Cyclin-dependent kinase 1 (CDK1) is a key regulator of cell cycle progression and a potential therapeutic target for invasive malignancies. However, developing selective CDK1 inhibitors with manageable toxicity remains a significant challenge. In this study, a novel series of 1,2,4-triazolobenzene sulfonamide derivatives were designed and synthesized based on the structure of JNJ7706621 and its derivative 3n, and subjected to comprehensive bioactivity evaluation and structure-activity relationship discussion. Among them, 11l emerged as a highly promising lead compound, exhibiting nanomolar inhibitory activity against CDK1 (IC50 = 5.5 nM) with high selectivity over CDK2, Aurora A, and CDK4, showing selectivity indices of 4.7-, 14.1-, and 73.2-fold, respectively. In vitro, 11l exhibited broad antiproliferative activity, particularly against HCT116 colon cancer cells. Unlike conventional kinase inhibitors that solely suppress catalytic activity, 11l induced G2/M phase arrest and downregulated CDK1, cyclin B1, and the replication initiation factor CDC45. Further investigation revealed that 11l induces severe DNA replication stress, subsequently activating the p53 signaling pathway to trigger apoptosis. This mechanism was recapitulated in CDC45 knockdown models. In vivo efficacy evaluation demonstrated that 30 mg/kg 11l achieved a tumor growth inhibition (TGI) rate of 56.4%, without inducing significant body weight loss or observable organ toxicity. Collectively, these findings identify 11l as a safe CDK1 inhibitor with a distinct mechanism of action, supporting its potential as a promising therapeutic strategy for cancer treatment.

Arenobufagin (ARBU), a steroid compound extracted from the venom of Bufo gargarizans, exhibits multi-target pharmacological activities, yet its role in regulating ferroptosis in gastric cancer stem cells (GCSCs) remains unclear. This study systematically evaluated the antitumor effects and mechanisms of ARBU using in vitro sphere culture, organoid models, and xenografts. ARBU inhibited GCSC proliferation and sphere formation in a concentration-dependent manner, reduced EdU incorporation and SOX2 expression in organoids, and markedly suppressed tumor growth in vivo while downregulating SOX2 and Nanog, with favorable biosafety. Mechanistically, ARBU induced ferroptosis, evidenced by elevated MDA, ROS, and Fe2+, decreased GSH and SOD, mitochondrial damage, COX2 upregulation, and GPX4/SLC7A11 downregulation. RNA-seq and functional studies further revealed that HCAR1 critically regulates GCSC self-renewal and antioxidant defense, and ARBU promoted ferroptosis via HCAR1 suppression. Collectively, these results demonstrate that ARBU inhibits GCSC proliferation and stemness by inducing ferroptosis through downregulation of the HCAR1 pathway, highlighting its potential as a therapeutic candidate for gastric cancer.

Red-fleshed plumcots are prone to flesh discoloration during cold storage, reducing marketability. We evaluated a mobile nitric oxide-integrated hypobaric packaging (NO+HYP) system. Control fruit exhibited fading red color, increased discoloration index and decreased total anthocyanin content. In contrast, pretreatment with 20 μL L-1 NO combined with 50 kPa hypobaric packaging delayed discoloration and preserved anthocyanins. Targeted metabolomics showed that NO+HYP broadly maintained the levels of major anthocyanin monomers without altering their relative proportions. Biochemical analyses indicated that NO+HYP concurrently activated anthocyanin biosynthesis enzymes and inhibited degradation enzymes. Moreover, NO+HYP alleviated oxidative stress by boosting antioxidant enzyme activities and the ascorbate–glutathione cycle, reducing ROS accumulation and non-enzymatic anthocyanin oxidation. It also suppressed cell wall-degrading enzymes, lowered MDA content and electrolyte leakage, thereby reinforcing pectin integrity and cellular compartmentalization to prevent pigment degradation. This synergistic NO+HYP strategy offers a promising engineering intervention to mitigate pigment deterioration and extend fresh produce shelf life.

Green adhesives are gradually replacing traditional synthetic adhesives due to their eco-friendly properties and sustainability. However, the industrial applications of green adhesives are limited by their lower strength and durability, longer curing time, higher cost compared to traditional synthetic adhesives. Here, inspired by the coacervation process of sandcastle worms and Dopa chemistry of mussels, we show green adhesives with combined high instant adhesive strength and low cost by enhancing the interaction of γ-polyglutamic acid (γ-PGA)/ε-poly-L-lysine (ε-PLL) coacervates through the introduction of tannic acid (TA). The preparation method is green, simple and fast, adhering to green chemistry principles. Specifically, taking advantage of variable non-covalent interactions between γ-PGA, ε-PLL and TA, the synergistical enhancement of cohesion and adhesion leads to the excellent adhesive strength of coacervates. As a result, the adhesive exhibits maximum adhesive strength of 18.21 MPa and instant adhesive strength of 1.54 MPa, surpassing most currently reported adhesives based on non-covalent bonds. Notably, the multiple non-covalent interactions endow the adhesive with reversible debonding-on-demand and recyclability, retaining its original adhesive strength even after 10 cycles. The feasibility to simultaneously overcome the major bottlenecks for green adhesives provides new insights for developing other sustainable green materials.

Background: Hydroxysafflor Yellow A (HSYA), the major bioactive component from Carthamus tinctorius L., exerts significant protective effects against myocardial ischemia-reperfusion injury (MIRI). Mitophagy is pivotal in the pathological process of MIRI, yet the specific molecular mechanism underlying HSYA-mediated mitophagy regulation remains unclear. Objective: This study aimed to investigate the association between HSYA treatment and mitochondrial autophagy in murine MIRI and to explore the potential mechanistic role of the SIRT1-FOXO3-BNIP3 signaling pathway using functional loss-of-function and rescue experiments. These findings may provide preliminary evidence supporting the clinical translational potential in MIRI therapy. Methods: Mouse myocardial ischemia-reperfusion injury (MIRI) model and oxygen-glucose deprivation/reoxygenation (OGD/R)-induced AC16 cardiomyocyte injury models were established. Metabolomics, molecular docking, and surface plasmon resonance (SPR) techniques were combined to screen the potential targets of HSYA. The SIRT1 inhibitor EX527 and SIRT1 siRNA were used to verify the underlying mechanism. Cardiac function, myocardial infarct size, mitochondrial function, the expression of autophagy-related proteins, and protein-protein interaction were detected and analyzed. Results: Compared with the MIRI group, HSYA significantly improved cardiac function in mice, as evidenced by increased left ventricular ejection fraction (LVEF) and left ventricular fractional shortening (LVFS) (p < 0.01), attenuated ST-segment elevation, and improved myocardial perfusion. HSYA also markedly reduced myocardial infarct size (p < 0.01) and serum levels of CK-MB, LDH, and cTnI (all p < 0.01) and ameliorated myocardial histopathological damage and mitochondrial ultrastructural integrity. Mechanistic studies revealed that HSYA significantly upregulated the expression of SIRT1, FOXO3, BNIP3, Beclin-1, and the LC3II/I ratio while downregulating p62 expression (p < 0.01), consistent with enhanced mitophagy-related activity. Furthermore, these protective effects were markedly attenuated upon SIRT1 inhibition or siRNA-mediated silencing, whereas HSYA intervention partially reversed these alterations. Additionally, co-immunoprecipitation (Co-IP) and pull-down assays demonstrated that HSYA promoted protein-protein interactions between SIRT1-FOXO3, FOXO3-BNIP3, and BNIP3-LC3B. Conclusions: These findings highlight that HSYA is associated with improved cardiac function, enhanced mitophagy-related activity, and upregulated SIRT1-FOXO3-BNIP3 signaling, providing robust experimental evidence for its clinical translational application in MIRI treatment.

Molecular glue degraders (MGDs) have emerged as a transformative modality in the field of targeted protein degradation (TPD), enabling the selective elimination of disease-relevant proteins, including those traditionally considered undruggable. Unlike bifunctional proteolysis-targeting chimeras (PROTACs), MGDs operate through monovalent architectures that induce protein–protein interactions (PPIs) between E3 ligases and neosubstrates, offering advantages in chemical simplicity, cell permeability, and target scope. However, MGD discovery remains serendipitously, and a translational framework that links rational design to predictable selectivity and tissue exposure is still lacking. In this review, we present an integrated framework for advancing next-generation MGDs through three critical dimensions: rational design, specificity optimization, and delivery systems. First, we examined cutting-edge strategies in MGD design, including covalent handle-based reprogramming, PPI-driven stabilization, and multi-site, multi-functional constructs. Second, we explored structure-guided engineering and chemoinformatic models, such as cereblon degron motifs, zone-based design and multiparameter optimization, to improve neosubstrate selectivity while minimizing off-target liabilities. Third, we summarized delivery platforms, including antibody‒drug conjugates, nanoparticle-enabled systems, and folate-mediated targeting, which are primarily intended to improve tissue selectivity and targeted distribution, thereby promoting local tissue accumulation. Finally, we discussed emerging opportunities at the intersection of artificial intelligence, structural biology, and systems pharmacology for accelerating MGD discovery and clinical translation. Collectively, these interdisciplinary insights underscore the therapeutic promise of MGDs and lay the groundwork for their next-generation evolution in precision medicine.

The management of breast cancer remains clinically intractable, driven by its highly invasive behavior and limited susceptibility to conventional treatments. In this study, we engineered an innovative cyclodextrinporphyrin co-assembled nanoplatform (CT NPs) to enable multimodal breast cancer therapy. By successfully encapsulating camptothecin (CPT) within this nanocarrier, the system (CTC NPs) achieved synergistic chemo-phototherapeutic efficacy through dual-modality action. The highly biocompatible cyclodextrin carrier significantly improved the physicochemical characteristics of CPT. In vivo studies revealed that CTC NPs effectively evaded clearance by the reticuloendothelial system, overcame the defect of premature drug leakage, and exhibited superior tumor targeting and infiltration capabilities. Under near-infrared (NIR) laser irradiation, CTC NPs can simultaneously induce localized hyperthermia and produce reactive oxygen species (ROS), thereby achieving efficient tumor ablation. In 4T1 tumor-bearing mice, CTC NPs exhibited targeted, safe, and highly potent anti-tumor efficacy, significantly suppressing both primary tumor progression (tumor suppression rate > 95 %) and metastatic dissemination. In summary, this integrated nanoplatform establishes a novel theranostic paradigm for synergistic chemo-phototherapy against triple-negative breast cancer (TNBC), achieving precise tumor ablation through NIR-triggered drug release and real-time imaging guidance. (c) 2026 Published by Elsevier B.V. on behalf of Chinese Chemical Society and Institute of Materia Medica, Chinese Academy of Medical Sciences.

Epigenetic dysregulation plays a critical role in tumorigenesis and cancer progression. The development of isoform-selective histone deacetylase (HDAC) inhibitors has emerged as a promising strategy in cancer therapy. In this study, based on our previously identified hit compounds, a series of novel N-arylamide-quinoline derivatives were rationally designed and synthesized as HDAC isoform-selective inhibitors with improved efficacy and reduced toxicity. Among them, 6b exhibited potent inhibitory activity against HDAC1, 2, 3, and 10, while showing no activity against HDAC4-9, a selectivity profile further supported by molecular docking and molecular dynamics simulation. 6b demonstrated significant antiproliferative effects against HL-60, CCRF-CEM, and HepG2 cancer cells. In vitro toxicity assays revealed a high selectivity index for this compound, markedly superior to that of the parent compounds. Mechanistic studies showed that it induced hyperacetylation of histone H3 in a concentration-dependent manner, downregulated Rb protein and caspase-8 precursor, and modulated the expression of BAX, BAK, and BCL-2, leading to extrinsic apoptosis and G0/G1 phase cell cycle arrest. In vivo, 6b exhibited potent antitumor activity with no apparent toxicity following both intragastric administration and intraperitoneal injection. Notably, intraperitoneal delivery resulted in enhanced efficacy. Pharmacokinetic studies further characterized the in vivo behavior of this compound via both routes. Overall, hit compound 6b displays favorable biological properties and represents a promising candidate for further anticancer drug development, with subsequent studies focusing on the optimization of its drug-like properties.

Background Ulcerative colitis (UC) is a chronic inflammatory disease that seriously endangers human health. Shaoyao Decoction (SYD) has been widely used in clinical practice to treat UC; however, its active components and molecular mechanisms remain unclear. Objective To elucidate the pharmacological mechanisms by which SYD and its active constituents ameliorate UC using single-cell RNA sequencing (scRNA-seq). Methods We first assessed the therapeutic efficacy of SYD in a dextran sulfate sodium (DSS)-induced UC mouse model. We then applied scRNA-seq to identify the potential cell populations and molecular pathways involved in SYD-mediated protection. Guided by these findings, we verified the inferred mechanisms in vivo and screened potential active compounds from SYD. We further examined their inhibitory effects on formyl-methionyl-leucyl-phenylalanine (fMLP)-induced neutrophil migration and their protective effects on DSS-induced Caco-2 cell injury in vitro. Finally, we validated the underlying mechanisms using inhibitors, agonists, gene silencing, and overexpression approaches in vitro. Results SYD markedly alleviated weight loss, reduced disease activity index (DAI) scores, and mitigated colon shortening and histopathological damage in mice with UC. The scRNA-seq analysis revealed alterations in neutrophils and intestinal epithelial cells. Functional pathway analysis indicated that neutrophil migration, epithelial tight junction regulation, and apoptosis were key processes modulated by SYD. In vivo, SYD decreased neutrophil infiltration and downregulated the expression of RAC1, RAC2, S100A8, and S100A9. Concurrently, SYD upregulated the tight junction proteins TJP1 and OCLN and suppressed epithelial apoptosis. In vitro, Ferulic acid emerged as the potential compound responsible for suppressing neutrophil migration. RAC1 inhibition and Ferulic acid treatment produced comparable suppressive effects on neutrophil migration, whereas RAC1 activation effectively antagonized the inhibitory effect of Ferulic acid. Chrysin-7-O-glucuronide was identified as the potential component enhancing tight junction integrity and suppressing apoptosis in intestinal epithelial cells; silencing KLF4 eliminated these protective effects of Chrysin-7-O-glucuronide. Conversely, KLF4 overexpression exerted protective effects comparable to those of Chrysin-7-O-glucuronide treatment. Conclusion SYD exerts potent therapeutic effects against UC. Mechanistically, Ferulic acid in SYD suppresses RAC1-mediated neutrophil migration, while Chrysin-7-O-glucuronide enhances tight junction integrity and suppresses apoptosis in intestinal epithelial cells through a KLF4-dependent mechanism.

Cerebral ischemia-reperfusion (CIR) injury disrupts the blood-brain barrier (BBB), leading to exacerbated brain damage. Muscone, the main active component of musk, has been reported to exert neuroprotective effects, but its mechanism in protecting BBB integrity remains unclear. In a mouse model of middle cerebral artery occlusion and reperfusion, muscone treatment significantly reduced Zea Longa scores, cerebral infarct volume, and increased the proportion of normal neurons. Laser speckle contrast imaging and small animal super-resolution ultrasound imaging showed that muscone promoted blood flow restoration in the ischemic hemisphere. Muscone also inhibited apoptosis of brain microvascular endothelial cells (BMECs), as evidenced by a decreased proportion of TUNEL⁺/CD31⁺ cells, reduced expression of pro-apoptotic proteins BAX and Cleaved-Caspase-3, and increased expression of anti-apoptotic protein Bcl-2. Furthermore, muscone attenuated the degradation of tight junction proteins (ZO-1, Occludin, Claudin-5) and reduced Evans blue leakage, indicating preserved BBB integrity. Mechanistically, muscone increased the phosphorylation of PKA and RHOA, while decreasing p-MLC expression in the ischemic hemisphere. And the PKA inhibitor H-89 reduced the protective effects of muscone on BMEC apoptosis, tight junction degradation, and Evans blue leakage, ultimately leading to increased Zea Longa scores, infarct volume, and neuronal damage. These findings demonstrate that muscone may exert a protective effect against CIR-induced BBB injury by inhibiting BMEC apoptosis and tight junction degradation through activation of the PKA/RHOA/MLC pathway. This study provides new insights into the mechanism of muscone and supports its potential application in ischemic stroke therapy.

Background: Plasma fibrinogen (FIB) levels exhibit a significant elevation during the acute phase of ischemic stroke (IS), and their dynamic fluctuations serve as important biomarkers for stroke onset, disease progression, and long-term prognosis. Tong-Qiao-Huo-Xue Decoction (TQHXD) is highly effective in treating blood stasis syndromes affecting the head and face. Nevertheless, the association between TQHXD and FIB in the underlying mechanism of treating IS warrants further investigation. Methods: Proteomics analysis predicted the potential therapeutic targets of TQHXD for IS. An in vivo model of middle cerebral artery occlusion followed by reperfusion (MCAO/R) was created in mice. To explore the interaction between FIB and NLRP3, as well as to verify the particular healing outcomes of TQHXD. Results: An increased blood-brain barrier (BBB) permeability was observed after MCAO/R, accompanied by substantial accumulation of FIB in the brain. In vivo experiments demonstrated that FIB triggered the activation of the NLRP3 inflammasome in microglia. Proteomic analysis revealed a significant increase in FIB levels following model induction, which were markedly reduced after treatment with TQHXD; KEGG pathway enrichment analysis indicated that these changes were primarily associated with the NOD-like receptor signaling pathway. Laser speckle contrast imaging showed that TQHXD treatment significantly improved cerebral blood flow and attenuated brain injury in mice. Fluorescence imaging, ELISA, and Western blotting results collectively demonstrated that TQHXD effectively reduced FIB accumulation and suppressed NLRP3 inflammasome activation. MD and pull-down experiments further demonstrated a strong interaction strength between FIB and NLRP3. Conclusions: FIB accumulates in the ischemic penumbra following CIRI, while TQHXD can effectively down-regulate FIB expression and inhibit NLRP3 inflammasome activation to mitigate CIRI. These findings provide a novel theoretical foundation and treatment direction for stroke management in clinical settings.

Objective Microgravity-induced neural dysfunction poses a critical risk during long-term spaceflight. However, the intrinsic mechanosensing mechanisms by which neural stem cells (NSCs) decode gravitational loss into biochemical responses and effective multi-target countermeasures remain elusive. This study aims to clarify the protective mechanism of ginsenoside Rg1 (Rg1) on simulated microgravity (SMG)-damaged C17.2 mouse neural stem cells (NSCs) by targeting vimentin serine 56 (Ser56) phosphorylation and the mitogen-activated protein kinase (MAPK)/protein kinase B (AKT) signaling network, and verify its “multi-pathway synergy” characteristic guided by traditional Chinese medicine (TCM) theory. Methods An SMG environment was established using a random positioning machine (RPM). The safe and effective concentration of Rg1 (40 μmol/L) was screened using the cell counting kit-8 (CCK-8) assay. Mitochondrial function and cytoskeletal structure were evaluated via transmission electron microscopy and F-actin staining. Flow cytometry, Western blotting, quantitative real-time polymerase chain reaction, combined with global proteomics, phosphoproteomics, and extracellular signal-regulated kinase 1/2 (ERK1/2) inhibitor experiments, were employed to dissect the action mechanism of Rg1. Results First, SMG inhibited NSC differentiation, disrupted cytoskeletal structure, and induced apoptosis, with the core mechanism being the impairment of cytoskeletal architecture and its dynamic phosphorylation network. Second, 40 μmol/L Rg1 restored F-actin continuity, rescued neurodifferentiation function, and inhibited apoptosis; these effects relied on the activation of the MAPK/AKT kinase network and the specific rescue of vimentin Ser56 phosphorylation. Third, Rg1 regulated the kinase-intermediate filament (IF) axis through “ERK1/2-dependent and independent” dual pathways rather than single-target intervention. Notably, its regulatory effect directly acted on post-translational modification (phosphorylation) instead of merely altering protein abundance. Conclusion Rg1 restores cytoskeletal integrity and neurogenesis in C17.2 NSCs under SMG by regulating the kinase-IF axis. This modern molecular evidence bridges TCM theory with mechanobiology, confirming Rg1 as a promising TCM candidate for protecting astronaut brain health. It exemplifies the “holistic regulation” of TCM and provides a paradigm for the application of TCM in space medicine.

BACKGROUND:Traditionally, wild jujube (Ziziphus jujuba Mill. var. spinosa (Bunge) Hu ex H. F. Chou) has been used to nourish the heart, calm the spirit, and arrest spontaneous sweating. Modern research confirms its broad pharmacological activities, including antioxidant, anti-inflammatory, neuroprotective, and cognitive-enhancing effects. This study aims to isolate and characterize the structure of jujube polysaccharides and evaluate their protective effects against oxidative stress damage in neural stem cells (NSCs). METHODS:We successfully isolated and purified a novel pectin polysaccharide (ZJP-2) from wild jujube. Its structure was characterized in detail using high-performance liquid chromatography coupled with multi-angle laser light scattering and refractive index detection (HPLC-MALS-RI), high-performance anion exchange chromatography (HPAEC), gas chromatography-mass spectrometry (GC-MS), and nuclear magnetic resonance (NMR) spectroscopy. RESULTS:Structural analysis revealed that ZJP-2 is a pectin heteropolysaccharide with a molecular weight of approximately 67.93 kDa. Its monosaccharide composition primarily includes galac-turonic acid (GalA), arabinose (Ara), rhamnose (Rha), galactose (Gal), and glucose (Glc). The backbone consists of α-GalA and rhamnose-galacturonic acid-I (RG-I) domains linked by (1→4)-glycosidic bonds. NMR spectroscopy further confirmed its glycosidic bond types. In activity assessment, our study demonstrated that ZJP-2 significantly alleviated DMNQ-induced oxidative stress damage in C17.2 neural stem cells. Its protective effect was achieved by reducing intracellular reactive oxygen species (ROS) levels and upregulating the mRNA expression of antioxidant genes associated with the signaling axis (p < 0.05). Moreover, ZJP-2 suppressed DMNQ-induced overexpression of Nestin and NeuN (p < 0.05), contributing to the maintenance of NSCs' undifferentiated state and functional homeostasis. CONCLUSIONS:In conclusion, ZJP-2 possesses distinct structural characteristics and significant neuroprotective potential, supporting its development as a natural functional food or dietary supplement for preventing oxidative stress-related neural damage.

BACKGROUND:Tong-Qiao-Huo-Xue Decoction (TQHXD), a classical Chinese medicine formula, is widely used to enhance blood circulation, remove stasis, and improve outcomes in ischemic stroke. Its mechanisms in cerebral microarteriogenesis remain unclear. PURPOSE:This study investigated the role of TQHXD in promoting cerebral microarteriogenesis after ischemic stroke and explored S1P-mediated communication between brain microvascular endothelial cells (BMECs) and brain microvascular smooth muscle cells (BMSMCs). STUDY DESIGN:An integrated approach combining metabolomics, molecular docking, MCAO/R rat models, and in vitro BMSMC assays was employed to elucidate TQHXD neurovascular protective mechanisms. METHODS:Stroke-related targets were predicted via metabolomics, and UHPLC-MS/MS identified brain-penetrating active components. Molecular docking evaluated their binding affinity to S1P1. MCAO/R rats were assessed for neurological function, neuronal apoptosis, and cerebral microvascular morphometry. In vitro, BMSMC viability, proliferation, migration, phenotypic switching, and angiogenic factor expression were analyzed. Western blotting, co-immunoprecipitation, and pull-down assays validated key signaling pathways and protein-protein interactions. RESULTS:TQHXD improved neurological deficits, reduced cortical neuronal apoptosis, and increased microvascular density, length, and perfusion. CSF-detectable components (muscone, amygdalin, ligustilide, paeoniflorin, and hydroxysafflor yellow A) exhibited high S1P1 affinity. In vitro, TQHXD enhanced BMSMC viability, proliferation, migration, and phenotypic switching, activated S1P1/RAS/RAF/MEK/ERK signaling, and upregulated angiogenic and neurotrophic factors (PDGF, VEGF, bFGF, BDNF). Co-immunoprecipitation and pull-down assays confirmed specific protein-protein interactions within the S1P-mediated cascade. CONCLUSION:TQHXD confers neurovascular protection by activating S1P-mediated S1P1/RAS/RAF/MEK/ERK signaling, promoting cerebral microarteriogenesis and collateral circulation restoration, providing mechanistic evidence supporting its clinical application in ischemic stroke.

OBJECTIVES:To examine the protective effects of cerebrospinal fluid from Tongqiao Huoxue Decoction-treated rats (TQHXD-CSF) against oxygen and glucose deprivation and reoxygenation (OGD/R)-induced injury in murine BV-2 microglial cells and co-cultured HT22 cells. METHODS:In a Transwell co-culture system of BV-2 and HT22 cells, OGD/R+fibrinogen (FIB) injury was induced in BV-2 cells followed by treatment with TQHXD-CSF intervention, and HT22 cells in the lower chamber were cultured under normal conditions. The cells were observed for changes in cell morphology, viability, intracellular ROS level, apoptosis, M1/M2 polarization, and FIB-containing extracellular vesicles (EVs). The cellular expressions of NLRP3, ASC, caspase-1, GSDMD, IL-1β, and IL-18 were quantified using Western blotting, and FIB-NLRP3 binding was confirmed by pull-down assay. RESULTS:OGD/R+FIB injury caused polarization of BV-2 cells to the pro-inflammatory M1 phenotype, increased CD86 expression and release of FIB-containing EVs, and activated the NLRP3 inflammasome. The co-cultured HT22 cells showed reduced cell viability, elevated ROS, and increased cells apoptosis. Treatment with TQHXD-CSF promoted M2 polarization in BV-2 cells, upregulated CD206 expression, suppressed FIB+ EVs secretion, and inhibited NLRP3 inflammasome activation in HT22 cells, which showed significantly lowered expressions of NLRP3, ASC, caspase-1, GSDMD, IL-1β, and IL-18 proteins and hence reduced inflammatory injury and cell apoptosis. Pull-down assay confirmed direct FIB and NLRP3 binding. CONCLUSIONS:In the co-culture system of BV-2 cells and HT22 cells, TQHXD-CSF treatment protects HT22 cells against OGD/R+FIB-induced injury by inhibiting FIB-containing EVs release from BV-2 cells and suppressing NLRP3 inflammasome activation.

Acute liver injury (ALI) is a severe public health problem closely associated with oxidative stress, inflammation, and hepatocyte injury, leading to high mortality. Fucoxanthin (Fx), a marine carotenoid found in brown seaweeds, has various beneficial effects against multiple diseases. However, the potential role of Fx on ALI remains unclear. This study aims to explore the pharmacological potential of Fx in lipopolysaccharide (LPS)/D-galactosamine (D-Gal)-induced ALI. The therapeutic effect of Fx on ALI was primarily evaluated using a mouse model induced by LPS/D-Gal, focusing on pathological changes, oxidative stress, inflammation, and pyroptosis. Additionally, the effects of Fx on cell pyroptosis and its molecular mechanisms were explored in an in vitro pyroptosis model established by inducing macrophages with LPS/Nigericin. Fx significantly alleviated the LPS/D-Gal-induced histopathological progression and hepatocyte apoptosis, reducing plasma levels of ALT, AST, and LDH. It also obviously decreased hepatic MDA levels while increasing antioxidant enzyme activities and GSH concentration compared to LPS/D-Gal-treated mice. These antioxidant effects were linked to the upregulation of hepatic Nrf-2, HO-1, and GCLC expression. Furthermore, Fx treatment alleviated macrophage accumulation and downregulated the expression of pro-inflammatory factors in the liver. Importantly, Fx administration suppressed NLRP3 inflammasome-dependent canonical pyroptosis both in LPS/D-Gal-treated mice and LPS/Nigericin-stimulated macrophages, potentially mediated by the suppression of MAPKs and NF-κB pathways. These findings suggest that Fx could be an effective strategy to prevent ALI, particularly in cases associated with NLRP3 inflammasome-mediated pyroptosis.

Microgravity during long-duration spaceflights promotes oxidative damage, impairs mitochondrial efficiency, and induces neurodegeneration, highlighting the need for safe countermeasures. Gastrodia elata (Tianma) is a traditional medicinal herb with known antioxidant and neuroprotective properties that has not yet been examined under microgravity-analog conditions. The main aim of this study was to evaluate whether Tianma mitigates oxidative, mitochondrial, and neurobehavioral deficits induced by simulated microgravity (SM) in Caenorhabditis elegans (C. elegans). N2, TJ356 (DAF-16::GFP), and BZ555 (dat-1p::GFP) worms were treated with Tianma extract or vehicle for 72 h and exposed to simulated microgravity for 24 h thereafter. We quantified multiple functional and molecular endpoints related to organismal health, redox balance, mitochondrial function, dopaminergic neuron integrity, and stress resistance to determine the mitigative effect of Tianma on SM. SM significantly disrupts organismal and cellular homeostasis, as evidenced by reduced locomotion, pharyngeal pumping, associative learning and memory with butanone, dopamine-dependent basal slowing, and reduced lifespan. These functional deficits were accompanied by elevated oxidative damage, diminished antioxidant enzyme activities, mitochondrial depolarization and ATP depletion, enhanced DAF-16 nuclear translocation, upregulation of antioxidant, heat-shock, and mitochondrial stress-response genes, and progressive dopaminergic neuron loss. Tianma treatment restored behavioral performance, extended lifespan, normalized oxidative markers and enzyme activities, preserved mitochondrial function, limited DAF-16 nuclear accumulation, attenuated stress-gene induction, protected dopaminergic neurons, and enhanced stress resistance. These findings indicate that Tianma provides integrated antioxidant, mitochondrial, and neuroprotective effects under simulated microgravity and merit further evaluation as a potential countermeasure against spaceflight-associated oxidative and neurodegenerative stress.