Estradiol Research Articles

Does Vitamin D Administration Increase the Neuroprotective Effect of Estrogen in Male Rats with Traumatic Brain Injury?

Background: In our previous studies, the effect of sex hormones on brain edema reduction after traumatic brain injury (TBI) was demonstrated. In the current study, alone and combined effects of 17-β estradiol (E2) and vitamin D (Vit D) on TBI in male rats were investigated.Methods: Male rats were divided into six groups, including sham, TBI, vehicle, E2, Vit D, and E2+Vit D. In all groups except sham, moderate-intensity diffuse TBI was induced by the Marmarou’s method. Vehicle, E2, Vit D and their combination were intramusculary injected one and 12 hours after the TBI. The brain water content, permeability of blood brain barrier (BBB) and histopathological outcome were assessed 24h after TBI. The neurological outcome score was determined using the veterinary coma scale (VCS).Results: Significant reductions in brain water content (P<0.001, P<0.05 and P<0.01, respectively) and BBB permeability (P<0.001) appeared in the treated groups with E2, Vit D, and E2+Vit D compared to the vehicle group. Twenty-four hours after the injury, the neurological scores in the E2, Vit D, and E2+Vit D groups increased significantly compared to the vehicle group (P<0.05). Dramatic improvement in histopathological outcome was also observed in the treated groups compared to the vehicle group.Conclusion: Alone and combined consumption of estrogen and vitamin D may similarly decrease the development of brain edema and improve the neurological and histopathological consequences of TBI. Therefore, consumption of vitamin D did not enhance the neuroprotective effect of estrogen in TBI.

Isolation of Isoflavones from Iraqi Trifolium pretense

Objective: Trifolium pretense considered one of the most important medicinal plant which belongs to Fabaceae family, this plant commonly known as Red clover, its native region is Europe, Western Asia, and northwest Africa, but planted and naturalised in many other regions. The main compoundes found in Trifolium pretense are isoflavones which is determinrd by genistein, daidzein, biochanin A and formononetin. These compounds structurally related to estradiol-17 beta. Mangement of cancer is the main medicinal uses of Trifolium pretense in addition to decrease cholesterol and LDL and had anti- inflammatory activity, antioxidant and its effect on the Skin, Appendages, and Mucosal Status in Postmenopausal Women. This study was designed to isolate the main isoflavones from Trifolium pretense. Methods: The plant was collecting and washed by tap water to remove dirt and dust and rinsed with distilled water then dried at room temperature. Then, the plant was extracted by ethanol in a soxhlet apparatus and the extract partitioned with ethyl acetate by separatory funnel. The ethyl acetate layer was subjected to standard methods for active constituents identification, and purified by column chromatography. The collected fractions were analysed by TLC comparing with standard isoflavones (genistein, daidzein, biochanin A and formononetin). Preparative TLC was performed using 0.5 mm thickness of silica gel; the isolated bands were conducted with AR grade acetone. Iisolated isoflavones were characterized by 1H and 13-CNMR. Results: Isolated isoflavones (genistein, daidzein, biochanin A and formononetin) were identified by melting points which were identical with to that which were reported in the literature. Moreover, the ¹H-NMR and 13C-NMR analysis were used to identify the target compounds in CDCl3 solvent, and the values of chemical shifts have been discussed according to the literature of analogous compounds.

Transcription factor Sox3 is required for oogenesis in the teleost fish Nile tilapia

Oogenesis is a complex developmental process responsible for the production of eggs from oogonia in fish and other animals. However, transcriptional regulation underlying oogenesis is not fully understood. In the present study, we demonstrated in the teleost fish Nile tilapia that the Sox transcription factor family member Sox3 was involved in regulating oocyte growth during oogenesis. Fluorescence in situ hybridization showed that Sox3 expression was enriched in growing oocytes of ovary but could not be detected in testes. CRISPR/Cas9-mediated homozygous mutation in the Sox3 gene disrupted oocyte growth. Further analysis revealed that Sox3 mutation caused a decrease in the contents of neutral lipids in oocytes and estradiol-17 beta (E2) production. RNA-seq-based transcriptome profiling and RT-qPCR analysis in ovaries demonstrated that the expression levels of genes involved in E2 production, lipid accumulation, and yolk formation were significantly downregulated following Sox3 mutation. Altogether, our findings indicate that Sox3 is required for oocyte growth in Nile tilapia and provides insights into transcriptional regulation underlying oogenesis in teleost fish.

The Effects of BMI and Genetic Variation of Adipokines on Serum Concentrations of Hormones in Untreated Individuals with Breast Cancer; a Pilot Study

Background: Numerous studies have shown an association between hormones secreted by adipose tissues and cancer development. Objectives: This study aimed at investigating the effect of body mass index (BMI) and genetic variation of leptin and adiponectin on serum concentrations of leptin, adiponectin, and estradiol among untreated breast cancer. Methods: This case-control study was performed on 350 women (175 women with breast cancer and 175 healthy controls), who had not taken any medications. Serum levels of estradiol (17-beta estradiol), leptin, and adiponectin were measured, using the ELISA technique. Single-nucleotide polymorphisms of leptin gene (LepG2548A), leptin receptor (Q223, K109R, and K656N), and adiponectin gene (T45G, G276T, C11377G, and 11391A) in blood-isolated DNA were evaluated, using RFLP-PCR technique. Results: Body mass index can affect serum concentrations of hormones and is associated with breast cancer. Also, except for adiponectin C11377G polymorphism, other all genetic variations showed significant relationships with breast cancer. In both groups, BMI was significantly correlated with the mean serum concentrations of hormones, and the risk of breast cancer increased in G2548A, Q223R, K656N, and G276T polymorphisms. The effect of risk allele genotypes on serum concentration of hormones showed that changes in serum concentration of estrogen and leptin in all studied polymorphisms were associated with breast cancer in postmenopausal women. But adiponectin level was only affected by polymorphisms K109R, K656N, and G276 and G11391A. Conclusions: High BMI and genetic variation can affect cancer development by changing the serum concentrations of hormones in different genotypes. Studying various populations’ genetics and lifestyle can help definitive conclusions about genetics and obesity.

Impact of 17-β estradiol on growth and metabolism of marine diatom Thalassiosira weissflogii

• Cultivation of T.weissflogii on synthetically designed media of seawater and estradiol. • Biomass productivity and cell number declined with increase of E2 concentration in samples. • Enhanced production of protein and carbohydrate content mainly in samples T4 and T5. • The level of oxidative stress induced was assessed through MDA and other enzymatic reactions. The inefficiency of existing effluent treatment systems to eliminate pharmaceutical chemicals like endocrine disrupting estrogenic compounds from contaminated waters has raised global environmental and ecological concerns. Diatoms are unicellular, eukaryotic phytoplankton that inhabits ubiquitously and possess adaptive plasticity which enables them to withstand rapid environmental fluctuations. Thus, diatoms can be employed as indicators organisms and bioremediation agents of even trace amounts of Pharmaceutical Contaminants (PCs) like the estrogenic compounds. In this study, the influence of simulated wastewater containing pharmaceutical effluent 17-β Estradiol (E2) on the growth, cellular metabolites and oxidative stress response of the diatom, Thalassiosira weissflogii was assessed. Though an increasing concentration of the chemical displayed a negative correlation with growth, number of cells and biomass productivity; the photosynthetic profile of the diatom showed varied responses with significant enhancement in chlorophyll a and carotenoid content. The test concentrations T1 and T3 showed elevated carbohydrate and protein levels (0.82 ± 0.003 g L -1 and 2.04 ± 0.020 g L -1 ) establishing that E2 stress can be optimised to enhance primary metabolite secretion in diatoms, thereby valorizing its potential as suitable aqua feed. Besides this, an elevation in the antioxidative response against the reactive oxygen species generated was also distinctly recorded with respect to the increased release of catalase (152.70 ± 0.05–265.52 ± 0.06 k mg -1 ) and ascorbate peroxidases (0.959 ± 0.008–1.92 ± 0.021 U mg -1 ) as well as a conspicuous increment in the lipid peroxidation and hydrogen peroxide content was also there. Thus, this novel strategy of employing diatom for multitude of biorefinery aspects besides reacclimating contaminated water reveals the perspective application of this approach. .

A Review on 17-β estradiol a Potent Therapeutic Factor of Diabetic Cardiomyopathy

Type 2 diabetes causes structural and functional changes in the myocardium, which is called cardiomyopathy. Diabetic cardiomyopathy (DCM) is a distinct primary disorder process, independent of coronary artery disease, which leads to heart failure in diabetic patients. Also, DCM is a multifaceted disorder that is one of the leading causes of death in elderly and postmenopausal women. Menopause is associated with decreased and stopped ovarian function, which reduces and stops the production of ovarian hormones, especially estrogen. Moreover, menopause is associated with an increased risk of cardiovascular diseases. Sex steroids such as 17-β estradiol have a variety of protective effects on many tissues in the body, including the cardiovascular system. In this article, the concept of DCM, the underlying molecular signaling pathway, and, finally, the role of 17-β estradiol as one of the most important estrogens in moderating DCM are discussed to provide a theoretical basis for in-depth study.

Exploring the effect of estrogen on Candida albicans hyphal cell wall glycans and ergosterol synthesis.

Increased levels of 17-β estradiol (E2) due to pregnancy in young women or to hormonal replacement therapy in postmenopausal women have long been associated with an increased risk of yeast infections. Nevertheless, the effect underlying the role of E2 in Candida albicans infections is not well understood. To address this issue, functional, transcriptomic, and metabolomic analyses were performed on C. albicans cells subjected to temperature and serum induction in the presence or absence of E2. Increased filament formation was observed in E2 treated cells. Surprisingly, cells treated with a combination of E2 and serum showed decreased filament formation. Furthermore, the transcriptomic analysis revealed that serum and E2 treatment is associated with downregulated expression of genes involved in filamentation, including HWP1, ECE1, IHD1, MEP1, SOD5, and ALS3, in comparison with cells treated with serum or estrogen alone. Moreover, glucose transporter genes HGT20 and GCV2 were downregulated in cells receiving both serum and E2. Functional pathway enrichment analysis of the differentially expressed genes (DEGs) suggested major involvement of E2 signaling in several metabolic pathways and the biosynthesis of secondary metabolites. The metabolomic analysis determined differential secretion of 36 metabolites based on the different treatments’ conditions, including structural carbohydrates and fatty acids important for hyphal cell wall formation such as arabinonic acid, organicsugar acids, oleic acid, octadecanoic acid, 2-keto-D-gluconic acid, palmitic acid, and steriacstearic acid with an intriguing negative correlation between D-turanose and ergosterol under E2 treatment. In conclusion, these findings suggest that E2 signaling impacts the expression of several genes and the secretion of several metabolites that help regulate C. albicans morphogenesis and virulence.

Amaranthus hybridus (Amaranthaceae) prevents the detrimental effects of cyclophosphamide on ovarian function in Wistar rats: An experimental study.

BackgroundCyclophosphamide (CP) is an anticancer agent, but its chronic administration induces ovarian toxicity.ObjectiveWe evaluated the effects of aqueous extract (AE) and methanol extract (ME) of Amaranthus hybridus (A. hybridus) on CP-induced ovarian toxicity in rats.Materials and Methods40 female Wistar rats (10 wk, 170-200 gr) were distributed into 8 groups (n = 5/each) as follows: 1) healthy control; 2) CP+distilled water (10 ml/kg/d); 3) CP+3%-tween 80 (10 mL/kg/d); 4) CP+clomiphene citrate (2 mg/kg/d); 5, 6) CP+AE of A. hybridus (55 and 110 mg/kg/d); and 7, 8) CP+ME of A. hybridus (55 and 110 mg/kg/d). After 28 days of treatment, estrus cyclicity, ovarian and uterine weights as well as estradiol levels and ovarian histology were determined.ResultsCP induced ovarian toxicity after 28 days of exposure. More specifically, CP disturbed the estrus cycle, decreased ovary and uterus weights (p = 0.04), and the 17-β estradiol level (p = 0.04), and induced severe ovarian damages. Remarkably, A. hybridus significantly increased (p = 0.03) the ovarian weight (AE and ME at all doses) and uterus weight (ME at 110 mg/kg/d), compared with the CP-treated rats. Moreover, the 17-β estradiol level was significantly elevated (p = 0.02) in rats given clomiphene citrate and A. hybridus (AE 110 mg/kg/d; ME 55 mg/kg/d). Finally, the ovaries of rats given plant extracts had many corpus luteum and normal follicles, and no cystic follicles.Conclusionprevented the detrimental effects of CP on ovarian function, which could support its traditional use as a fertility enhancer.

Therapeutic Effects of Silibinin Against Polycystic Ovary Syndrome Induced by Letrozole in Rats via Its Potential Anti-Inflammatory and Anti-Oxidant Activities.

BackgroundCurrent therapies for polycystic ovary syndrome (PCOS) are accompanied by unwanted effects. Silibinin; a flavonolignan has pleiotropic activities and favorable safety profile.PurposeTo investigate the efficacy of silibinin on estrous cyclicity, inflammation, oxidative stress and ovarian morphology in letrozole-induced PCOS in rats.MethodsForty-eight female Wistar albino rats were divided into 2 sets. Rats of the first set (n = 12), assigned as a negative control (NC) received only the vehicle, rats of the second set (n = 36), assigned as PCOS rats, were given letrozole 1mg/Kg orally for 21 days. On day 21, six rats from the first set and six rats from the second set were euthanized for confirmation of PCOS-induction. The remaining animals from the first set assigned as group 1, those in the second set (n = 30) were equally divided into 5 groups and treated daily for 19 days as follows: group 2 (positive control) received only the vehicle, group 3 treated with metformin 300mg/Kg orally, groups 4 and 5 treated respectively with 100 and 200 mg/Kg silibinin intraperitoneally (IP), and group 6 treated with a combination of metformin 300mg/Kg orally and silibinin 100mg/Kg IP. On day 40, blood samples were examined for luteinizing hormone (LH), testosterone (TS) and estradiol (EST) levels, the anti-inflammatory and antioxidant parameters, ovarian and uterine morphology.ResultsSilibinin alone or in combination with metformin was found to be effective in restoring the regularity of estrous cycle by ameliorating the abnormal alterations of LH, TS, EST, tumor necrosis factor (TNF)-α, and oxidative status and by resuming the appearance of corpora lutea and decreasing or even total absence of cystic follicles in the ovaries.ConclusionSilibinin was effective in restoring estrous regularities and alleviating hormonal and histomorphological abnormalities of the ovarian and uterine tissues, this could be due to its anti-androgenic, anti-inflammatory and antioxidant properties.

Comparative oestrogenic effects of Allium sativum and Allium cepa in ovariectomised rats

Oestrogens are steroid hormones that influence the growth, differentiation, and functioning of many target organs, including the male and female eproductive organs. Menopause, an important sign of aging in women is characterized by oestrogen depletion, which is associated with many menopausal problems. Oestrogen supplements that are mostly used are not readily safe and have been reported to cause serious health hazards Phytoestrogens are plant compounds that are essentially correspondent to oestradiol (17-β oestradiol) and can relate with oestrogen receptors to elevate and/or prohibit oestrogenic responses. This study therefore aimed at evaluating the possible therapeutic efficacy of Allium sativum and Allium cepa on oestrogenic activities of adult ovariectomised wistar rats. Adult Wistar rats were ovariectomized, then administered various doses of the extracts for 28days, then sacrificed and tissues harvested for analysis. Results shows Allium cepa and quercetin to increase endometrial thickness, increase neural cells in synaptophysin stained cells of hippocampus and dentate gyrus. A conclusion of Allium cepa possessing similar strogenic properties as estradiol group in these ovariectomized rats was drawn, which can offer some ameliorative effects of estrogen deficiency.

Short chain fatty acid, acetate restores ovarian function in experimentally induced PCOS rat model.

BackgroundPolycystic ovarian syndrome (PCOS) is pathogenically characterized with hyperandrogenism and metabolic alterations, which often result in ovarian changes and infertility in women of reproductive age. Epigenetic changes have been linked to the development of PCOS. However, the involvement of epigenetic regulator, histone deacetylase (HDAC) in PCOS-driven ovarian dysfunction is not clear. Howbeit, the present study hypothesized that acetate, an HDAC inhibitor (HDACi) would protect against ovarian dysfunction in experimentally induced PCOS.Materials and methodsFemale Wistar rats weighing 120–150 g were randomly divided into four groups (n = 6). The groups received vehicle, sodium acetate (200 mg/kg), letrozole (1 mg/kg) and letrozole with acetate by oral gavage respectively. The administrations were done daily for 21 days.ResultsThe rat model of PCOS had increased body weight and ovarian weight, 1-hr postload glucose and plasma insulin, testosterone and LH/FSH ratio as well as reduced insulin sensitivity and plasma 17-β estradiol and sex hormone binding globulin. This model of PCOS in addition showed a significant increase in plasma and ovarian triglyceride, total cholesterol, TNF-α and HDAC, and ovarian malondialdehyde as well as a significant reduction in ovarian glutathione peroxidase/reduced glutathione and NrF2 with the histology of ovarian tissues showing disrupted morphology with significant increase in the number of degenerated follicles compared with control group. These alterations were however attenuated when treated with HDACi, acetate.ConclusionAltogether, the present results suggest that acetate protects ovarian function with evidence of normal growing follicles and enhanced circulating 17-β estradiol by inhibition of HDAC.

Mineralocorticoid receptor blockade attenuates hyperandrogenic metabolic dysregulation in letrozole-induced PCOS rat model.

Polycystic ovarian syndrome (PCOS) is a metabolic syndrome associated with mineralocorticoid receptor (MR) activation, which causes infertility in women of reproductive age. Spironolactone (SPL) is a MR blocker with inconclusive effect in the treatment of PCOS. Therefore, the present study hypothesized that low dose SPL would ameliorate metabolic dysfunction associated with PCOS. Female Wistar rats (8-week-old) were divided into 3 groups namely: Control, SPL, Letrozole (LET)-treated and LET + SPL-treated groups. The control group was given vehicle (distilled water), SPL-treated group received 0.25mg/kg, LET-treated group received 1mg/kg of LET and LET + SPL-treated group received a combination of LET and SPL. The administrations were done by oral gavage for 21days uninterruptedly. Biochemical parameters such as lipid profile, malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), γ-glutamyl transferase (GGT), lactate dehydrogenase (LDH), testosterone, 17-β estradiol and glutathione peroxidase (GPx) were determined with appropriate assay methods. Letrozole-treated group had a significant increase in ovarian weight, plasma and ovarian triglycerides, MDA/TNF-α, GGT/LDH and plasma testosterone while it decreased plasma 17-β estradiol and plasma/ovarian high-density lipoproteins and GPx when compared with control group. In addition, histomorphological changes were observed in LET-treated group compared with control group. Nevertheless, administration of low dose SPL attenuated these perturbations. The present study therefore demonstrates that inhibition of mineralocorticoid receptor by low dose SPL ameliorates hyperandrogenic metabolic dysfunction in a rat model of PCOS. Therefore, low dose SPL is hereby suggested as a promising therapeutic agent in the management of PCOS.

Assessment of selected molecular factors and 17-β estradiol dosage in response to Toxoplasma gondii infection in swine.

Background and Aim:Toxoplasma gondii is a global zoonotic parasite infecting virtually all warm-blooded species, although a species-specific variability is evident referring to symptoms frame. Both the success of T. gondii and the outcome of infection depend on a delicate balance between host cellular pathways and the evasion or modulation strategies elicited by the parasite. The hormonal and molecular mechanisms involved in this delicate host-parasite balance are still unclear, especially when considering intermediate host species other than mouse. This study aimed to assess any correlation between T. gondii infection and selected molecular and hormonal factors involved in responses to infection in susceptible species such as swine. Moreover, blood counts and hematochemical assays (glucose, total cholesterol, and triglycerides dosage) were performed to evaluate the overall health condition of animals.Materials and Methods:Toxoplasmosis was diagnosed by enzyme-linked immunosorbent assay for antibodies determination and real-time polymerase chain reaction (RT-PCR) for T. gondii DNA detection. Target genes coding for key factors of cell responses to T. gondii infection were selected, and their transcription was assessed in various tissues by quantitative RT-PCR. 17-β estradiol concentrations were assessed by fluorimetric enzyme-linked immunoassay and the AIA-360 automated immunoassay analyzer. Blood count and hematochemical analyses were performed by a blood cell counter and a spectrophotometer, respectively.Results:The present research highlighted significant differences among infected and uninfected swine (control group) for both transcription profiles of some of the molecular factors considered and 17-β estradiol concentrations. Referring to the assessed hematological and biochemical parameters, no statistically significant differences were observed in infected swine compared to the control group.Conclusion:Our results contribute to the enrichment of data available about the subject and could be useful for a deeper knowledge of the interaction between this parasite and its hosts. However, more aspects are still unclear, such as the effective response of downstream molecules from the same pathways to the variation of factors observed in this study either assessing how the same factors respond to Toxoplasma gondii infection in other host speciesand further analyses should be performed on other host species.

Estrogen May Enhance Toll-Like Receptor 4-Induced Inflammatory Pathways in People With HIV: Implications for Transgender Women on Hormone Therapy.

BackgroundTransgender women (TW) are at increased risk for both human immunodeficiency virus (HIV) and cardiovascular disease (CVD). Antiretroviral therapy-treated HIV has been associated with a two-fold increased risk of CVD, potentially due to dysregulated Toll-like receptor (TLR)-induced immune activation. Use of estrogens in feminizing hormone therapy (FHT) may enhance inflammatory responses and the risk of cardiovascular mortality in TW. Despite this, the immunomodulatory effects of estrogen use in TW with HIV have been inadequately explored.MethodsAs an in vitro model for FHT, cryopreserved PBMCs (cryoPBMCs) from HIV negative (HIV-), HIV+ ART-suppressed (HIV+SP), and HIV+ ART-unsuppressed (HIV+USP) cisgender men were cultured overnight in the presence of 17-β estradiol or 17-α ethinylestradiol with and without the TLR4 agonist LPS or the TLR8 agonist ssPolyU. Monocyte activation (CD69, HLA-DR, CD38) was assessed by flow cytometry. Cytokine levels (IL-6, TNF-α, IL-1β, and IL-10) were measured in cell culture supernatants by Legendplex. Levels of phosphorylated TLR signaling molecules (JNK, MAPK p38) were assessed by Phosflow. Plasma levels of immune activation biomarkers (LPS-binding protein, monocyte activation markers sCD14 and sCD163, and inflammatory molecules IL-6 and TNF-α receptor I) were measured by ELISA.ResultsPBMCs from people with HIV (PWH) produced greater levels of inflammatory cytokines following exposure to LPS or ssPolyU compared to levels from cells of HIV- individuals. While estrogen exposure alone induced mild changes in immune activation, LPS-induced TLR4 activation was elevated with estrogen in cisgender men (CM) with HIV, increasing monocyte activation and inflammatory cytokine production (IL-6, TNF-α). Interestingly, testosterone inhibited LPS-induced cytokine production in CM regardless of HIV status. Plasma markers of immune activation and microbial translocation (e.g., sCD14, sCD163, LPS-binding protein) were generally higher in PWH compared to HIV- CM, and these markers were positively associated with in vitro responsiveness to estrogen and LPS in CM with HIV.ConclusionsOur in vitro data suggest that estrogen exposure may enhance innate immune activation in PWH. Further examination is needed to fully understand the complex interactions of FHT, HIV, and CVD in TW, and determine optimal FHT regimens or supplementary treatments aimed at reducing excess immune activation.

Regulation of Tight Junctions by Sex Hormones in Goat Mammary Epithelial Cells.

Simple SummaryHow ovarian hormones affect goat lactation by regulating cell–cell junctions is still unclear. Through the in vivo and in vitro assays, we found that ovarian hormones could elevate cell–cell junction protein expression, which may affect the intercellular space and molecule transportation between the goat mammary epithelial cells. Our assessment suggests that ovarian hormones may affect goat milk production by regulating the cell–cell junction protein expression between mammary epithelial cells.The sex hormones of estrogen and progesterone (P4) play a vital role in mammary gland development and milk lactation in ruminants. The tight junction (TJ) between adjacent secretory epithelial cells is instrumental in establishing the mammary blood–milk barrier. However, whether estrogen and P4 exert their effect on mammary function via regulating TJ remain unclear. Here, to clarify the role of 17-β estradiol (E2) and P4 in the regulation of TJ in goat mammary gland, we first explored the relationships between the concentrations of E2, P4, and the protein expression of claudin-1, claudin-3, occludin, and ZO-1 during the mammary gland development in goat. Then, we further explored the mRNA and protein expression of claudin-1, claudin-3, occludin, and ZO-1 in the goat mammary epithelial cells (GMECs) in vitro under different concentrations of E2 and P4. The results demonstrated that the protein expression of claudin-1 decreased, but occludin and ZO-1 increased with the decline in E2 and P4 during the transition from pregnancy to lactation. In the in vitro studies, E2 exerted a positive effect on the mRNA expression of claudin-1, and accelerated the proteins’ expression of claudin-1 and ZO-1 in GMECs; P4 upregulated the mRNA expression of claudin-1, claudin-3, occludin, and ZO-1, and also improved the protein expression of claudin-1, claudin-3, and ZO-1 in the GMECs. The results demonstrated that E2 and P4 play an important role in regulating the expression of the mammary TJ components, which may ultimately affect the mammary gland development and milk lactation.

Bindarit Reduces Bone Loss in Ovariectomized Mice by Inhibiting CCL2 and CCL7 Expression via the NF-κB Signaling Pathway.

ObjectiveTo investigate the changes in proinflammatory cytokines and chemokines, namely, C‐C motif ligand (CCL) 2 and CCL7, in postmenopausal osteoporosis (PMOP) and to develop a new drug, bindarit (Bnd), for PMOP in an ovariectomized (OVX) mouse model.MethodsBone marrow macrophages (BMMs) from the femurs of five women with PMOP and five premenopausal women without osteoporosis were detected by RNA sequencing. BMMs from mice were differentiated into osteoclasts and treated with a synthetic inhibitor of CCL2 and CCL7, Bnd, or 17 beta estradiol (E2). Mouse BMMs were differentiated into osteoclasts with or without Bnd for 7 days and analyzed by RNA sequencing. Osteoblasts of mice were induced to undergo osteoblastogenesis and treated with Bnd. OVX mice were treated with E2 or Bnd after surgery. The protein and mRNA expression of CCL2 and CCL7 was detected using immunostaining and qPCR, respectively, in OVX and aged mice and in cells cultured in vitro. Osteoclast formation was detected using a tartrate‐resistant acid phosphatase (TRAP) assay in vitro and in vivo. Alkaline phosphatase (ALP), runt‐related transcription factor 2 (Runx2) and osteocalcin (OCN) were detected using immunostaining to evaluate osteogenesis. Microcomputed tomography was conducted to analyze trabecular bone parameters, the structure model index, bone mineral density and other variables. Nuclear factor‐κB (NF‐κB) signaling pathway‐related protein phosphorylation of IKKα/β (p‐IKKα/β) and p‐NFκB p65 was examined using western blotting.ResultsCCL2, CCL7 and their receptor of C‐C chemokine receptor‐2 (CCR2), and the NF‐κB signaling pathway, were significantly increased in women with PMOP. CCL2 and CCL7 protein and mRNA expression was increased in OVX mice and aged female mice, but the increases were attenuated by E2 and Bnd. E2 and Bnd effectively inhibited osteoclastogenesis and the protein expression of CCL2 and CCL7 both in vitro and in vivo and reduced bone loss in OVX mice. Bnd did not affect the mineralization of osteoblasts directly in vitro but reduced bone turnover in vivo. p‐IKKα/β and p‐NFκB p65 levels were increased in BMMs of mice after differentiation into osteoclasts but were significantly decreased by Bnd.ConclusionThe proinflammatory cytokines and chemokines CCL2, CCL7 and CCR2 were correlated with PMOP. Bnd attenuated the increases in CCL2 and CCL7 levels to affect osteoporosis in OVX mice via the NFκB signaling pathway. Thus, Bnd may be useful as a new therapeutic for the prevention of PMOP.

Suppression of PCSK9/NF-kB-dependent pathways by acetate ameliorates cardiac inflammation in a rat model of polycystic ovarian syndrome

AimEndocrinometabolic disorders in women of reproductive age, including polycystic ovarian syndrome (PCOS) has contributed to increased prevalence of cardiovascular disease (CVD) risk and its attendant complications. Acetate, the most abundant endogenously produced short chain fatty acid has been linked to metabolic health. However, the impact of acetate on CVD-driven pathologies in PCOS is unknown. The present study therefore investigated the effects of acetate on cardiometabolic abnormalities associated with PCOS in rat model, and the possible involvement of PCSK9/NF-kB-dependent pathways. Materials and methodsEight-week-old female Wistar rats were allotted into four groups (n = 6) and the groups received vehicle, acetate (200 mg/kg), letrozole (1 mg/kg) and letrozole plus acetate respectively. The administrations were done once daily by oral gavage and lasted for 21 days. Key findingsIn letrozole-induced PCOS rats characterized with insulin resistance, glucose dysregulation, elevated plasma testosterone and decreased 17-β estradiol as well as degenerated ovarian follicles, there was a significant increase in plasma and cardiac lipid/lipoproteins, lipid peroxidation, inflammatory mediators (NF-kB and TNF-α), γ-glutamyl transferase/lactate dehydrogenase and lactate content, PCSK9 and reduction in plasma and cardiac antioxidants (glutathione peroxidase and reduced glutathione) and plasma nitric oxide synthesis (eNOS and NO) compared with the control rats. In addition, immunohistochemical assessment of cardiac tissue showed severe expression of inflammasome in letrozole-induced PCOS rats compared with the control rats. Nevertheless, supplementation with acetate significantly attenuated these alterations. SignificanceThe present results suggest that acetate protects against cardiac inflammation in a rat model of PCOS by suppression of PCSK9 and NF-kB-dependent mechanisms.

Cyclic estrogen and progesterone during instrumental acquisition contributes to habit formation in female rats

Habit formation is thought to involve two parallel processes that are mediated by distinct neural substates: one that suppresses goal-directed behavior, and one that facilitates stimulus-response (S-R) learning, which underscores habitual behavior. In previous studies we showed that habitual responding emerges early during instrumental training in gonadally-intact female, compared to male, rats. The present study aimed to determine the role of ovarian hormones during instrumental acquisition in the transition from goal-directed to habitual behavior in female rats. Ovariectomized (OVX) female rats were given subcutaneous silastic capsules that released low levels of 17-β estradiol (E2) to maintain estrogen receptor availability. Rats were assigned to one of three hormone treatment conditions: no additional hormone replacement (Control group), replacement with high E2 (High E2 group), or replacement with high E2 followed by progesterone (High E2 + P4 group). Hormone replacement occurred twice during acquisition to mimic natural hormone fluctuations. At test, the Control and High E2 groups demonstrated responding that was sensitive to devaluation by lithium chloride-induced illness, indicating goal-directed behavior. In contrast, the High E2 + P4 group exhibited a pattern of devaluation-insensitive, habitual responding, that suggested the suppression of goal-directed processes. In a follow-up experiment, similar procedures were conducted, however during acquisition, OVX rats were given cyclic high E2 plus medroxy-progesterone (MPA), a form of progesterone that does not metabolize to neuroactive metabolites. In this group, goal-directed behavior was observed. These data indicate that habit formation is not facilitated in low estrogen states, nor in the presence of cyclic high E2. However, cyclic high E2, together with progesterone during acquisition, appears to facilitate the early emergence of habitual responding. Furthermore, these data suggest that a neuroactive progesterone metabolite, like allopregnanolone, in combination with high cyclic E2, supports this phenomenon.

Effects of Abelmoschus manihot Flower Extract on Enhancing Sexual Arousal and Reproductive Performance in Zebrafish.

The flower of Abelmoschus manihot L. is mainly used for the treatment of chronic kidney diseases, and has been reported to have bioactivities such as antioxidant, anti-inflammatory, antiviral, and antidepressant activities. This study used wild-type adult zebrafish as an animal model to elucidate the potential bioactivity of A. manihot flower ethanol extract (AME) in enhancing their sexual and reproductive functions. Zebrafish were fed AME twice a day at doses of 0.2%, 1%, and 10% for 28 days, and were then given the normal feed for an additional 14 days. The hormone 17-β estradiol was used as the positive control. Sexual behavioral parameters such as the number of times males chased female fish, the production of fertilized eggs, and the hatching rate of the fertilized eggs were recorded at days 0.33, 7, 14, 21, 28, and 42. The expression levels of sex-related genes—including lhcgr, ar, cyp19a1a, and cyp19a1b—were also examined. The results showed that the chasing number, fertilized egg production, and hatching rate were all increased with the increase in the AME treatment dose and treatment time. After feeding with 1% and 10% AME for 28 days, the chasing number in the treated group as compared to the control group increased by 1.52 times and 1.64 times, respectively; the yield of fertilized eggs increased by 1.59 times and 2.31 times, respectively; and the hatching rate increased by 1.26 times and 1.69 times, respectively. All three parameters exhibited strong linear correlations with one another (p < 0.001). The expression of all four genes was also upregulated with increasing AME dose and treatment duration. When feeding with 0.2%, 1%, and 10% AME for 28 days, the four sex-related genes were upregulated at ranges of 1.79–2.08-fold, 2.74–3.73-fold, and 3.30–4.66-fold, respectively. Furthermore, the effect of AME was persistent, as the promotion effect continued after the treatment was stopped for at least two weeks. The present findings suggest that AME can enhance the endocrine system and may improve libido and reproductive performance in zebrafish.

Highly Sensitive Simultaneous Measurement of Steroid Hormone in Human Serum by Liquid Chromatography-electrospray Ionization Tandem Mass Spectrometry

Highly Sensitive Simultaneous Measurement of Steroid Hormone in Human Serum by Liquid Chromatography-electrospray Ionization Tandem Mass Spectrometry