Journal of Steroid Biochemistry and Molecular Biology最新文献

This study aimed to identify differentially expressed genes (DEGs) in the testes of Bactrian camels during estrus and anestrus and investigate the regulatory role of cholecystokinin (CCK) and its receptor (CCKBR) in androgen synthesis. RNA sequencing was performed on six testicular samples (estrus, n = 3; anestrus, n = 3). A total of 291 DEGs were identified, including 27 upregulated and 264 downregulated in estrus. Gene Ontology (GO) enrichment analysis revealed that CCKBR was significantly enriched in reproduction-related pathways, and STRING analysis showed a close association between CCK and CCKBR. Further qRT-PCR, Western blot, and immunofluorescence (IF) analyses demonstrated significantly higher mRNA and protein levels of CCK/CCKBR in estrus testes (P < 0.01), with both localized in Sertoli cells, Leydig cells, primary spermatocytes, and spermatogonia. Primary Sertoli cells, confirmed by WT1 co-localization, were transfected with p-IRES2-EGFP-CCK and siRNA-CCK. Results showed that CCK overexpression significantly reduced testosterone (T) and dihydrotestosterone (DHT) levels (P < 0.05), while upregulating androgen receptor (AR) and key androgen synthesis enzymes (StAR, P450scc, 3β-HSD) (P < 0.05 or P < 0.01). In contrast, siRNA-CCK exerted the opposite effects. In conclusion, our study highlights the CCK/CCKBR axis as a crucial regulator of seasonal testicular function in Bactrian camels, with CCK negatively regulating testicular androgen synthesis by modulating AR and androgen synthesis enzymes. These findings provide valuable insights into the reproductive biology of Bactrian camels and offer a novel pathway for understanding seasonal fertility regulation in male mammals. This has important implications for enhancing camel breeding efficiency and supporting conservation efforts.

Coffee is widely consumed in Saudi Arabia, but its relationship with vitamin D status and related health indicators remains unclear. This cross-sectional study examined associations between coffee consumption, serum 25-hydroxyvitamin D [25(OH)D], lifestyle factors, and mental health symptoms in 387 adults aged 20-60 years recruited in Saudi Arabia (February-March 2024). Participants were classified as normal (≤3 cups/day) or high (>3 cups/day) coffee consumers. Anthropometric measures and serum 25(OH)D and parathyroid hormone were obtained from medical records, and diet, physical activity, sun exposure, and mental health symptoms were assessed by questionnaire. Associations were examined using group comparisons and multivariable regression models. Compared with normal coffee consumers, high coffee consumers had higher BMI (p = 0.043) and lower serum 25(OH)D (p = 0.05). In multivariable linear regression, higher caffeine intake was associated with lower serum 25(OH)D (β = -0.04 nmol/L per mg/day; 95% CI -0.055 to -0.027; p < 0.001). In logistic regression, higher caffeine intake was associated with lower odds of vitamin D deficiency (<30 nmol/L) (OR = 0.98 per mg/day; 95% CI 0.97-0.99; p < 0.001). High coffee consumers more frequently reported sleep disturbance/insomnia (49.1% vs 34.2%), sweating (20.8% vs 9.6%), and raised heart rate (27.7% vs 17.2%) (all p < 0.01), whereas headache, irritability, anxiety, and depression did not differ between groups. In this sample of Saudi adults, higher coffee intake was associated with lower 25(OH)D, higher BMI, and more arousal-related symptoms. These observational findings warrant confirmation in longitudinal or interventional studies to clarify temporality and inform public health strategies.

Bisphenol A (BPA) has been restricted for its use due to endocrine-disrupting effects and its benzene-ring-substituted derivatives (BPADs) are increasingly used. However, their effects on 11β-hydroxysteroid dehydrogenase type 2 (11β-HSD2), a pivotal enzyme in glucocorticoid regulation, remain poorly characterized. Here, we evaluated, for the first time, how BPADs modulate 11β-HSD2 inhibition, providing mechanistic insights into their endocrine-disrupting potential. This study evaluated six BPADs, identifying 4-hydroxyphenyl-naphthalene (BPH) as the most potent inhibitor (human IC₅₀ = 1.26µM; rat IC₅₀ = 4.17µM), with structure-activity relationships (SAR) revealing critical roles for hydrophobicity (LogP) and steric bulk. Enzyme kinetic inhibition analyses demonstrated competitive or mixed-type inhibition of BPADs, and surface plasmon resonance (SPR) confirmed direct binding of BPH to 11β-HSD2. Molecular docking further highlighted key interactions with residues (Asn167, Lys236) in the steroid-binding pocket. Cellular assays in BeWo cells confirmed functional inhibition of endogenous 11β-HSD2. Notably, human 11β-HSD2 exhibited greater sensitivity than rat ortholog towards BPADs, attributed to a Ser92→Thr92 substitution of 11β-HSD2 sequences. These findings positioned BPADs as novel chemical probes for 11β-HSD2 studies and warranted investigation into their endocrine-disrupting potential, particularly in prenatal contexts where placental 11β-HSD2 safeguards fetal development.

The clinical relevance of serum 25-hydroxyvitamin D [25(OH)D] concentrations in SARS-CoV-2 infection remains uncertain. Most thresholds used to define adequacy were developed for skeletal outcomes, and it is unclear whether they reflect immunerelated risk. To examine whether serum 25(OH)D levels are associated with disease severity, inflammatory and biochemical markers, or clinical outcomes, we conducted a retrospective cohort analysis including 1,136 hospitalized patients with PCR-confirmed SARS-CoV-2 infection. Clinical severity at admission, oxygen requirement, intensive care unit transfer, invasive ventilation, inflammatory markers, length of hospital stay, and in-hospital mortality were compared across vitamin D groups. No significant associations were observed between serum 25(OH)D levels and clinical or laboratory findings. Correlation analyses likewise did not reveal meaningful relationships with inflammatory parameters. In this large hospitalized cohort with predominantly low serum 25(OH)D concentrations, we found no evidence that vitamin D status influenced COVID-19 severity or outcomes. These results indicate that commonly applied adequacy thresholds, including lower immune-related levels, may not provide useful risk stratification in hospitalized patients with COVID-19.

Expression of the critical pyroptosis protein gasdermin E (GSDME) has been reported to be regulated by DNA methylation and negatively correlated with the expression of estrogen receptor (ER) in breast cancer tissues, suggesting that estrogen-induced target gene methylation may be involved in the regulation of GSDME expression in breast cancer cells. To test this hypothesis, we treated MCF-7 and T47D ER-positive breast cancer cells with 17-β-Estradiol (E₂), either alone or in combination with selective ERα antagonist AZD9496, selective ERβ antagonist PHTPP, DNA methyltransferase (DNMT) inhibitor RG108, and selective ER degrader Fulvestrant (Ful). Then, GSDME protein and mRNA expression were examined with western blot and RT-qPCR. Pyroptosis was induced by short-wave ultraviolet (UV-C) and detected with morphological observation, lactate dehydrogenase (LDH) release assay, and propidium iodide-Annexin V-FITC fluorescence staining. The methylation status of the GSDME promoter was tested with methylation-specific PCR. The results demonstrated that 100 nM E₂ significantly decreased GSDME protein and mRNA expression in MCF-7 and T47D cells, and significantly inhibited UV-C-induced pyroptosis. AZD9496 but not PHTPP significantly attenuated the down-regulatory effect of E₂ on GSDME expression. E₂ induced DNA methylation in the GSDME promoter region and up-regulated DNMT1 expression. RG108 strengthened UV-C-induced pyroptosis, and Ful reversed the inhibitory effects of E₂ on UV-C-induced pyroptosis of MCF-7 and T47D cells_. Taken together, our study suggests that E₂ down-regulated GSDME expression in ERα-positive breast cancer by promoting GSDME promoter methylation, and inhibited UV-C-induced pyroptosis.

Obesity, which is mostly related to high fat diet (HFD) consumption, could impair male reproductive function. How vitamin D (VD) co-administration ameliorates male reproductive dysfunction during obesity development is not fully understood. Therefore, this study aims to investigate the effect of VD co-administered with HFD on testicular and sperm functions in mice and to unravel the underlying mechanisms involved.

Methods: Adult male ICR mice were given HFD with VD (HFDVD+) or without VD (HFDVD-), orally for 12 consecutive weeks. Immediately following sacrifice, blood was withdrawn and testes and epididymal sperm were harvested.

Results: HFDVD+ mice exhibited higher serum testosterone, FSH, and LH levels, higher expression of testicular VD receptor (VDR), retinoic acid receptor (RXR α/β/γ) and VD metabolizing enzymes (CYP27B1), testicular steroidogenic proteins (StAR, CYP11A1, 3β-HSD, 17β-HSD, CYP17A1, SF-1 except testicular aromatase), testicular spermatogenic proteins (PLZF, SOX9, SIRT1, AR, SMAD5, ER-α) and blood-testis barrier proteins (occludin, ZO-2, vimentin, connexin-43, N-cadherin) when compared to HFDVD- mice. Additionally, upregulation of testicular RANK and RANKL proteins and downregulation of testicular OPG protein were ameliorated in HFDVD+ mice. Epididymal sperm analysis revealed improvement in sperm parameters in HFDVD+ mice which positively correlated with serum VD levels. In HFDVD+ mice sperm, lesser downregulation of VDR, mitochondrial proteins (TOMM20, ATPB, COX IV), junctional adhesion molecule-A (JAM-A), and glucose transporter 1 (GLUT1) expression were observed.

Conclusion: Co-administration of VD with HFD helps to ameliorate testicular and sperm dysfunctions during obesity development, suggesting VD role in overcoming male reproductive impairment in obesity.