have reproductive dysfunctions. A single intramuscular injection of KISS2-10 decapeptide (250 µg/kg) was tested in females and males during the reproductive season, and gonad maturation, sperm motility, plasma levels of gonadotropins (Fsh and Lh) and sex steroids (11-ketotestosterone, testosterone and estradiol), as well as changes in small non-coding RNAs (sncRNAs) in plasma, were investigated. Fsh, Lh, and testosterone levels increased after kisspeptin injection in both sexes, while sperm analysis did not show differences between groups. Let7e, miR-199a-3p and miR-100-5p were differentially expressed in females, while miR-1-3p miRNA was up-regulated in kisspeptin-treated males. In silico prediction of mRNAs targeted by miRNAs revealed that kisspeptin treatment might affect paracellular transporters, regulate structural and functional polarity of cells, neural networks and intracellular trafficking in Senegalese sole females; also, DNA methylation and sphingolipid metabolism might be altered in kisspeptin-treated males. Results demonstrated that kisspeptin stimulated gonadotropin and testosterone secretion in both sexes and induced an unanticipated alteration of plasma miRNAs, opening new research venues to understand how this neuropeptide impacts in fish HPG axis.

17\u03b1-ethynylestradiol prevents the natural male-to-female sex change in gilthead seabream (Sparus aurata L.)

Exposure to 17α-ethynylestradiol (EE2, 5 μg/g food) impairs some reproductive events in the protandrous gilthead seabream and a short recovery period does not allow full recovery. In this study, spermiating seabream males in the second reproductive cycle (RC) were fed a diet containing 5 or 2.5 μg EE2/g food for 28 days and then a commercial diet without EE2 for the remaining RC. Individuals were sampled at the end of the EE2 treatment and then at the end of the RC and at the beginning of the third RC, 146 and 333 days after the cessation of treatment, respectively. Increased hepatic transcript levels of the gene coding for vitellogenin (vtg) and plasma levels of Vtg indicated both concentrations of EE2 caused endocrine disruption. Modifications in the histological organization of the testis, germ cell proliferation, plasma levels of the sex steroids and pituitary expression levels of the genes coding for the gonadotropin β-subunits, fshβ and lhβ were detected. The plasma levels of Vtg and most of the reproductive parameters were restored 146 days after treatments. However, although 50% of the control fish underwent sex reversal as expected at the third RC, male-to female sex change was prevented by both EE2 concentrations.

Resumption and progression of meiosis and circulating levels of steroids and prostaglandin F 2α of Piaractus mesopotamicus induced by hypophysation with prostaglandin F 2α

We have previously shown an increase on the ovulation rate of pacu, Piaractus mesopotamicus, using hypophysation with prostaglandin F2α (PGF2α) (HPG protocol) rather than hypophysation (H); only, however, a persistent lack of ovulation in some females could not be further explained by PGF2α and steroid plasma levels at time of ovulation. Considering this, here we compared the meiotic process and steroids and PGF2α levels during spawning induction using both protocols. To that, H and HPG were treated with conventional hypophysation (0.6 mg/kg priming and 5.4 mg/kg resolving dose), and the latter received synthetic PGF2α (2 mL/fish) at resolving dose. A negative control (saline solution), a SPG group (receiving only PGF2α) and a PPG group (receiving priming and PGF2α) were included. An initial PGF2α peak between one to four hours post-resolving dose (1-4 hr) occurred in groups receiving PGF2α (HPG, SPG and PPG). Only H and HPG showed a 17α-20β-dihydroxy-4-pregnen-3-one (DHP) peak 4 hr, concomitant with meiosis resumption, and a final PGF2α peak 12 hours. The HPG protocol resulted in higher PGF2α plasma levels for longer periods (1-4 and 12 hr) than H (single peak 12 hr), occasioning in the former higher and concomitant levels of both PGF2α and DHP at 4 h. Considering that DHP is known to induce expression of prostaglandin receptors in fish, the concomitant plasma peaks of DHP and PGF2α may also explain the ovulation rate gain previously published, using HPG protocol, which can be further improved adjusting the priming dose and defining the exact interval of PGF2α ovulatory action in this species.

Biomonitoring the effects of urban-stream waters on the health status of pale chub (Zacco platypus): A comparative analysis of biological indexes and biomarker levels

The objective of this study was to biomonitor the effects of potential environmental pollutants in urban-stream waters, on fish health. Pale chub (Zacco platypus), a dominant species in the Korea urban stream waters, was chosen and biomonitoring indicators for the different spatial characteristics were tailored in an urban watershed. Biological responses including biotic-somatic index as well as gonadal development phase and plasma steroids levels, and the biochemical responses, ethoxyresorufin-o-deethylase (EROD) and acetylcholinesterase (AChE) activities, were measured. No significant difference was observed in the length–weight relationship between the up-stream waters and the down-stream waters. However, changes in the gonad-somatic index (GSI) levels, plasma 17β-estradiol (E2) levels, and mature oocyte frequencies in the female fish collected during the spawning season were observed in the down-stream waters at each monitoring site. Moreover, intersex condition (testis-ova) in the male fish in down-stream waters was recorded, even if it was just one fish. Although no significant difference was observed in the EROD and AChE activities between the up-stream waters and the down-stream waters, changes in the reproductive biomarker levels, including the GSI levels, plasma E2 levels, and gonadal maturation, lead to variable biomonitoring endpoints between the spatial different sites. These results imply that exposure to the down-stream waters can cause reproductive impairment in wild Z. platypus, individual variability in the biological responses further indicate the reproductive health was affected more by the down-stream waters than the up-stream waters. The finding from this study can provide the biomonitoring endpoint on the wild fish health in urban watershed that is crucial to the early risk assessment of its biological impacts. More multi-biomarkers studies reflecting the variation in the biological organization of wild fish and, therefore, the effects of urban-stream waters in the fish health are warranted.

Histomorphological and endocrine assessment of female Arabian carpetshark, Chiloscyllium arabicum (Elasmobranchii: Hemiscylliidae) from the Persian Gulf during annual reproductive cycle.

The present study aimed at assessing the annual reproductive cycle of female Arabian carpetshark, Chiloscyllium arabicum from the Persian Gulf by a macroscopic and microscopic evaluation of the reproductive tract. The annual cycle of gonadal steroids [17β-estradiol (E2), progesterone (P4) and testosterone (T)] was also assessed in this shark. In total, 130 female C. arabicum were collected from the Bahrakan Creek (located northwest of the Persian Gulf) between January 2018 and March 2019. Females were oviparous with an external-type ovary and only one functional ovary. Five sexual maturity stages were recognized based on macroscopic and microscopic evaluation: Immature I (August-October), Immature II (November-January), Mature (February-March), Pregnant (April-May) and Spent (June-July). The structural changes in the oviducts, oviducal glands and uterus throughout the annual reproductive cycle were consistent with their roles in the egg movement, the egg capsule production and sperm storage. The plasma levels of the gonadal steroids were associated with morphological changes in the reproductive tract. E2 showed two detectable peaks during March (close to ovulation) and June (just before mating). P4 and T displayed a peak just before ovulation.

Sex Hormones and Incident Heart Failure in Men and Postmenopausal Women: The Atherosclerosis Risk in Communities Study.

Sex differences exist in heart failure (HF) phenotypes, but there is limited research on the role of sex hormones in HF and its subtypes. To examine the associations of total testosterone, dehydroepiandrosterone sulfate (DHEA-S), and sex hormone-binding globulin (SHBG) with incident HF, HF with preserved ejection fraction (HFpEF), and HF with reduced ejection fraction (HFrEF). Atherosclerosis Risk in Communities (ARIC) study (prospective cohort study). Median follow-up is 19.2 years. General community. 4107 men and 4839 postmenopausal women, with mean age of 63.2 (standard deviation [SD] 5.7) and 62.8 (5.5) years, respectively. Plasma sex hormone levels were measured at visit 4 (1996-1998). Incident HF events were identified through hospital discharge codes and death certificates. The Hazard Ratios for HF associated with 1 SD decrease in log-transformed total testosterone, DHEA-S, and SHBG were 1.10 (95% confidence interval 1.03, 1.17), 1.07 (1.00, 1.15), and 1.04 (0.96, 1.11) in men, and 1.05 (0.99, 1.13), 1.17 (1.09, 1.24), and 0.93 (0.85, 1.01) in women, respectively. The associations between sex hormones with subtypes of HF had similar patterns but were attenuated and became statistically insignificant. In this prospective cohort, lower levels of endogenous testosterone and DHEA-S in men and DHEA-S in postmenopausal women were associated with the development of HF. Similar directions of association in both sexes and both HF subtypes suggest that sex hormones play a role in the development of HF through common pathways regardless of sex.

Impact of Microbiome on Hepatic Metabolizing Enzymes and Transporters in Mice during Pregnancy.

The microbiome and pregnancy are known to alter drug disposition, yet the interplay of the two physiologic factors on the expression and/or activity of drug metabolizing enzymes and transporters (DMETs) is unknown. This study investigated the effects of microbiome on host hepatic DMETs in mice during pregnancy by comparing four groups of conventional (CV) and germ-free (GF) female mice and pregnancy status, namely, CV nonpregnant, GF non-pregnant, CV pregnant, and GF pregnant mice. Transcriptomic and targeted proteomics of hepatic DMETs were profiled by using multiomics. Plasma bile acid and steroid hormone levels were quantified by liquid chromatography tandem mass spectrometry. CYP3A activities were measured by mouse liver microsome incubations. The trend of pregnancy-induced changes in the expression or activity of hepatic DMETs in CV and GF mice was similar; however, the magnitude of change was noticeably different. For certain DMETs, pregnancy status had paradoxical effects on mRNA and protein expression in both CV and GF mice. For instance, the mRNA levels of Cyp3a11, the murine homolog of human CYP3A4, were decreased by 1.7-fold and 3.3-fold by pregnancy in CV and GF mice, respectively; however, the protein levels of CYP3A11 were increased similarly ∼twofold by pregnancy in both CV and GF mice. Microsome incubations revealed a marked induction of CYP3A activity by pregnancy that was 10-fold greater in CV mice than that in GF mice. This is the first study to show that the microbiome can alter the expression and/or activity of hepatic DMETs in pregnancy. SIGNIFICANCE STATEMENT: We demonstrated for the first time that microbiome and pregnancy can interplay to alter the expression and/or activity of hepatic drug metabolizing enzymes and transporters. Though the trend of pregnancy-induced changes in the expression or activity of hepatic drug metabolizing enzymes and transporters in conventional and germ-free mice was similar, the magnitude of change was noticeably different.

Pharmacomicrobiomics of Hepatic Drug Processing Genes During Pregnancy

At present, little is known about how gut microbiome and pregnancy interplay to regulate expression or the activities of drug processing genes (DPGs). The objective of this study was to investigate the effects of gut microbiome on host hepatic DPGs during pregnancy using conventional (CV) and germ‐free (GF) mice. Four groups of C57BL/6 female mice were used: CV non‐pregnant (CVNP, n=6), GF non‐pregnant (GFNP, n=6), CV pregnant (CVP, n=5), and GF pregnant (GFP, n=5) mice. Hepatic transcriptome and proteome for DPGs were profiled using multi‐omics approach. Plasma bile acids and steroid hormone levels were determined using LC‐MS/MS. Cyp activities were measured using hepatic microsome incubations. Overall, RNA‐seq analysis revealed a similar trend of gene regulation by pregnancy, but different magnitudes of change in hepatic DPGs, in CV and GF mice. Of note, the Cyp3a11 mRNA levels were decreased 1.7‐fold and 3.2‐fold by pregnancy in CV and GF mice, respectively. On the contrary, the Cyp3a11 protein abundance was increased 1.2‐fold and 1.6‐fold by pregnancy in CV and GF mice, respectively. The Cyp3a activity was also significantly induced by pregnancy in CV and GF mice; however, the magnitude of induction in CV mice was &gt;5‐fold greater than that in GF mice. Plasma bile acid and steroid hormone levels were significantly impacted by microbiome and pregnancy‐status, respectively, which may contribute to the differential effects of pregnancy in CV and GF mice. This is the first study to demonstrate that the gut microbiome may play a role in altering hepatic drug disposition in pregnancy.Support or Funding InformationThis project was supported by the University of Washington Drug Metabolism, Transport and Pharmacogenomics Research Program (DMTPR)

Disruptive effects of nonylphenol on reproductive hormones, antioxidant enzymes, and histology of liver, kidney and gonads in Caspian trout smolts

The endocrine-disrupting effect of pollutants such as alkylphenols has been considered a major concern during recent years. Although the effects of nonylphenol on the reproductive hormones of fish have been investigated in several studies, the effect of this endocrine disruptor on reproductive hormones of immature fish and salmonid smolts has been less addressed. The present work studied the effects of 1, 10 and 100 μg/l concentrations of nonylphenol on the levels of plasma reproductive hormones and liver antioxidant enzymes as well as on histopathology of reproductive and non-reproductive organs of male and female Caspian brown trout (Salmo trutta caspius) smolts after 21 days of exposure. The results of the present study showed that environmentally relevant concentrations of nonylphenol affected plasma levels of sex steroids; gonadotropins, phosphorus, estradiol to testosterone ratio, and also caused histopathological lesions in liver, kidney and testis tissues of immature Caspian brown trout during smolting. Nonylphenol significantly increased the levels of estradiol in plasma of both male and female smolts exposed to nonylphenol compared with the control groups. Exposure to nonylphenol decreased testosterone and FSH levels in both genders. It has also increased plasma levels of LH in females but did not affect LH levels in male fish. Liver SOD and CAT content was decreased in nonylphenol-exposed smolts. Therefore, the release of this economically valuable and endangered species into the rivers contaminated with nonylphenol should be avoided as it can have significant effects on the development and reproductive function of smolts.

In vivo blockade of ovarian sympathetic activity by Neosaxitoxin prevents polycystic ovary in rats.

A high sympathetic tone is observed in the development and maintenance of the polycystic ovary (PCO) phenotype in rats. Neosaxitoxin (NeoSTX) specifically blocks neuronal voltage-dependent Na+ channels, and we studied the capacity of NeoSTX administered into the ovary to block sympathetic nerves and PCO phenotype that is induced by estradiol valerate (EV). The toxin was administered with a minipump inserted into the bursal cavity using two protocols: (1) the same day as EV administration and (2) 30 days after EV to block the final step of cyst development and maintenance of the condition. We studied the estrous cycling activity, follicular morphology, steroid plasma levels, and norepinephrine concentration. NeoSTX administered together with EV decreased NA intraovarian levels that were induced by EV, increased the number of corpora lutea, decreased the number of follicular cyst found after EV administration, and decreased the previously increased testosterone plasma levels induced by the PCO phenotype. Estrous cycling activity also recovered. NeoSTX applied after 30 days of EV administration showed near recovery of ovary function, suggesting that there is a specific window in which follicular development could be protected from cystic development. In addition, plasma testosterone levels decreased while those of progesterone increased. Our data strongly suggest that chronic inhibition of sympathetic nerves by a locally applied long-lasting toxin is a new tool to manage the polycystic phenotype in the rat and could be applied to other mammals depending on sympathetic nerve activity.

Steroid Hormones in Reproduction and Roles of GnRH-a in Gonadal Maturation of Marine Fish: A Review

Reproduction in teleosts is regulated by a series of hormones including gonadotropin-releasing hormones (GnRHs), gonadotropins (GTHs) and steroid hormones. To contribute better understanding of steroid hormones in reproduction and GnRH-a in gonadal maturation, this literature review is concerned with the changes of steroid hormone levels in relation with sex inversion, reproductive behavior and gonadal development as well as the application of GnRH-a for inducing maturation of marine fish. The results revealed that in many species of teleost, steroid hormones E2, 11-11-KT and DHP are abundantly produced in gonadal tissues under the control of pituitary gonadotropins, and are essential for critical steps of gametogenesis. Plasma steroid levels have been used as indicators for both of the sex of the fish and its stage in the seasonal reproductive cycle, particularly with regard to induction of spawning. Determination of plasma steroid levels in relation with the sexual status of the gonads over several reproductive seasons might provide valuable information on the mechanisms of sex inversion in ambisexual fish species. In addition, changes of plasma steroid levels in correlation with gonadal development, number of spawning, fecundity, were described clearly in many marine species. The review also indicated that exogenous administration of GnRH-a triggered for final maturation of brood stock of some teleosts. In summary E2, T, 11-KT and C21 steroids are in relation with sex inversion, reproductive processes and GnRH-a is successful for inducing gonadal maturation in some fish species.

Plasma Estradiol and Testosterone Levels and Ischemic Stroke in Postmenopausal Women.

Background and Purpose- Although exogenous hormone therapy (HT) use has been associated with increased risk of ischemic stroke in postmenopausal women, it remains unknown whether sex hormone levels contribute to ischemic stroke risk. We aimed to estimate associations between plasma sex hormone levels and ischemic stroke risk, by HT status, in a nested case-control study of postmenopausal women from the NHS (Nurses' Health Study). Methods- Women with confirmed incident ischemic stroke (n=419) were matched with controls (n=419) by age, HT use, and other factors. Plasma estradiol and testosterone levels were measured using liquid chromatography tandem mass spectrometry; SHBG (sex hormone-binding globulin) was assayed by electrochemiluminescence immunoassay. Associations of total and free estradiol and testosterone, the estradiol/testosterone ratio, and SHBG with ischemic stroke were estimated using conditional logistic regressions stratified by HT status with adjustment for matching and cardiovascular risk factors. Results- Current HT users had different hormone profiles from never/past users. No clear linear trends were observed between estradiol (total or free) levels or the estradiol/testosterone ratio and ischemic stroke risk among either current users (Ptrend>0.1) or never/past users (Ptrend>0.6). For both current and never/past users, the associations between some of the sex hormones and ischemic stroke differed by body mass index categories (Pinteraction≤0.04). For women with a body mass index <25 kg/m2, a higher estradiol/testosterone ratio was associated with significantly elevated ischemic stroke risk among current users (Ptrend=0.01), and higher levels of total and free estradiol were significantly associated with higher ischemic stroke risk among never/past users (Ptrend≤0.04). Testosterone and SHBG were not associated with ischemic stroke in either current or never/past users. Conclusions- Our findings do not support a role of sex hormone levels in mediating ischemic stroke risk among postmenopausal women. Replications in additional larger studies are required.