Granulosa cells (GCs) are essential for follicular development, and their apoptosis is a key determinant of follicular atresia in mammals. Circular RNAs (circRNAs) have emerged as critical post-transcriptional regulators in reproductive biology; however, their roles in porcine GC apoptosis remain incompletely characterized. A novel circRNA derived from exon 2 of the INHBA gene (circINHBA) was identified from previous circRNA sequencing data. Its circular structure was verified by RNase R digestion assay, and subcellular localization was assessed by nucleocytoplasmic separation and fluorescence in situ hybridization. The functional role of circINHBA was investigated by siRNA-mediated knockdown, FACS, and Western blotting. The ceRNA mechanism was examined using dual-luciferase reporter assays, RNA pull-down, and RNA immunoprecipitation. circINHBA exhibited significantly higher expression in healthy follicles than in early atretic follicles and was predominantly localized in the cytoplasm. Knockdown of circINHBA promoted GC apoptosis and decreased the BCL2/BAX ratio. Mechanistically, circINHBA functioned as a molecular sponge for miR-196b-5p, which directly targets the 3'UTR of PARP8, thereby upregulating PARP8 expression to inhibit apoptosis. Furthermore, EIF4A3 physically interacted with circINHBA, and its knockdown significantly reduced circINHBA expression, indicating that EIF4A3 promotes circINHBA biogenesis. The EIF4A3/circINHBA/miR-196b-5p/PARP8 axis constitutes a novel regulatory mechanism governing GC apoptosis and follicular atresia in pigs, providing new insights into circRNA-RNA-binding protein crosstalk in ovarian physiology.

In diabetic kidney disease (DKD), overactivation of the renin-angiotensin-aldosterone system (RAAS) promotes ferroptosis. However, the specific role of aldosterone-the terminal effector of the RAAS-in regulating ferroptosis pathways in DKD remains poorly understood. In this study, we investigated how hyperaldosteronism modulates ferroptosis in glomerular mesangial cells. Using db/db mice and high-fat diet (HFD)-induced mice as models, we characterized the metabolic disturbances associated with DKD and the concurrent elevation of endogenous aldosterone. In vitro, MES13 SV40 mesangial cells were exposed to high levels of glucose and aldosterone to recreate the DKD microenvironment, thereby enabling the identification of critical pathogenic targets. We found that db/db and HFD-induced mice exhibited elevated endogenous aldosterone levels along with renal ferroptosis. Aldosterone aggravated ferroptosis in glomerular mesangial cells under high-glucose conditions by increasing ROS, lipid peroxidation products, and malondialdehyde (MDA), while reducing intracellular glutathione (GSH). These changes were coincident with upregulation of glutathione-specific γ-glutamyl cyclotransferase 1 (CHAC1) and downregulation of GPX4 in mesangial cells. Pharmacological inhibition with a mineralocorticoid receptor antagonist or a ferroptosis inhibitor, or genetic knockdown of CHAC1, partly reduced lipid peroxidation, lowered intracellular ROS and MDA levels, and elevated GSH levels and GPX4 expression in mesangial cells. Moreover, treatment with ferroptosis agonists suppressed GPX4 expression, whereas CHAC1 knockdown significantly attenuated this effect. Collectively, these findings indicate that aldosterone, in combination with high glucose, promotes ferroptosis-related phenotypes in mesangial cells, accompanied by CHAC1 upregulation and CHAC1-associated GSH/GPX4 dysregulation, thereby impairing GSH/GPX4-mediated antioxidant defense. These results further implicate CHAC1-associated ferroptosis in DKD pathogenesis and suggest that CHAC1 may serve as a key mediator through which aldosterone exacerbates ferroptosis under hyperglycemic conditions.

Estradiol-17β (E2) regulates many physiological processes in fish. E2 and thyroid hormone (TH) interact in fish, but their effects on the stressed fish brain remain poorly understood. We hypothesised that E2 may interact with TH via their receptors and regulators, thereby affecting energy metabolism during stress acclimation in fish brain segments. We thus examined the short-term (60 min) in vivo effect of E2 on the expression patterns of the E2 receptor-aromatase (ER-arom) system, such as erα, erβ, cyp19a1a, cyp19a1b, and the thyroid hormone receptor and regulator (TR-R) system, such as trα, trβ, trαb, deo1, deo2, deo3, mct8, mct10, as well as energy provision in the brain segments of female air-breathing fish (Anabas testudineus) held under net-confined (NC) stress conditions. NC stress activated all these transcripts across the brain segments, most prominently in the mesencephalon (MC) segment. E2 selectively modulated ER-arom and TR-R transcript expression across brain segments of NC-stressed fish. The Venn diagram analysis showed that E2 enhanced several receptor gene transcripts (erβ, cyp19a1b, trα, trβ, trαb) and attenuated regulatory (Deo) or transport (Mct) genes in MC segments. Furthermore, E2 shifted energy metabolism towards anaerobic pathways in the stressed MC segment, as indicated by a reversal of the increase in COX activity and a decrease in LDH activity. These data indicate that E2 plays a crucial role in coordinating ER-arom and TR-R systems under stress, suggesting that E2 may safeguard the fish brain by differentially regulating these systems and reversing high energy demands across brain segments during stress acclimation.

Clinical evidence suggests a complex link between inflammatory bowel disease and systemic glucose homeostasis, yet the underlying molecular mechanisms remain elusive. Here, we report that chronic ileitis, induced by dextran sulfate sodium (DSS) or IL10 deficiency under a high-fat diet(HFD), paradoxically ameliorates systemic glucose intolerance and preserves pancreatic β-cell mass in mice. Multi-omics analysis of ileal contents revealed a specific enrichment of Akkermansia muciniphila (AKK) and elevated levels of the metabolite 3-hydroxybutyrate (3-HB). Mechanistically, 3-HB activated HCAR2/CREB-associated signaling and increased NKX6.1 expression. NKX6.1 loss-of-function markedly attenuated 3-HB-induced insulin gene expression and glucose-stimulated insulin secretion, establishing NKX6.1 as an important functional mediator of the β-cell response to 3-HB. Furthermore, exogenous administration of 3-HB recapitulated these protective effects in diabetic mice. This study identifies a novel gut-islet axis where microbiota-derived 3-HB preserves β-cell functional identity via HCAR2 dependent regulation of NKX6.1, offering a potential therapeutic strategy for type 2 diabetes.

Impaired insulin secretion, a hallmark of type 2 diabetes (T2D), is associated with the downregulation of syntaxin 1A (encoded by Stx1a), a core SNARE protein essential for insulin exocytosis, in pancreatic β-cells. However, the mechanism regulating syntaxin 1A expression remains unclear. This study aimed to determine the role of the nuclear receptor farnesoid X receptor (FXR) in modulating syntaxin 1A expression. We demonstrated that syntaxin 1A expression was significantly reduced in islets from diabetic Goto-Kakizaki (GK) rats and in INS-1832/13 cells exposed to chronic high glucose. FXR levels were similarly diminished under these diabetic conditions. Genetic knockdown or knockout of FXR in β-cells markedly decreased syntaxin 1A expression and impaired glucose-stimulated insulin secretion (GSIS). Conversely, activation of FXR with GW4064 or chenodeoxycholic acid (CDCA) robustly upregulated syntaxin 1A expression. Chromatin immunoprecipitation assays revealed that FXR activation enhanced its binding to a conserved response element in the Stx1a promoter, facilitating the recruitment of the coactivator steroid receptor coactivator-1 (SRC-1) and increasing histone H3 acetylation, an epigenetic marker for active transcription. Intriguingly, FXR also promoted its own expression through a positive autoregulatory loop. Collectively, these findings identify FXR as a key transcriptional regulator of syntaxin 1A in β-cells. The impairment of the FXR-syntaxin 1A pathway contributes to the secretory dysfunction of diabetic β-cells, revealing a novel pathogenic mechanism underlying T2D.

Thyroid dysgenesis (TD) is the main cause of congenital hypothyroidism (CH). However, a definitive genetic cause is confirmed in only 2-5% of patients, since pathogenic mutations are rare. This study was conducted to advance the understanding of TD genetic basis by investigating genetic alterations in the exomes of affected individuals. A total of 71 TD patients were included; 35 underwent whole-exome sequencing (WES), while the remaining 36 patients and 128 Brazilian non-TD controls were genotyped by Sanger sequencing. Data from the Online Archive of Brazilian Mutations and 1000 Genomes Project were used for comparative analyses. WES revealed a high frequency of FOXE1 polymorphisms and five rare heterozygous DUOX2 variants previously identified in CH patients: p.F966SfsX29 (pathogenic); p.R516H (likely pathogenic), p.T423I and p.N1170T (variant of uncertain significance, VUS) and p.A1123T (likely benign) according to ACMG criteria. Patients with DUOX2 variants also harbored FOXE1 polymorphisms. Regarding population-wide risk, functional FOXE1 rs1867277 allele A and the rs71369530 14Ala allele were associated with TD, individually, and in combination in the Brazilian cohort, not previously reported. Functional analysis of DUOX2 variants showed disruption of epithelial-mesenchymal transition-related gene expression without affecting 3D spheroid formation; only the p.F966SfsX29 variant reduced hydrogen peroxide generation. In silico protein homology-based modeling revealed altered structures of all DUOX2-variants-DUOXA2 complex, including hydrogen bond position changes. In conclusion, our findings confirm two FOXE1 polymorphisms as TD risk factors, show that their combined inheritance increases disease risk, and reveal that rare DUOX2 variants co-occur with FOXE1 risk polymorphisms, supporting a polygenic model.

Thyroid hormone receptor (TR) activation of thyroid hormone (T3)-responsive gene expression is enhanced by receptor acetylation, but paradoxically, we previously showed that TR acetylation mimics have a more cytoplasmic localization compared to nonacetylation mimics. Here, we showed that a substitution mutation at any of the three acetylation sites is sufficient for altered localization and transcriptional activity in transfected human cells. Inhibition of select deacetylases showed no cytoplasmic shift for TR, whereas inhibition of the acetyltransferase CBP/p300 significantly increased TR nuclear localization, suggesting that acetylation, not deacetylation, drives localization. Using acetyl-lysine immunoprecipitation assays, we observed a greater population of acetylated TRα1 in the nucleus compared to the cytoplasm; however, acetylated TRα1 was more susceptible to nuclear proteases, suggesting that shuttling to the cytoplasm is a protective mechanism for modulating the extent of nuclear degradation of acetylated TR. Ubiquitin immunoprecipitation assays revealed significantly elevated polyubiquitination of the acetylation mimic, and biochemical fractionation revealed that polyubiquitinated wild-type TRα1 is enriched in the nuclear matrix, with no detectable modified TRα1 in the cytoplasm. Further, proteasomes shift toward a more nuclear localization in TRα1-expressing cells. Resistance to Thyroid Hormone syndrome α (RTHα) mutant C392X had significantly increased acetylation levels but a wild-type polyubiquitination profile, suggesting that aberrant acetylation is an additional characteristic of the molecular pathology RTHα. Our findings provide novel insights into TR's life cycle supporting a model in which TR exists primarily in the nonacetylated state, while acetylation promotes T3-responsive gene expression, followed by TR polyubiquitination, and rapid T3-dependent degradation by nuclear proteasomes.

Mitochondrial dysfunction has been implicated in various female reproductive disorders, but the underlying molecular links remain poorly understood. Here, we systematically investigated the genetic associations between mitochondrial-related genes and 14 female reproductive disorders through an integrative analysis of multi-omics datasets. Summary-level data on mitochondrial gene expression, DNA methylation, and protein abundance were obtained from multiple quantitative trait loci (QTL) resources and jointly analyzed in relation to 14 common female reproductive disorders. By integrating genetic association signals across molecular layers, we identified multiple mitochondrial-related genes showing nominal candidate associations with reproductive outcomes. Notably, genetically predicted NEU4 expression was nominally associated with lower risks of ovarian dysfunction and polycystic ovary syndrome. At the methylation level, NEU4 was represented by 25 probe-trait associations across six reproductive phenotypes. At the proteomic level, higher genetically predicted NUDT9 abundance was associated with abnormal uterine bleeding. Overall, 12 associations survived false discovery rate (FDR) correction, of which 10 also satisfied the heterogeneity in dependent instruments (HEIDI) threshold. ALDH2 remained a recurrent nominal and pharmacologically annotated candidate. These findings provide an exploratory molecular landscape and prioritize a smaller set of FDR-supported signals for independent replication and mechanistic investigation.

This review examines metabolic comorbidities affecting the heart, liver and kidney. The clinical prevalence of cardiohepatorenal metabolic comorbidities associated with obesity and type 2 diabetes is substantial. Chronic low-grade inflammation serves as the fundamental pathogenic mechanism. Abnormal macrophage activation is associated with metabolic disorders and damage to multiple organs. The conventional M1/M2 binary classification fails to adequately capture the heterogeneity of macrophages in vivo. Previous studies have primarily focused on the investigation of individual organs, leaving the cross-organ immune-metabolic linkage mechanism poorly understood. Furthermore, targeted interventions often lack precision. In this study, we systematically summarize the roles of macrophage polarization, phenotypic remodeling and metabolic activation in the injury of three metabolic organs. We also analyze their mediating functions in inter-organ immune-metabolic crosstalk and the associated pathological connections. Glycolipid toxicity impairs the glycolipid metabolism of macrophages via the NLRP3/SIRT pathway, resulting in a pro-inflammatory polarization advantage. Specific subsets of macrophages, including CCR2[+],TREM2[+],and CD206[+]CD68[+],are involved in mediating myocardial remodeling, liver fibrosis, and renal interstitial injury, respectively.Medications such as SGLT2 inhibitors and GLP-1 receptor agonists may provide multi-organ protection by modulating macrophage activity. The current research presents several limitations, including species differences between humans and mice, ambiguous temporal transformation rules of macrophages, and inadequate specificity of targeted preparations.Future efforts should leverage spatial multi-omics and gene editing technologies to analyze the global regulatory network and develop highly selective targeted strategies. This approach can offer theoretical support for the integrated and precise prevention and management of metabolic comorbidities.

Deoxynivalenol (DON) is a highly prevalent mycotoxin contaminating cereals and cereal-based products and is increasingly recognized for its adverse effects on reproductive function. DON is known to activate mitogen-activated protein kinases (MAPKs), which are also critical mediators of gonadotropin-releasing hormone (GnRH)-dependent synthesis and secretion of luteinizing hormone (LH) and follicle-stimulating hormone (FSH) by pituitary gonadotrope cells. Despite this mechanistic overlap, the direct endocrine-disrupting effects of DON on pituitary gonadotropin production remain poorly understood. Here, we demonstrate that exposure of murine gonadotrope-like LβT2 cells to DON compromises cell viability at nanomolar concentrations and reduces GnRH-induced LH production at higher concentrations in a concentration-dependent manner. Notably, distinct biological endpoints displayed different sensitivity thresholds, with viability being affected at lower concentrations than those required to alter GnRH-dependent signaling and hormone production. Mechanistically, DON promotes an apoptotic phenotype and selectively attenuates GnRH-induced extracellular signal-regulated kinase (ERK) phosphorylation. This attenuation of GnRH-induced ERK activation was associated with reduced expression of Cga and Lhb, two key genes involved in LH synthesis and secretion. Collectively, these findings identify the pituitary gland as a direct target of DON toxicity and provide evidence that DON directly targets pituitary gonadotrope-like cells and can disrupt GnRH-dependent endocrine function under exposure conditions that impair intracellular signaling and hormone production, thereby expanding current understanding of the neuroendocrine mechanisms underlying DON-induced reproductive dysfunction.

Menopause is associated with visceral obesity, low-grade chronic inflammation, and increased risk of dyslipidemia, insulin resistance, and type 2 diabetes. Although exercise (EXE) and GLP-1 receptor agonists, such as semaglutide (SEM), have demonstrated beneficial effects on metabolic regulation, their combined impact in menopause models has been less investigated. In this study, ovariectomized female Wistar rats were divided into four experimental groups: sedentary plus saline (SED + SAL), sedentary plus semaglutide (SED + SEM), exercise (EXE + SAL), and EXE + SEM. The aerobic exercise protocol consisted of treadmill running at moderate intensity for four weeks, during which the rats received either SEM or SAL weekly. At the end of the protocol, metabolic parameters were assessed in blood, liver, soleus and gastrocnemius muscles. Both SEM groups had lower body weight, weight gain, relative subcutaneous white adipose tissue (scWAT) mass, glucose, total proteins, and triglycerides in serum and liver, while LDL cholesterol, glycogen in liver and gastrocnemius, and AMPK and HSF1 expression in gastrocnemius increased. EXE decreased total and LDL cholesterol, liver triglycerides and HSF1 expression in both tissues, while glycogen in liver and gastrocnemius, liver HSF1 expression and relative mass were increased. The combination of SEM + EXE increased glycogen content in the gastrocnemius and AMPK expression in both gastrocnemius and liver. This combined treatment reduced HDL levels, hepatic triglycerides and scWAT relative weight. These results suggest that SEM and EXE exert complementary effects on glycemic and lipid metabolism, with partially distinct metabolic outcomes that may involve different underlying mechanisms.