Parkinson's disease (PD) is a progressive neurodegenerative disorder. An essential early hallmark of PD is disrupted mitochondrial dynamics driven by impaired cellular energy homeostasis. Therapeutic interventions restoring mitochondrial function and biogenesis hold promise for neuroprotection in PD. In the present study, the neuroprotective effects of formononetin (FMN) were evaluated in both MPP+-induced SH-SY5Y cells and the MPTP-induced mouse model of Parkinson's disease, using concentrations of 5, 10, 20, and 40 μM in vitro and doses of 25 and 40 mg/kg in vivo. To evaluate its biological activity, we employed western blotting and immunofluorescence assay to quantify the expression of disease-relevant markers. Mitochondrial health was further assessed using Mitotracker, alongside reactive oxygen species (ROS) assessment. Motor behavior and molecular endpoint parameters were also measured. Our results demonstrated that FMN significantly attenuates MPP+/MPTP-induced neurotoxicity, improves motor function, and restores the expression of PGC-1α, tyrosine hydroxylase, and key mitochondrial proteins involved in mitochondrial DNA replication. Enhancement of mitochondrial fusion proteins and other transcriptional regulators was also observed in the administered groups. Flow cytometry and imaging analyses confirmed that FMN-mediated PGC-1α activation preserves mitochondrial integrity and reduces oxidative stress. Altogether, these findings provide evidence that formononetin exerts neuroprotective effects in PD by modulating the PGC-1α signaling axis.
Exosomes are nanoscale extracellular vesicles (EVs) that have recently garnered significant attention owing to their crucial role in orchestrating cell-to-cell communication. Through the transfer of heterogeneous molecular cargo encompassing lipids, proteins, cytokines, growth factors, and RNAs (including mRNAs, lncRNAs, miRNAs, and circRNAs), they modulate a wide spectrum of physiological and pathological processes. Exosomes have been extensively investigated as diagnostic tools, therapeutic agents, as well as innovative platforms for drug delivery in metabolic, oncological, cardiovascular, and neurological disorders. Culminating evidence has demonstrated the pivotal role of exosomes in renal pathophysiology. Depending on their cargo content, exosomes represent potential biomarkers for early disease detection and survival prediction across various renal pathologies. While current therapeutic interventions are largely confined to attenuating disease progression, exosomes hold the potential to promote regeneration in both acute kidney injury and chronic kidney diseases. The current review comprehensively examines the clinical utility of exosomal cargo as diagnostic and prognostic biomarkers as well as therapeutic agents in kidney diseases, highlighting their crosstalk with critical signaling pathways implicated in renal pathophysiology. Addressing the current challenges in exosome isolation and standardization, and the development of advanced exosome engineering technologies are crucial for the transformation from experimental research settings to clinical practice. This should be augmented by preclinical validation and well-designed clinical trials, ultimately paving the way for a new era of precision medicine.
Osteoporosis is closely linked to oxidative stress and inflammation, positioning the vitamin E metabolite γ-CEHC, known for its robust antioxidant and anti-inflammatory properties, as a promising therapeutic agent. However, its molecular targets have remained largely unknown. In this study, we characterized the protein targets of γ-CEHC and clarified its role in regulating bone metabolism using an ovariectomized (OVX) mouse model and in vitro assays. Bone morphological analysis and histomorphometry demonstrated that γ-CEHC improves osteoporosis in OVX mice by inhibiting osteoclast differentiation and enhancing osteoblast differentiation. To identify the underlying mechanisms, we employed isothermal thermal proteome profiling (TPP) to map γ-CEHC-interacting proteins, followed by Gene Ontology (GO) and KEGG enrichment analyses. Our findings identified fatty acid-binding protein 5 (Fabp5) as a core target. The direct and specific binding between γ-CEHC and Fabp5 was confirmed through cellular thermal shift assays (CETSA), molecular docking-suggesting hydrogen bonding with Thr63-and Surface Plasmon Resonance (SPR) which showed a strong binding affinity (Kd = 5.24 μM). Furthermore, γ-CEHC was found to suppress LPS-induced M1 macrophage activation and promote M2 polarization, thereby reducing reactive oxygen species (ROS) levels and restoring bone remodeling homeostasis. This study is the first to systematically elucidate the molecular mechanisms of γ-CEHC in bone metabolism, revealing that it acts as a highly selective ligand for Fabp5. These findings provide a novel mechanistic basis for using γ-CEHC and targeting Fabp5 in the treatment of osteoporosis.
Fermented foods are increasingly recognized for their health-promoting properties, but little is known about the effects of sauerkraut brine (SB), a by-product of cabbage fermentation, on systemic inflammation and neuro-inflammatory responses. This study aimed to investigate the immunomodulatory, antioxidant, and behavioral effects of SB oral treatment in a mouse model of low-dose lipopolysaccharide (LPS) challenge. The SB was prepared by traditional spontaneous cabbage fermentation and analyzed for pH, microbiological profile, and acidity. At the end of fermentation, the brine contained a high level of viable lactic acid bacteria (LAB) (1.8 × 106 CFU/ml). Mice were pretreated orally with SB, heat-treated SB (htSB), or saline for 4 weeks before single or repeated LPS injection (0.5 mg/kg). Safety assessments, including body weight, food intake, and locomotor activity, did not indicate adverse effects with either form of SB. In the acute LPS model, SB pretreatment significantly reduced mRNA expression of the pro-inflammatory cytokines TNF-α, IL-1β, and IL-6 in the prefrontal cortex compared to the saline-pretreated group. Interestingly, the expression of IL-10, strongly induced by LPS, was also significantly reduced by SB, suggesting modulation of both pro- and anti-inflammatory signaling pathways. These protective effects were less pronounced in the heat-treated SB group, suggesting that viable bacteria or heat-sensitive components may be critical for bioactivity. In the repeated LPS model, SB prevented LPS-induced depletion of glutathione (GSH) and preserved total antioxidant capacity (TAC), while heat-treated SB provided no protection and resulted in increased protein carbonylation and decreased TAC. No significant changes in the activities of antioxidant enzymes (SOD1/2, CAT, GR, GSH-Px, GST) were observed. Behavioral tests using the open-field paradigm showed that all groups exhibited prolonged sickness behavior after repeated LPS exposure, despite a two-day washout period. However, only the SB group showed delayed recovery, with reduced horizontal and vertical activity lasting 1 day longer than in the other groups. Nevertheless, pretreatment with SB and heat-treated SB prevented the LPS-induced reduction in time spent in the central zone, a commonly used measure of anxiety-like behavior, suggesting a possible anxiolytic effect. In conclusion, SB exhibits anti-inflammatory and antioxidant properties associated with systemic LPS-induced neuroinflammation and may reduce anxiety-like behavior. These effects appear to be dependent on heat-sensitive constituents or microbial viability. The findings support further investigation of SB as a functional dietary intervention targeting neuro-immune health.
Glioblastoma (GB) is highly malignant with a median survival of 14 months despite conventional treatments like surgery, radiotherapy, and temozolomide. Resistance to these therapies necessitates innovative approaches, such as immune checkpoint inhibitors (ICIs) targeting cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed cell death protein 1 (PD-1), and programmed death ligand 1 (PD-L1) to enhance T-cell-mediated tumor destruction. However, clinical trials have shown limited ICI efficacy in GB due to its immunosuppressive microenvironment and the blood-brain tumor barrier (BBTB), which impairs drug delivery. Emerging evidence highlights the gut microbiota as a pivotal modulator of ICI response, enhancing CD8+ and CD4+ T-cell function, antigen presentation, and immune modulation via the gut-brain axis in cancers. In addition, studies showed that gut-derived metabolites, including short-chain fatty acids, modulate immune responses and support blood-brain barrier integrity by regulating inflammatory signaling and tight junction proteins. Future GB research should prioritize clinical trials, mechanistic studies, and interventional strategies like fecal microbiota transplantation and probiotics to enhance ICI efficacy.
Herein, we adopted a dual approach combining molecular modeling and biological studies, in order to assess the interaction between four selected natural antioxidants (NAs; berberine, curcumin, astaxanthin, indicaxanthin) and tissue transglutaminase (TG2) levels both in the absence and in the presence of full native peptide of amyloid-β (Aβ). Docking studies were performed to ascertain the binding affinity of NAs against the TG2 closed, Ca2+-bound closed, and open forms. In the biological investigation, the effect of berberine and curcumin treatment on TG2 in Olfactory Ensheathing Cells (OECs) exposed to Aβ(1-42) or to Aβ(25-35), a Aβ toxic fragment, or to reverse-sequence fragment Aβ(35-25), an Aβ not toxic fragment, was tested. In addition, their effect on the percentage of cell viability and cytoskeleton marker (GFAP, vimentin and nestin) levels were evaluated. The role of berberine and curcumin on both endocellular levels of reactive oxygen species (ROS) and apoptotic pathway activation were also assessed. Our findings demonstrate that pretreatment of OECs with these NAs counteracted the Aβ-induced upregulation of TG2, restoring its expression to control levels and preserving its predominant cytosolic localization. Furthermore, antioxidant pretreatment reinstated cell viability, normalized the expression of GFAP, vimentin, and nestin, reduced intracellular ROS accumulation, and prevented activation of the apoptotic cascade. Our findings demonstrate that integrating computational and biological approaches, enhances the identification of potent therapeutic agents and also highlights berberine and curcumin as promising candidates for the development of novel neuroprotective drugs against neurodegenerative disorders, including Alzheimer's disease.
Ovarian response extremes are associated with distinct alterations in the follicular microenvironment, yet the underlying molecular mechanisms remain incompletely understood. This study investigated oxidative stress, antioxidant responses, apoptosis, and PI3K/AKT/mTOR signaling in granulosa cells from women undergoing intracytoplasmic sperm injection, stratified into poor (POR), normal (NOR), and high ovarian response (HOR) groups (n = 30). Follicular fluid oxidative stress markers, antioxidant systems, and regulatory factors were analyzed, while intracellular reactive oxygen species, DNA fragmentation, and chromatin integrity were evaluated in granulosa cells. In addition, PI3K/AKT/mTOR pathway activity was assessed using immunohistochemistry and immunofluorescence. Compared with NOR, both POR and HOR groups exhibited increased oxidative stress, reflected by elevated MDA levels (p < 0.05), accompanied by divergent antioxidant responses, including reduced GSH levels in POR and a progressive increase in SOD activity from NOR to HOR (p < 0.05). Although melatonin and TGF-β levels did not differ significantly, melatonin showed a trend toward lower levels in POR and higher levels in HOR. Granulosa cell ROS production and apoptosis-related markers were significantly increased in both response extremes, particularly in POR (p < 0.05). Consistently, PI3K/AKT/mTOR signaling displayed differential regulation, with reduced activation in POR and enhanced activation in HOR compared to NOR (p < 0.05). Overall, NOR appears to reflect a balanced redox and molecular state, whereas both poor and high responses are characterized by oxidative stress, cellular damage, and dysregulated PI3K/AKT/mTOR signaling. These findings suggest that deviation from physiological equilibrium, rather than response magnitude alone, is a key determinant of granulosa cell dysfunction and impaired follicular competence.
Hydroxy-safflor yellow A (HSYA), a bioactive compound from Carthamus tinctorius, has reported anti-inflammatory and antitumor activities. However, its effects on breast cancer and the underlying immune mechanisms remain poorly defined. In this study, we investigated the immunomodulatory role of HSYA in breast cancer, with a focus on its impact on tumor-associated macrophages (TAMs) and T cell responses. Using in vitro assays, murine breast cancer models, and single-cell transcriptomic analysis, we found that HSYA significantly inhibited tumor growth and reshaped the tumor immune microenvironment. Flow cytometry and transcriptomic profiling revealed that HSYA treatment reduced tumor-associated macrophages (TAMs) infiltration and suppressed C-C motif chemokine ligand 5 (CCL5) expression, which was associated with enhanced recruitment and activation of tissue-resident memory T (TRM) cells. Co-culture and functional assays further supported a role for the TAM/CCL5 axis in mediating these immune effects. Collectively, our findings demonstrate that HSYA exerts antitumor activity, at least in part, by modulating the TAM/CCL5/TRM axis, highlighting its potential as an immunomodulatory therapeutic strategy in breast cancer.
Metabolic diseases have increased worldwide in recent decades, mainly due to a sedentary lifestyle and an unhealthy diet, with diet identified as an important regulator of gut microbiota composition. The use of natural products, such as Crocus sativus tepals extract (CTE) could be a promising approach to alleviate metabolic disorders. The aim was to investigate the potential ameliorative mechanisms of CTE in metabolic disorders induced by a high-fat diet in an animal model, focusing on the composition of the gut microbiota and its relationship with the gut-liver axis. We analyzed liver-related biochemical and morphological parameters in mice fed a 60% fat diet for 14 weeks and orally treated with CTE during the last 5 weeks of the diet. In addition, jejunal and liver histology, intestinal barrier integrity, inflammation and oxidative stress, liver inflammation and lipid metabolism were investigated. The results showed that oral administration of CTE restored the composition of the gut microbiota and specifically promoted short-chain fatty acids-producing and anti-inflammatory bacterial genera. It also improved intestinal barrier integrity and reduced inflammation in the jejunum and liver, along with a suppression of Fas and CerS6 expression in the liver and a reduction in circulating free fatty acids and β-hydroxybutyrate levels. Our results indicate a possible link between the gut microbiota and the metabolic benefits of treatment with CTE, suggesting its therapeutic potential for the prevention or treatment of metabolic disorders.
Cutaneous squamous cell carcinoma (CSCC) is a malignant tumor originating from epidermal keratinocytes. In various types of tumors, ferroptosis is a vital iron-dependent form of regulated cell death. Recent studies suggest that heat shock protein 27 (HSP27), encoded by the heat shock protein family B member 1 (HSPB1) gene, is involved in the regulation of ferroptosis, but the specific mechanism remains unclear. In this study, CSCC cell lines were transfected with lentivirus-mediated HSPB1-shRNA or lentivirus carrying overexpressed HSPB1. CSCC cell lines, xenograft mouse models, and ferroptosis inhibitors or inducers were applied to verify the mechanism and function of HSP27. Downregulation of HSP27 inhibited the proliferation, migration, and invasion of CSCC cells, whereas upregulation of HSP27 showed the opposite results. Similarly, tumor volume and weight were reduced after HSP27 was downregulated in vivo. Further studies revealed that HSP27 promoted the growth of CSCC cells and tumors by inhibiting ferroptosis, and the downregulation of HSP27 enhanced ferroptosis induced by Erastin. Ferrostatin-1 or Erastin successfully reversed the phenotype triggered by HSP27 alterations. HSP27 can induce the growth of CSCC by inhibiting ferroptosis, and is expected to become a new target for the treatment of CSCC.
Glioma, a highly aggressive brain cancer, thrives in an immunosuppressive tumor microenvironment (TME). The Unfolded Protein Response (UPR) is a key adaptive stress pathway implicated in therapy resistance. To discover clinically actionable biomarkers, we employed an integrated multi-omics strategy, combining single-cell and bulk transcriptomic data from multiple glioma cohorts. Using hdWGCNA network analysis, we identified a UPR-associated gene module that defined two molecular subtypes with stark survival differences. Through a rigorous machine learning pipeline (Random Survival Forest, LASSO, and CoxBoost), ANXA5 emerged as the central prognostic gene. High ANXA5 expression consistently predicted poorer overall survival across eight independent datasets, validating its robust prognostic value. Functionally, ANXA5 was linked to extracellular matrix remodeling and immune modulation. Multi-omics profiling revealed that ANXA5-high gliomas exhibit a T-cell-inflamed yet immunosuppressive TME, characterized by elevated immune checkpoint expression. Crucially, ANXA5 demonstrated strong predictive power for response to immune checkpoint blockade (ICB), showing significant correlation with nine established immunotherapy response signatures and accurately discriminating responders from non-responders in six independent ICB-treated clinical cohorts (AUC: 0.65-0.78). Genomic analysis associated ANXA5 expression with distinct mutation patterns (EGFR/PTEN vs. IDH1/TP53). In vitro knockdown of ANXA5 confirmed its oncogenic role, as it suppressed glioma cell proliferation and invasion. Our study establishes ANXA5 as a prime example of a translatable biomarker discovered through multi-omics integration. It functions dually as a prognostic indicator and a predictive biomarker for immunotherapy, offering a tangible framework for patient stratification and personalized therapeutic strategies in glioma, thereby bridging a critical gap toward clinical translation.
Senescent cells are characterized by the up-regulation of senescence markers and exhibit key features, such as irreversible growth arrest and the senescence-associated secretory phenotype (SASP), which is mainly regulated by transcription factors of nuclear factor κB (NF-κB). Lipocalin-2 (LCN2), a glycoprotein secreted by immune cells, astrocytes, and epithelial cells, is present in saliva and gingival crevicular fluid and possesses antimicrobial and immunomodulatory properties. Although LCN2 expression is mainly regulated by NF-κB, the effects of aging and cellular senescence on salivary LCN2 protein concentrations remain unknown. We herein demonstrated that LCN2 protein levels in the serous acinar cells of salivary glands, oral epithelial cells, and saliva were higher in aged mice than in young mice. However, in primary oral keratinocytes and salivary gland epithelial cells, replicative senescence and DNA damage-induced senescence did not increase LCN2 expression, with similar results being obtained for the SASP factors tumor necrosis factor-alpha and interleukin-1β (IL-1β). Although the cyclic GMP-AMP synthase-mediated induction of LCN2 expression has been reported in astrocytes, its expression decreased with cellular senescence, and its ligand did not induce LCN2 expression in these oral-related epithelial cells. Conversely, an IL-1β treatment significantly induced LCN2 expression and secretion, even in senescent epithelial cells. The source of IL-1β was not senescent fibroblasts, but M1 macrophages that accumulate with inflammaging. Collectively, these results suggest that aging up-regulates LCN2 expression in oral-related epithelial cells mainly via IL-1β secreted from M1 macrophages, rather than through the induction of their senescence.
Electrical stimulation (ES) has emerged as a promising technique in the field of bioengineering and biomedicine, particularly in bone regeneration and cell differentiation. ES using alternating current (AC) is based on the periodic reversal of current direction, which generates oscillating electric fields. The application of an electric field has effects on cell growth and differentiation, as well as on morphology and migration. This study aimed to explore the effect of applying AC electrostimulation within the proliferation, differentiation, and morphology process of osteoblastic cells. The electrical stimulation signals were daily applied for 3 h during 14 days. Different frequencies were tested (1 Hz, 10 Hz, 100 Hz, and 1 kHz), with amplitudes of 125, 250, 500, 750, 1000, and 1500 mV/mm. Cell viability was estimated using the AlamarBlue, and MC3T3-E1 differentiation levels were evaluated through alkaline phosphatase (ALP) activity. RUNX2, OSX, ALP, OPG, and RANKL gene expression was assessed by RT-PCR. Morphological analysis was performed through cell transfection followed by immunofluorescence. Statistical analysis was conducted by SPSS.23 and graphs generated through Graph-pad. Viability and ALP activity were optimal at 10 Hz. Once the frequency was defined, RUNX2, OSX, ALP, OPG, and RANKL gene expression revealed an increase in the differentiation and osteogenic activity levels at 10 Hz and 500-750 mV/mm. As well as, morphological studies showed an increase in the area, pseudopodia length, and numbers at 500 mV 10 Hz conditions. The optimal ES condition to differentiate MC3T3-E1 cells is 10 Hz 500-750 mV/mm. Electrostimulation has emerged as a promising technique in the field of bioengineering and biomedicine, particularly in bone regeneration and cell early maturation.
Researchers have developed hybrid bionic platforms for odor detection, inspired by natural chemoreceptive systems, advancing artificial olfactory systems that recognize specific volatile compounds. Odorant binding proteins (OBPs) are small carrier proteins found in the olfactory organs of mammals and insects. When coupled with electrical transducers, OBPs act as recognition elements, converting chemical signals into electrical outputs. This enables the development of biological electronic noses that are based on biomimetics and aim for sustainability. The objective of this review is to provide a comprehensive and updated overview of OBP-based biosensors, with a particular focus on insect OBPs as biorecognition elements, and to critically examine their applications, advantages, and technological potential across different fields. OBP-based biosensors show strong promise in medical diagnostics, environmental monitoring, food quality, insect pest control, and security. Insects demonstrate remarkable sensitivity to specific odors which makes them excellent models for designing bioinspired biosensors. Compared to conventional methods, OBP-based biosensors offer significant advantages in terms of portability, rapid response, and cost-effectiveness. OBPs are remarkably stable under different environmental conditions and can bind both volatile and aqueous-phase molecules, enhancing their functional versatility. Moreover, they can be produced through biotechnological processes using renewable resources, supporting eco-friendly innovation. These advantages make OBPs ideal candidates for next-generation biosensors in fields requiring real-time and on-site chemical detection.
Myocardial infarction (MI) is a major cause of mortality. This study explores the cardioprotective effects of Coenzyme Q10 (CoQ10) and its molecular mechanisms in MI. A rat myocardial ischemia-reperfusion model was established, and infarct size was assessed via triphenyl tetrazolium chloride (TTC) staining. Hematoxylin and eosin (H&E) and Masson's staining analyzed histological changes, while terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) and flow cytometry assessed apoptosis. Western blot and immunofluorescence evaluated collagen I, collagen III, α-SMA, histone deacetylases (HDACs) (4,5,7,9,11), O-GlcNAc transferase (OGT), O-GlcNAcase (OGA), and O-GlcNAcylation. Enzyme-linked immunosorbent assay (ELISA) measured tumor necrosis factor (TNF)-α, interleukin (IL)-6, transforming growth factor (TGF)-β, suppression of tumorigenicity (ST)-2, galectin (Gal)-3, and growth differentiation factor (GDF)-15, while cell counting kit-8 (CCK-8), EdU, and Transwell migration assays assessed fibroblast proliferation and migration. O-GlcNAcylation of HDAC4 was analyzed using Click-iT labeling and immunoprecipitation. Cycloheximide chase assay and ubiquitination assay detected the impact of O-GlcNAcylation at Ser455 on HDAC4 protein stability. CoQ10 significantly improved cardiac function in rat models, reducing infarct size, apoptosis, fibrosis, and inflammation. CoQ10 treatment in MIRI rats increased ejection fraction (EF) and fractional shortening (FS), reduced infarct volume (TTC staining), and decreased collagen deposition (Masson's, Sirius Red staining). TUNEL and flow cytometry confirmed reduced cardiomyocyte apoptosis, while ELISA showed lower TNF-α, IL-6, and TGF-β, indicating suppressed inflammation. At the molecular level, CoQ10 reduced global O-GlcNAcylation together with decreased OGT and increased OGA expression. Western blot and immunofluorescence confirmed reduced HDAC4 levels, linked to O-GlcNAcylation suppression particularly at Ser455. CoQ10 promoted the degradation of HDAC4 through the precise inhibition of its stabilizing O-GlcNAc modification at Ser455. In vitro, CoQ10 enhanced oxygen-glucose deprivation and reperfusion (OGD/R) cardiomyocyte survival, reduced apoptosis (TUNEL, flow cytometry), and suppressed HDAC4 expression. In fibroblasts, CoQ10 inhibited TGF-β1-induced proliferation/migration (CCK-8, EdU, Transwell assays) and reduced collagen I, collagen III, and α-SMA expression, indicating anti-fibrotic effects. Furthermore, HDAC4 knockdown in vivo improved cardiac function, and in vitro reduced cardiomyocyte apoptosis and fibroblast proliferation/migration, reinforcing HDAC4's role in MI pathology. CoQ10 exerts cardioprotective effects by modulating O-GlcNAc modification and HDAC4 expression.
Trifuhalol A (TFA), a phlorotannin derived from the edible brown seaweed Agarum cribrosum, has been reported to exert diverse physiological activities, yet its anti-diabetic mechanism remains unclear. This study systematically investigates the multi-targeted anti-diabetic effects of TFA, with a particular focus on enhancing glucose uptake and protecting pancreatic islets. In vitro enzyme inhibition assays demonstrated that TFA significantly inhibited the activities of α-glucosidase and α-amylase, indicating its potential to attenuate postprandial glycemic excursions by modulating carbohydrate hydrolysis. Additionally, TFA effectively suppressed the formation of advanced glycation end-products (AGEs), potentially reducing the risk of diabetes-associated complications. Mechanistically, TFA enhanced glucose uptake in C2C12 myotubes by activating the PI3K/Akt and AMPK signaling pathways, which in turn promoted the translocation of glucose transporter type 4 (GLUT4) to the plasma membrane, thereby facilitating cellular glucose utilization and insulin sensitivity. Furthermore, in vivo investigations using an alloxan-induced type 1 diabetic zebrafish further confirmed the bioefficacy of TFA, as evidenced by its capacity to reduce hyperglycemia, alleviate oxidative stress, and protect pancreatic islets, without eliciting observable systemic toxicity. Taken together, these findings provide both mechanistic and functional evidence supporting TFA as a safe and potent multi-target bioactive compound with promising applications in the development of functional foods and therapeutic strategies for diabetes management.
Bile acids (BAs), long recognized for their role in lipid digestion, have recently emerged as key signaling molecules at the interface of host metabolism, immunity, and gut microbiota (GM). BAs are synthesized in hepatocytes and subsequently extensively modified by microbial enzymes in the gut, producing a diverse and dynamic pool that strongly shapes the GM-immune axis. Through activation of receptors such as the Farnesoid X receptor and the G protein-coupled receptor TGR5, BAs regulate inflammation, metabolic pathways, and intestinal immune homeostasis, particularly influencing the balance between regulatory T cells and pro-inflammatory Th17 cells. Microbial transformations, primarily deconjugation and 7α-dehydroxylation, further diversify BA species, modulating receptor affinities and immunoregulatory functions. Dysbiosis-associated alterations in these processes contribute to the pathogenesis of inflammatory disorders, including inflammatory bowel disease (IBD). Consequently, BAs are increasingly recognized as promising biomarkers for monitoring disease activity and predicting therapeutic response, although validation in standardized, prospective cohorts remains necessary. Recent advances in high-resolution analytical techniques, notably high- and ultra-performance liquid chromatography coupled with tandem mass spectrometry (HPLC- and UPLC-MS/MS), have enabled precise, high-throughput quantification of BA species in serum and fecal samples. These methods both deepen mechanistic understanding of BA-mediated immunomodulation and support the development of GM- and BA-targeted therapies. This review emphasizes the central role of BAs in GM-immune axis regulation, delineates their complex interplay with host and microbial factors, and surveys evolving analytical strategies that facilitate their study in health and disease.
Plant-based food products have been developed from diverse plant sources as new food choices. Fermentation of plant matrices with lactic acid bacteria (LAB) has been shown to improve quality and bioactivity of the resulting product while proteolysis by the LAB in the plant-based matrix remains to be elucidated. In this study, a hazelnut-based matrix prepared for a plant-based product was fermented with four different starter cultures of LAB, and their effects on proteolysis, bioactivity and allergenicity were investigated. Sucrose supplementation of the hazelnut matrix stimulated fermentation and time to reach to the target pH of 4.5 was shortened. CH-1 was the fastest acidifying culture reducing pH to the target value after 5 h. While the cultures RSF-736 and CHN-11 required 18 h for fermentation, R-707 was co-cultured with CH-1 to reach the target pH within the same time. Bacterial counts were in the range of 5-8 log cfu/g without a significant change after 15 days of storage in the hazelnut-based products. Level of proteolysis as measured by changes in soluble protein and total free amino acid contents differed among the cultures. Reductions in the amounts of hazelnut proteins were also confirmed by SDS-PAGE analysis, especially in the products prepared with cultures R-707+CH-1 and RSF-736. Allergenicity of the hazelnut matrix, determined by a hazelnut-specific ELISA test, significantly decreased after fermentation with all the cultures. Fermentation also enhanced total phenolic content and antioxidant activity of the hazelnut matrix with CHN-11 demonstrating the highest values after storage. On the other hand, fermentation did not significantly alter α-amylase inhibitory activity compared to the activity of 10.2% in the unfermented control. In addition, fermentation resulted in no change or a slight reduction in ACE inhibitory activity compared to the activity of 46.9% in the unfermented control depending on the culture. These findings demonstrate that LAB species can degrade hazelnut matrix leading to a reduction in allergenicity and enhancement of antioxidant activity.
Accumulating evidence suggests that SUMOylation plays a crucial role in the progression and resistance of secondary hyperparathyroidism (SHPT). However, the precise mechanism of SUMOylation in SHPT remains unclear. We identified the potential role of SUMOylation in SHPT based on RNA sequencing data obtained from Gene Expression Omnibus (GEO) datasets. Clinical samples were used to verify the expression of SENP1, SUMO1, SUMO2, GRHL2, and vitamin D receptor (VDR) in SHPT cells and tissues. Primary cells were extracted for subsequent experiments. Plasmid transfection and small interfering RNA (siRNA) were used to modulate SENP1 expression in SHPT primary cells. VDR relative expression was detected by Western blot (WB) and immunofluorescence. The effects of SENP1 on SHPT cell apoptosis and anti-proliferation were analyzed by flow cytometry and WB. Co-immunoprecipitation (Co-IP), chromatin immunoprecipitation (ChIP), and ChIP-quantitative polymerase chain reaction (ChIP-qPCR) were employed to explore the regulatory mechanisms of SENP1 in SHPT. We found that SUMOylation was significantly upregulated in SHPT and was closely related to calcitriol resistance. SENP1 and GRHL2 were downregulated. SENP1 was found to upregulate VDR and downregulate the SUMOylation of VDR, which mediates SENP1's regulation of SHPT cell apoptosis and anti-proliferation. Mechanistically Co-IP assays revealed binding between VDR, SENP1, SUMO1, and SUMO2. ChIP assays indicated that transcription of SENP1 was regulated by GRHL2, with binding sites identified by ChIP-qPCR. Additionally, we identified the potential binding pocket of SENP1 and screened 10 candidate small-molecule drugs approved by the US Food and Drug Administration (FDA). Our findings indicate a distinct mechanism of SENP1-mediated VDR SUMOylation and establish the critical role of the GRHL2/SENP1/VDR signaling axis in SHPT development.
Vestigial-like family member 3 (VGLL3), a transcriptional cofactor of the TEA domain family, has been previously identified as a regulator of osteoblast differentiation. Building upon our previous findings, we investigated VGLL3 function in MC3T3-E1 osteoblasts using an integrated approach combining transcriptomic analysis and functional assays to identify its downstream effectors and explore associated autophagy mechanisms. RNA-seq analysis of Vgll3-knockdown (shVgll3) cells identified death-associated protein kinase 2 (DAPK2), a regulator of autophagy, as a downstream effector. Autophagic activity was examined using transmission electron microscopy and western blot analysis of LC3-II and p62 proteins. The effects of Dapk2 knockdown (shDapk2) on osteoblast differentiation were evaluated using qPCR, western blotting, alkaline phosphatase staining, and Alizarin Red staining. Rapamycin treatment was used to determine whether pharmacologic activation of autophagy could restore osteoblast function. Vgll3 knockdown significantly suppressed autophagic flux, as evidenced by fewer autophagic vacuoles, decreased LC3-II accumulation, and increased p62 expression. A comparable reduction in autophagic activity was observed in shDapk2 cells and was accompanied by impaired osteoblast differentiation. Rapamycin treatment partially restored autophagy and osteogenic differentiation in Vgll3-deficient cells. Finally, overexpression of DAPK2 partially rescued autophagic activity and osteogenic differentiation in shVgll3 cells, supporting its role as a key downstream functional effector. FOXM1 was further implicated as a potential transcriptional regulator contributing to DAPK2 expression. Collectively, our findings suggest that VGLL3 may influence osteogenic differentiation in osteoblasts, potentially involving DAPK2-associated autophagy.