共找到 20 条结果
Autophagy is increasingly understood as a lipid-governed membrane program rather than a solely protein-driven degradative pathway. This review combines molecular and mechanistic evidence showing how lipid molecules, metabolic enzymes, and membrane physical properties coordinate autophagy from induction to lysosomal degradation. We highlight phosphoinositide microdomains and their cognate kinases and phosphatases as spatial cues that nucleate phagophores, control maturation, and regulate lysosome reformation. We also discuss alternative phosphoinositide sources, sphingolipid and ceramide signaling, phosphatidic acid and diacylglycerol metabolism, fatty-acyl composition, and acyl-CoA signaling as determinants of membrane curvature, tension, leaflet asymmetry, and phase behavior in autophagy. Key protein effectors and their lipid binding motifs and domains are integrated into a model in which lipid chemistry and mechanics gate enzymatic activities. We present integrated lipid-protein-biophysics approaches to highlight outstanding questions and uncover predictive principles.
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.
Despite its aggressiveness and unfavorable prognosis, the key molecular drivers of Hepatocellular carcinoma (HCC) are still not fully defined. The role of ribosomal protein S8 (RPS8) in HCC development and its regulatory mechanisms received little attention. We investigated the role of RPS8 in HCC progression by delineating its biological functions, clinical relevance, and mechanistic underpinnings. We conducted comprehensive multi-omics analysis using public datasets (TCGA, GEO) to evaluate RPS8 expression and its prognostic value in HCC. Single-cell RNA-seq was utilized to map the distribution of RPS8 expression within the tumor microenvironment. The biological functions of RPS8 were validated through a series of in vitro assays (e.g., colony formation, Transwell, wound healing) and in vivo subcutaneous xenograft models using RPS8 knockdown and overexpression systems. Whole-transcriptome sequencing was performed to identify the downstream pathways regulated by RPS8. RPS8 was elevated in HCC at both the transcriptional and protein levels, and its overexpression independently signaled poor prognosis. In functional assays, RPS8 increased HCC cell proliferation, migration, and invasion in vitro and stimulated tumor growth in vivo. This pro-tumorigenic activity was associated with the induction of epithelial-mesenchymal transition (EMT) and angiogenesis. Mechanistically, we revealed that RPS8 exerts these effects by activating the endoplasmic reticulum (ER) stress pathway. Furthermore, high RPS8 expression was associated with a remodeled tumor immune microenvironment, characterized by increased CD8+ T cell infiltration. Findings identify RPS8 as a critical oncogene that promotes HCC progression via activation of ER stress. RPS8 can be leveraged as a prognostic biomarker and as a potential focus of therapy in HCC.
Ferroptosis is a highly synchronized form of non-apoptotic intracellular iron-dependent cell death which is regulated by multiple cellular metabolic pathways. Extensive studies suggest that ferroptosis could efficiently set the therapy resistant cancer cells on the road to ruin, thus providing new opportunities for cancer therapy. And-1 is an acidic nucleoplasmic DNA binding protein which plays a vital role in DNA replication and repair. Here in this study, we report a novel function of And-1 in regulating erastin-induced ferroptosis in ovarian cancer cells. And-1 overexpression (OE) enhanced erastin-induced ferroptosis by modulating the level of ferroptosis biomarkers such as MDA, GSH, Fe2+ and lipid peroxidation; while knockdown (KD) of And-1 produces opposite results. Moreover And-1(OE) suppressed the expression of NRF2, SLC7A11, and FTH1 and promoted the expression of TFR1 and ATF3 while And-1 (KD) produces opposite results. Further mechanistic study revealed that And-1 inhibits the expression of SLC7A11 by increasing ATF3 expression. Moreover, using pharmacological inhibitors, we have shown that erastin induces ferroptosis through multiple mechanisms. Taken together, our data suggest that And-1 enhanced erastin-induced ferroptosis in OC cells by inhibiting system xc - expression through ATF3 and suppressing NRF2 expression.
Paclitaxel (PTX) resistance limits cervical cancer therapy. Ferroptosis suppression via GPX4 and redox remodeling has emerged as a resistance mechanism, but upstream regulators remain unclear. To determine whether ANO6 drives PTX resistance by inhibiting ferroptosis and to define the ANO6-GPX4 axis mechanistically and therapeutically. Transcriptomic analyses, immunohistochemistry on clinical specimens, and cervical cancer cell models with gain/loss of ANO6 were combined with ferroptosis assays, mitochondrial imaging, apoptosis/viability assays, and Co-IP/CHX chase to assess ANO6-GPX4 interaction and stability. GPX4 transcriptional control was probed by ChIP-qPCR and dual-luciferase. PTX sensitivity was tested in vitro and in xenografts, with or without the ferroptosis inducer RSL3. ANO6 was overexpressed in cervical cancer and associated with worse prognosis. ANO6 knockdown reduced GPX4, SLC7A11, and NRF2, increased ACSL4, elevated lipid peroxidation and iron load, disrupted mitochondrial integrity, and heightened PTX cytotoxicity; ANO6 overexpression had opposite effects. ANO6 physically associated with GPX4 and preserved its protein stability; NRF2 enhanced GPX4 promoter activity, supporting a dual (post-translational/transcriptional) maintenance of GPX4 under ANO6 control. In PTX-resistant cells, ANO6 was upregulated; its depletion restored ferroptosis and PTX sensitivity, whereas GPX4 overexpression rescued resistance. In vivo, ANO6 overexpression promoted tumor growth and PTX resistance, while PTX + RSL3 synergistically suppressed tumors and reversed GPX4-axis signaling. ANO6 confers PTX resistance by sustaining GPX4-dependent ferroptosis evasion and mitochondrial homeostasis. Targeting the ANO-GPX4 axis, alone or combined with ferroptosis induction, may improve chemotherapy sensitivity in cervical cancer.
In search for novel antibiotic candidates, polyphenols represent an unexploited source with a nearly inexhaustible number of promising compounds. In addition to the antimicrobial activity, some polyphenols modulate the immune response in a way that could enhance the clearance of invading pathogens. This study investigated four resveratrol-derived dimeric compounds, dehydro-δ-viniferin, (±)-trans-δ-viniferin, viniferifuran, and (±)-ε-viniferin, for their antimicrobial activity against methicillin-resistant S. aureus. In addition, their immunomodulating properties in bacterially challenged dendritic cells were assessed. We identified dehydro-δ-viniferin as exhibiting the most potent antimicrobial activity against methicillin-resistant Staphylococcus aureus with a minimal inhibitory concentration of 2 μg/mL and a minimal bactericidal concentration of 8 μg/mL, respectively. Furthermore, dehydro-δ-viniferin showed significant enhancement of the S. aureus-induced IL-12 production from bone marrow-derived dendritic cells, which could be ascribed to a decrease in the production of IL-10. Only if dehydro-δ-viniferin was administered to dendritic cells prior to bacterial stimulation, IL-12 production was increased; if added later, the increase in IL-12 was lost although the IL-10 production was reduced. This suggests that dehydro-δ-viniferin interferes with the signaling pathway leading to IL-10, which in turn influences the IL-12 production in a time dependent manner. Collectively, dehydro-δ-viniferin is a potent antimicrobial against methicillin-resistant S. aureus with enhancing effects on the bacterially induced IL-12 response in antigen-presenting cells. We therefore suggest dehydro-δ-viniferin as a promising antibiotic candidate against multidrug-resistant Gram-positive bacteria.
Glioblastoma is lethal brain tumor with dismal prognosis. ETV1, an ETS family transcription factor, has been implicated in multiple malignancies, yet its precise role and regulatory mechanisms in glioma progression remain incompletely defined. This study investigated the role of ETV1 in maintaining the malignant phenotype of high-grade gliomas through a novel regulatory axis involving microRNA-3175 (miR-3175) and STEAP2. ETV1 expression was analyzed in TCGA glioma datasets (n = 692) and 85 patient tissue specimens using bioinformatics, quantitative RT-PCR, Western blotting, and immunohistochemistry. ETV1 function was assessed through gain-of-function and loss-of-function studies in glioma cell lines (U251, A172) and validated in nude mouse xenografts with bioluminescence imaging. The ETV1-miR-3175-STEAP2 regulatory axis was characterized using chromatin immunoprecipitation, luciferase reporter assays, biotin-streptavidin pulldown, RNA immunoprecipitation, and rescue experiments. ETV1 was significantly upregulated in gliomas, with expression correlating to tumor grade and reduced overall patient survival. ETV1 overexpression promoted glioma cell proliferation, migration, and invasion in vitro and enhanced tumor growth while reducing survival in vivo, with elevated bioluminescence radiance indicating enhanced tumorigenicity, whereas ETV1 knockdown produced opposite effects and diminished bioluminescent signals. ETV1 transcriptionally activated miR-3175 through direct binding to its promoter. miR-3175 directly targeted and suppressed STEAP2 expression via conserved 3'UTR binding sites. STEAP2 functioned as a tumor suppressor; overexpression inhibited malignant phenotypes. Rescue experiments confirmed miR-3175 and STEAP2 mediate ETV1-driven glioma progression through a functional ETV1/miR-3175/STEAP2 regulatory axis. This study revealed a novel ETV1/miR-3175/STEAP2 regulatory axis driving glioma tumorigenesis and identified potential therapeutic targets for intervention.
Dextromethorphan (DXM), a widely used antitussive agent, was investigated for its effects on mitochondrial F1FO-ATPase activity and oxidative phosphorylation. Our results demonstrate that DXM inhibited F1FO-ATPase independently of the thiol redox state. Mutual exclusion analysis highlighted an overlapping binding site between DXM and dicyclohexylcarbodiimide (DCCD), indicating a shared or adjacent binding site in the membrane-embedded FO domain of the enzyme. These findings suggested that DXM selectively targeted the proton translocation mechanism of F1FO-ATPase during the ATP hydrolysis and synthesis of ATP. Moreover, kinetic analysis confirmed a high affinity of DXM for the enzyme, with an inhibitory efficiency of 2.37 mM-1⸱s-1. Importantly, DXM did not affect electron transport chain activity but impaired ATP synthesis, as evidenced by altered respiratory control ratios of oxidative phosphorylation. The data obtained offer new insights into its off-target mitochondrial effects and potential implications for bioenergetic regulation.
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.
This review systematically explores the dynamic regulatory roles of lysine lactylation (Kla) in the tumor microenvironment (TME) and its clinical translational potential. As an emerging post-translational modification, Kla modifies histones and non-histone proteins via lactate generated by the Warburg effect, thereby reshaping tumor metabolism and immune landscapes. Mechanistically, Kla orchestrates metabolic reprogramming and immunosuppression through key signaling pathways such as HIF-1α, mTOR, and NF-κB. Specifically, it promotes the activation of immunosuppressive cells while inhibiting cytotoxic CD8+ T cells and NK cells, fostering tumor immune escape. Preclinical studies demonstrate that targeting lactate metabolism or lactylation enzymes restores immune effector functions and enhances immune checkpoint therapy efficacy. However, challenges such as tumor heterogeneity, metabolic plasticity, and systemic toxicity remain. Future research should focus on Kla's crosstalk with other epigenetic modifications, spatiotemporal dynamics in TIME, and clinical translation to unlock its potential as a biomarker and precision oncology target.
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.
Major depressive disorder (MDD) is a highly disabling psychiatric illness characterized by persistent low mood and psychomotor retardation, often leading to cognitive impairment or even suicidality. As the pathogenesis remains poorly understood, the currently available treatment regimens are mostly symptomatic therapies with little satisfactory curative effect. The emerging paradigm of gut-brain axis has highlighted gut dysbiosis as a key etiological factor to elicit neuro-inflammation and jeopardize the central nervous system homeostasis. In this study, we evaluated the ameliorating effects of flaxseed lignans on the gut microbiome to regain the gut micro-environmental functionality and alleviate MDD. The lignans significantly mitigated the severity of disease and markedly altered the gut microbiota structure in the participants. In the MDD mouse model, the lignans reversed the experimental depression-like behaviors, repaired neural and gut damage and restored barrier integrity. Of great significance, the lignans elevated the levels of 5-hydroxytryptamine (5-HT), brain-derived neurotrophic factor (BDNF) and γ-aminobutyric acid (GABA), and markedly attenuated microglial and systemic inflammation. Mechanistically, the lignans inhibited the IL-17/AP-1/NF-κB axis through direct interaction with the Fos protein. In the BV-2 cells, the lignan enterolactone reduced levels of nitric oxide and pro-inflammatory cytokines, further validating the anti-inflammatory mechanism of the lignans. Together, these findings demonstrate that lignans exert potent antidepressant effects by modulating the gut-brain axis and resolving neuro-inflammation, providing useful information for the development of novel therapeutic strategies for MDD prevention and treatment.
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.
Naringenin (NAR) and hesperetin (HES) are citrus flavanones that have been shown to exert various geroprotective effects. However, key regulators of these processes remain unexplored. To address this gap, we investigated the mechanisms through which NAR and HES modulate hepatic redox control, persulfidation, and senescence-associated pathways in 24-month-old male Wistar rats. Animals were treated orally with NAR or HES (15 mg/kg b.w.) for 4 weeks, while control groups remained physiologically intact or received vehicle. Liver tissue was analyzed using a dimedone-based switch method to evaluate overall protein persulfidation (P-SSH), Western blot, qPCR, and histological analysis, including quantification of IHC staining and IF labeling. Our results showed that both citrus flavanones increased total P-SSH levels while exerting distinct regulatory effects on the H2S metabolic network. Only HES upregulated gene expression of H2S-producing enzymes (cystathionine beta-synthase CBS, cystathionine gamma-lyase CSE, and mercaptopyruvate sulfurtransferase MST) and catabolic enzymes (sulfide:quinone oxidoreductase SQR and thiosulfate sulfurtransferase TST), together with CBS protein, whereas NAR decreased CSE protein expression. Both NAR and HES treatment activated the Nrf2 signaling pathway, as evidenced by increased nuclear translocation of Nrf2 in the treated groups. Concurrently, NAR and HES increased Sirt1 expression, which was accompanied by decreases in p16 and β-galactosidase expression, indicating attenuation of hepatic senescence. Together, these findings highlight a synergistic interplay among persulfidation, H2S metabolism, and Nrf2/Sirt1 signaling as a key mechanism by which citrus flavanones support hepatic resilience during aging.
Pancreatic cancer (PC) remains a highly lethal malignancy with limited treatment options, largely due to its heterogeneity and therapy resistance. While ferroptosis-a form of iron-dependent cell death driven by lipid peroxidation-has emerged as a relevant pathway, its role in PC is incompletely understood, as is the oncogenic function of Centrosomal Protein 55 (CEP55). Here, we integrated TCGA data with immunohistochemical validation and demonstrated that CEP55 is significantly overexpressed in PC and correlates with advanced disease and poor prognosis. Functionally, CEP55 knockdown suppressed proliferation, migration, and clonogenicity, while inducing ferroptosis, as evidenced by elevated lipid peroxidation, iron accumulation, and glutathione depletion. Mechanistically, CEP55 silencing downregulated key ferroptosis suppressors, including GPX4, SLC7A11, and NQO1, increasing cellular sensitivity to ferroptotic stress. Erastin, a ferroptosis inducer, enhanced ferroptosis in CEP55-deficient cells and counteracted the tumor-promoting effects of CEP55 overexpression. In vivo, CEP55 silencing reduced tumor growth and altered ferroptosis markers. Our findings establish CEP55 as a novel driver of PC progression via ferroptosis suppression, supporting its potential as both a prognostic biomarker and a therapeutic target for combination strategies aimed at overcoming PC resistance.
Circular RNAs (circRNAs), a class of non-coding RNAs with a closed-loop structure, have emerged as a focal point in cancer research due to their stability, tissue specificity, and high abundance in body fluids. This paper systematically elaborates on the dual roles of circRNAs in cancer. Specifically, they can act as tumor suppressors to inhibit tumor progression or as oncogenes to promote tumorigenesis. Their core mechanisms involve epigenetic regulation, including modulation of DNA methylation, histone post-translational modifications, and functioning as miRNA sponges to affect target gene expression. CircRNAs regulate cancer hallmarks through these mechanisms, such as cell proliferation, apoptosis resistance, angiogenesis, and invasion-metastasis. Additionally, due to their unique biological properties, circRNAs show great potential in cancer diagnosis and prognostic evaluation. Intervention strategies targeting the circRNA-epigenetic axis, such as antisense oligonucleotides and CRISPR technology, provide new directions for cancer therapy. This study aims to deeply analyze the complex regulatory networks of circRNAs in tumorigenesis and progression, explore key challenges in their translation from basic research to clinical practice, and offer theoretical basis and practical references for understanding epigenetic dysregulation in cancer and developing novel diagnostic and therapeutic approaches.
Resveratrol (RSV; 3,5,4'-trihydroxy-trans-stilbene) is a natural polyphenolic compound with notable antioxidant, anti-inflammatory, and immunomodulatory properties. It has been investigated for therapeutic applications in cardiovascular disease, cancer, and neurodegenerative disorders. This review emphasizes the potential of RSV in oncology and neuroprotection, synthesizing evidence from systematic database searches and experimental studies. Despite promising biological activities, RSV is limited by poor stability, low aqueous solubility, rapid metabolism, and restricted bioavailability, necessitating improved delivery strategies such as nanoencapsulation, nanocrystals, prodrugs, and structural analogues. Mechanistically, RSV exerts anticancer and neuroprotective effects through modulation of p53, STAT3, NF-κB, and mitochondrial-mediated apoptosis. Its antioxidant actions involve regulation of reactive oxygen species (ROS), activation of NRF2, AMPK signaling, and SIRT1. RSV and related antioxidants act on multiple molecular pathways, including TP53, β-catenin, STAT3, NF-κB, NRF2-AMPK, PI3K/AKT, and SIRT1, to regulate inflammation and cell death. The balance between oxidative and antioxidative processes is critical for therapeutic efficacy. Notably, RSV-induced ROS-mediated cell death, particularly in the context of TP53 mutations, represents a promising target for future interventions. Overall, RSV demonstrates multi-target potential for cancer and neurodegenerative disease therapy, though optimization of its pharmacological profile remains essential.
Anxiety-related disorders are among the most prevalent mental health conditions worldwide, driving interest in novel, plant-based interventions. This study aimed to evaluate the antioxidant activity and capacity to alleviate anxiety-like behavior of a commercially standardized green rooibos (Aspalathus linearis) powdered extract (GRE) in vivo. The GRE was chemically characterized using liquid and gas chromatography. Antioxidant capacity (survival and ROS levels upon oxidative stress) and anxiety-like behavior reduction (octanol avoidance test) were evaluated in the Caenorhabditis elegans model, and additionally in adult zebrafish (Danio rerio) by a Novel Tank Test after 4 months of GRE ingestion. Analytical results of GRE confirmed the presence of dihydrochalcones (aspalathin, nothofagin) and flavones (orientin, isoorientin, isovitexin) as the major phenolic constituents. In C. elegans, GRE supplementation improved survival under oxidative stress conditions and significantly reduced intracellular ROS levels in a dose-dependent manner. Additionally, GRE exposure mitigated anxiety-like behavior in nematodes subjected to serotonin-deficiency conditions induced by starvation. GRE-fed fish exhibited a significant increase in time spent in the upper zone of the tank, reduced latency to reach this zone, and fewer individuals displaying bottom-dwelling behavior, while locomotor parameters (speed and distance traveled) remained unaffected, indicating reduced anxiety-like behavior levels. Data support the potential of GRE to reduce anxiety-related behavior in both invertebrate and vertebrate models, possibly mediated by its antioxidant properties and polyphenolic profile. These findings underscore the potential of A. linearis as a functional botanical candidate for future dietary interventions for anxiety support.
L-dopa decarboxylase (DDC) is the biosynthetic enzyme of dopamine and serotonin. Although DDC has been originally studied for its role in neurotransmission, it has also been detected in peripheral organs, where it is implicated in cellular homeostasis. DDC has been identified by our research team as a negative regulator of dengue virus (DENV) replication in liver cells. The latter has been attributed, at least in part, to the physical interaction of DDC protein with phosphatidylinositol 3-kinase (PI3K), and its biosynthetic function. PI3K/AKT signaling and cell survival are manipulated by DENV to favor its replication. Based on the above, we investigated whether DDC exerts its antiviral activity against DENV propagation through modulation of DENV-induced cell death and especially apoptosis. Specifically, DDC silencing in Huh7.5 cells (shDDC) significantly reduced virus-induced cytopathic effect compared to the control cells (shControl). This finding was accompanied by suppression of both early and later stages of apoptosis in the silenced cells, as shown by Annexin V/PI staining and TUNEL assay, respectively. Accordingly, upon infection, shDDC cells showed suppressed activation of key caspases, BCL-2 family members and TRAIL-receptor genes that modulate the apoptotic cascade, compared to the control cells. Moreover, mitochondrial analysis in DENV-infected cells revealed that, upon DDC silencing, a less pronounced disruption of mitochondrial membrane potential and network integrity, higher respiratory capacity, lower ROS production, and reduced cytochrome c release were observed. As the PI3K/AKT pathway is known to be affected by both DENV and DDC, next we assessed whether DDC is involved in the virus-induced apoptosis through this axis. For this, we quantified the reduction of p-AKT and p-mTOR levels caused by DENV infection in the two cell lines, which was found greater in the shControl cells. Finally, chemical inhibition of AKT phosphorylation abolished the differences in cell viability and apoptosis between the two cell lines. In total, our findings highlight the suppressive role of DDC against DENV replication by modulating the PI3K/AKT-dependent apoptotic signaling.
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.