Soluble epoxide hydrolase (sEH) is a promising therapeutic target for inflammation-related disorders; however, current synthetic inhibitors are limited due to pharmacokinetic constraints. Marine steroidal metabolites offer a potential source of natural sEH inhibitors. Therefore, this study aims to investigate the sEH inhibitory activity of compounds isolated from the starfish Patiria pectinifera. Six steroidal compounds (1-6) were isolated and evaluated using enzymatic assays, kinetic analysis, molecular docking, and molecular dynamics simulations. Compounds 1 and 2 demonstrated potent sEH inhibition (IC50 = 16.10 ± 2.43 μM and 4.13 ± 1.24 μM, respectively), while compounds 3-6 were inactive. Kinetic studies showed mixed-type inhibition, with lower Kis than Ki values, indicating preferential binding to the enzyme-substrate complex. Computational analyses revealed that compounds 1 and 2 bind to distinct sites within the C-terminal domain with higher affinity than that of the native substrate, inducing conformational changes consistent with allosteric modulation. These findings indicate compounds 1 and 2 as promising allosteric sEH inhibitors, highlighting the potential of P. pectinifera steroids as anti-inflammatory agents.

Ugi-four component reaction of bile acids, formaldehyde, ethyl isocyanoacetate and 1,4-phenylenediamine produced dimeric steroid-glycylglycine conjugates in low yields. The structure of the new compounds consists of two steroid-dipeptide conjugate units linked by a 1,4-phenylene bridge. Detailed NMR characterization provided sufficient evidence on the obtained structures that were confirmed by single crystal X-ray diffraction. Interestingly X-ray crystallography revealed that the dimeric lithocholic acid-glycylglycine conjugate crystalizes in a conformation having the tertiary and secondary amides in the E and Z rotamers respectively.

The ER-α36 isoform is expressed in both ER-positive and ER-negative breast cancer cell lines. ER-α36 is implicated in promoting the proliferation, invasion, and metastasis of tumor cells. Runt-related transcription factor 2 (RUNX2) is widely recognized as a driver of bone-specific metastasis in breast cancer. The study aimed to examine the association between ER-α36 and RUNX2 in breast cancer and its relationship with cancer development. Immunohistochemical analysis of 25 cases revealed subcellular localization patterns of ER-α36 and RUNX2. ER-α36 showed nuclear positivity in 5 cases (20%), cytoplasmic/membranous positivity in 10 cases (40%), and combined nuclear/cytoplasmic positivity in 4 cases (16%). RUNX2 exhibited nuclear positivity in 5 cases (20%), cytoplasmic/membranous positivity in 9 cases (36%), and combined nuclear/cytoplasmic positivity in 6 cases (24%). Case-by-case analysis revealed that in specimens with nuclear ER-α36 positivity, RUNX2 was predominantly localized in the nucleus or nucleus/cytoplasm. Both proteins were predominantly expressed in grade 2 tumors. In vitro experiments, Knockdown of RUNX2 significantly reduced ER-α36 expression by 3.2-fold in MDA-MB-231 cells and by 8.9-fold in MCF-10A[CE] cells (**p < 0.01 and ***p < 0.001, respectively), whereas ER-α36 knockdown significantly decreased RUNX2 expression by 2.3-fold in MDA-MB-231 cells (*p < 0.05). The results of co-immunoprecipitation showed that the two proteins were significantly associated in MCF-10A[CE] and MDA-MB-231 cells. The luciferase reporter assay revealed that down-regulation of RUNX2 inhibited the binding of ER-α36 to estrogen response elements. These results suggest that RUNX2 interacts with ER-α36 and facilitates its transcriptional activation.

Organophosphate esters (OPEs) may disrupt hormonal balance, but their specific effects in pre- and post-menopausal women have been underexplored. This investigation delves into the menopause-specific associations of OPEs with sex hormones in the National Health and Nutrition Examination Survey, 2013-2016. We included 817 premenopausal and 657 postmenopausal women. Survey-weighted multivariable linear regression, restricted cubic splines (RCS), principal component analysis (PCA), and quantile-based g computation (qgcomp) models were conducted to investigate the associations of OPE metabolites and OPEs mixture with sex hormones in pre- and postmenopausal women. In postmenopausal women, bis(2-chloroethyl) phosphate (BCEP) and Bis(1,3-dichloro-2-propyl) phosphate (BDCPP) were negatively associated with testosterone (T) (BCEP: β = -0.06 [-0.10, -0.01]; BDCPP: β = -0.07 [-0.13, -0.01]) and free androgen index (FAI) (BCEP: β = -0.08 [-0.13, -0.02]; BDCPP: β = -0.06 [-0.12, -0.01]). Diphenyl phosphate (DPHP) was also negatively associated with FAI (β = -0.08 [-0.14, -0.02]). RCS analyses revealed non-linear exposure-response relationships between DPHP and T, between BDCPP and estrogen (E2), and between BDCPP and DPHP and the T/E2 ratio. PCA and qgcomp analyses consistently found negative associations between the OPEs mixture and T and FAI. Conversely, in premenopausal women, BCEP was linearly and positively associated with sex hormone-binding globulin (SHBG) (β = 0.05 [0.01,0.09]), whereas DPHP showed a nonlinear relationship with SHBG, with a curve rising then plateauing. Overall, OPEs, alone or as mixtures, might lower T and FAI levels in postmenopausal women. In contrast, BCEP and DPHP might increase SHBG levels in premenopausal women.

Vitamin D deficiency is linked to cardiovascular disease (CVD), with oxidative stress (OS) playing a key role in the etiology of CVD. We hypothesized that vitamin D deficiency induces cardiac oxidative and endoplasmic reticulum stress (ERS), leading to apoptotic cell death. A rat model of vitamin D deficiency and insufficiency was employed for in vivo studies, while the rat cardiomyoblast cell line (H9C2) was used for in vitro experiments. (OS) parameters were assessed by spectrophotometry, gene and protein expression were determined using qPCR and western blotting. OS markers were elevated while antioxidant enzyme activities were decreased in the left ventricle of vitamin D deficient (VDD) rats compared to control rats. VDD downregulated nuclear factor-erythroid factor 2-related factor 2 (Nrf2) and its downstream proteins in the heart. Since elevated OS leads to ERS, we assessed ERS markers in the heart. Levels of ERS markers were elevated in both the VDD and insufficient rats. Pro-apoptotic and autophagy markers such as BAX, BIM and caspase 8 were upregulated in the heart of VDD rats. Experiments with the H9C2 cell line demonstrated that induction of mild OS using H2O2 upregulates ERS, autophagy and apoptotic markers. VD pretreatment of H9C2 cells led to increase in antioxidant enzyme activities, decrease in ERS and autophagy markers and apoptosis. 4PBA, an ERS inhibitor inhibited apoptosis indicative of a direct role for ERS. The findings suggest that vitamin D protects the heart from oxidative injury and cell death by antioxidant defence mechanism, upregulating Nrf2/HO-1 signaling while alleviating ER stress.

A synthesis of hybrids based on esters, amides of the ursane series, and 28-norursane ureas containing terminal alkyne or azide groups was performed via copper-catalyzed azide-alkyne cycloaddition (CuAAC). Derivatives of modified gallic acid, methyl azidobenzoates, 4-(azidomethyl)-3-methyl-1,2,5-oxadiazole-2-oxide, and dimethyl but-2-indiate were used as cyclization partners. The cytotoxicity of the resulting hybrids was evaluated on a panel of human cancer cell lines (MCF-7, HepG2, HeLa, U87MG, A549). Most of the ester and amide derivatives demonstrated weak antiproliferative activity and low selectivity. At the same time, urea conjugates 14a, 17a, 19, 19a showed pronounced cytotoxicity against MCF-7 and HeLa cells. The most active compound 19a effectively inhibited the growth of MCF-7 (IC50 = 6.50 ± 0.49 μM) and HeLa (IC50 = 7.87 ± 1.86 μM), while maintaining low toxicity for normal fibroblasts. The selectivity profile of 19a (SI = 10-12) significantly exceeds that of the reference drug doxorubicin (SI = 0.8-5.6). Hybrids based on 2-azidoethylurea were more active than propargylurea-derived congeners. The deprotection of OH groups of the polyphenol fragment increased cytotoxicity and selectivity. Molecular docking results indicate that urea hybrids with a 1,2,3-triazole linker may have a higher affinity for the ATP-binding site of Akt1 kinase compared to ester and amide analogs. This study substantiates a strategy for the development of selective antitumor agents based on triterpenoid ureas with polar substituents, which are promising for further preclinical optimization.

Hypothalamic-pituitary-adrenal (HPA) axis dysregulation is a hallmark of major depressive disorder in adults. However, the long-term impact of adolescent depression on adult HPA axis function and adrenal steroidogenesis remains unclear. To address this, we exposed male mice to chronic unpredictable mild stress (CUMS) during postnatal weeks (PW) 4-8 to model adolescent depression. At PW8, CUMS mice exhibited elevated HPA axis activity and enhanced adrenal steroidogenesis, confirming successful model induction. By PW16 (adulthood), HPA axis activity had normalized, whereas adrenal steroidogenesis was markedly suppressed. Transcriptomic analysis followed by validation revealed downregulation of Hedgehog signaling components in the adrenal glands of CUMS mice at PW16. Furthermore, a persistent reduction in H3K4me3 enrichment at the SHH promoter was observed from PW8 to PW16. In vitro, corticosterone treatment reduced SHH promoter H3K4me3 levels and inhibited Hedgehog signaling in adrenocortical cells. Collectively, these findings indicate that adolescent CUMS impairs adult adrenal steroidogenesis via glucocorticoid-mediated epigenetic repression of the SHH/Hedgehog signaling pathway. This study provides novel mechanistic insights into the persistent endocrine consequences of adolescent depression.

Colorectal cancer (CRC) remains a leading cause of cancer-related mortality worldwide, and current treatments are limited by low efficacy, toxicity, and resistance. Among phytosterol compounds, β-sitosterol (BST) was identified in abundance in plants and showed potential anticancer activity, as evidenced by several in vitro and in vivo studies. But the poor solubility of BST limits its efficacy toward anti-cancer therapeutics. Research showed that imidazolium-based ionic liquids (ILs) were found to be very effective for increasing the solubility as well as enhancing the bioavailability of anti-cancer drugs. To develop novel phytosterol-based drug candidates with improved anticancer potential, BST was chemically modified through the conjugation with imidazolium-based ionic liquid moieties to generate a series of BST ionic liquid derivatives (BST-ILs). The synthesized derivatives were characterized, and their physicochemical and pharmacokinetic properties were evaluated through in silico ADMET and density functional theory analyses. Notably, the ionic liquid derivatives displayed improved membrane permeability. The biological activities of BST and its BST-ILs derivatives were assessed in the human colorectal cancer (HCT116) cell line. The results demonstrated a concentration-dependent reduction in cell viability, with the ionic liquid conjugates exhibiting significantly enhanced cytotoxicity compared to the parent compound. Furthermore, the derivatives effectively inhibited cell migration and induced apoptosis, as confirmed by wound healing assay and gene expression analyses, including upregulated expression of apoptosis marker genes Bax and Caspase-3 and downregulation of Ki67 and Cyclin D1. Flow cytometric analysis in this study further demonstrated increased apoptotic cell populations following treatment with the derivatives. In addition, western blot analysis revealed altered pro-Caspase-3 protein expression, further supporting the modulation of apoptosis-associated signaling. Molecular docking and molecular dynamics simulations were performed to investigate interactions with key enzymes of the mevalonate pathway, namely farnesyl diphosphate synthase (FDFT1) and squalene synthase (SQS). These findings highlight BST-ILs conjugates as promising steroid-based candidates with the potential to target sterol biosynthesis pathways in colorectal cancer with significant anti-proliferative activities, highlighting the importance of these BST-ILs for further investigation for anti-cancer drug development.

Progesterone, the first sex hormone ever discovered, has a long history. Yet, it gained prominence in research during the 1930s. Currently, our understanding of progesterone goes beyond its role in the reproductive system, also encompassing the cardiovascular system. Studies have reported on the protective role played by this hormone in the cardiovascular system, which is primarily due to its actions on the vasculature. Progesterone stimulates vasoactive factors, modulates lipid profiles, reduces inflammatory responses, inhibits platelet aggregation and cell proliferation, and lowers blood pressure, thereby contributing to the reduction of cardiovascular disease risk. However, for a long time, hormone replacement therapy relied on the use of progestins, synthetic compounds that failed to replicate the beneficial effects of progesterone and often cause various adverse effects, which cannot be attributed to progesterone itself. In this context, this review compiles the most recent findings regarding the actions of progesterone in the vascular system, while also presenting a historical overview that contextualizes the research in both clinical and experimental areas related to progesterone. Thus, we seek to clarify the unique role played by progesterone in the cardiovascular system, dispelling misconceptions about its effects and discussing its therapeutic prospects.

Canine distemper virus (CDV) induces central nervous system (CNS) demyelination, potentially influenced by local glucocorticoid metabolism via 11β-HSD enzymes and glucocorticoid receptors (GR). This study investigated the relationship between demyelination and local glucocorticoid synthesis in CDV infection. Cerebellar tissues from CDV-infected dogs (diseased group; n = 20) and clinically healthy dogs (control group; n = 10) were analyzed. Sections were immunohistochemically stained with anti-CDV, anti-11β-HSD1, anti-11β-HSD2, and anti-GR antibodies. In the diseased group, 11β-HSD1 immunopositivity was significantly higher than in controls in endothelial cells (p < 0.01), reactive astrocytes (p < 0.001), Purkinje neurons (p < 0.01), and molecular layer neurons (p < 0.001). 11β-HSD2 immunopositivity was significantly increased in reactive astrocytes (p < 0.01), white matter neurons (p < 0.001), and Purkinje neurons (p < 0.01). GR immunopositivity was elevated in reactive astrocytes (p < 0.01) and mossy fibers (p < 0.001). A strong positive correlation was observed between demyelination severity and 11β-HSD1 in reactive astrocytes (r = 0.915, p < 0.01), whereas a moderate negative correlation was found between demyelination severity and GR in reactive astrocytes (r = -0.461, p < 0.05). These findings indicate that local glucocorticoid metabolism is associated with CNS demyelination during CDV infection. This study provides novel insights into 11β-HSD enzymes and GR in CNS pathology, offering a translational model for understanding local glucocorticoid regulation in human demyelinating diseases such as multiple sclerosis.

Ten undescribed hydroxylated, acetylated, esterified and glycoside derivatives of lithocholic acid (LCA) were obtained through the biotransformation of Aspergillus minisclerotigenes, alongside four known compounds. Structural characterization employed integrated NMR spectroscopy, HR-ESI-MS, and X-ray crystallography. Biological evaluation identified compound 10 as a potent inhibitor of LPS-induced NO production in BV2 microglial cells (IC50 = 4.67 μM) and dose-dependently suppressed mRNA expression of IL-1β, TNF-α, iNOS, and COX-2. Furthermore, compound 10 promoted M2-polarization of LPS-activated microglia to attenuate neuroinflammation. Subsequent analysis demonstrated its induction of inflammatory autophagy in BV2 cells. These results suggested that biotransformation was an efficient strategy for structural diversification of LCA and discovery of potent anti-inflammatory leads.