Hemorrhagic transformation (HT), characterized by secondary hemorrhage into the brain tissue following cerebral infarction, is a serious complication of ischemic stroke. As an independent risk factor, hyperglycemia exacerbates stroke by inducing HT, thereby increasing mortality and disability and posing an urgent clinical need. Neutrophil extracellular traps (NETs), a mediator of inflammation, are emerging as a therapeutic target after stroke. Our previous study demonstrated that NETs aggravate hyperglycemia-induced HT, however, the regulatory mechanisms of hyperglycemia in NETs formation are poorly defined. This study aimed to elucidate the mechanisms of STAT3 participating in hyperglycemia-induced HT by regulating NETs information. Network pharmacology results firstly suggested that hyperglycemic stroke-induced HT may be related to STAT3 and the regulation of NETs. Neutrophil counts, NLR (%) and NETs levels were found positively correlated with blood glucose in stroke patients. Further experiment suggested that STAT3 was activated both in high-glucose neutrophils and hyperglycemic MCAO rats. Both NETs inhibition with DNase I and STAT3 inhibition with Stattic could improve HT and BBB integrity in hyperglycemic MCAO rats, combination administration demonstrated greater efficacy. Inhibition of STAT3 alleviated thromboinflammation in hyperglycemia-induced HT rats by suppressing NETs formation and decreased levels of inflammatory factors such as IL-8 and TNF-α, and thrombosis-related factors such as vWF, TAT and Fibrinogen. This study established evidence that hyperglycemia exacerbates HT through STAT3 activation which promotes NETs formation, suggesting STAT3 as a novel target for hyperglycemia-induced HT.

Abstract Xing-Qi-Tong-Qiao Decoction (XQTQ) is a traditional Chinese prescription that embodies the homology of medicine and food. It is recognized for its effects in alleviating stasis, dispersing phlegm and nodules, activating blood circulation, relieving pain. However, its efficacy and mechanisms in alleviating paclitaxel-induced peripheral neuropathic pain (PIPNP) remain unclear. In this study, we investigate the analgesic effects of XQTQ and explore the potential mechanism using plasma metabolomics. The characteristic chromatogram of XQTQ was established using HPLC. A total of 36 SD rats were divided into 6 groups: control, model, positive drug, and three XQTQ combined groups. All rats, except those in control group, were induced with diseases features via intraperitoneal administration of paclitaxel. The pregabalin and XQTQ combined groups were administered orally once a day for 14 days. The paw withdrawal threshold (PWT) and paw withdrawal latency (PWL) were measured. Rat plasma was collected for the detection of pain cytokines and the analysis of endogenous metabolite. Rats in the XQTQ combined groups exhibited a significant increase in PWT and PWL values, alongside a decrease in the levels of tumor necrosis factor-alpha (TNF-α, 14.82%), interleukin-6 (IL-6, 13.69%), interleukin-1 beta (IL-1β, 19.34%), nerve growth factor (NGF, 11.40%), prostaglandin E2 (PGE 2, 12.74%), and 5-hydroxytryptamine (5-HT, 13.51%) compared to the model group (P < 0.01). Plasma metabolomics revealed 12 potential biomarkers associated with the model rats, which were significantly reversed by XQTQ. The main pathways involved included glycerophospholipid metabolism, linoleic acid (LA) metabolism, and alpha-linolenic acid (ALA) metabolism. XQTQ demonstrated significant analgesic effects and improved metabolism in the context of PIPNP. This research provides a reliable theoretical basis and novel research directions for clinical applications.

Psychobiotics are beneficial microorganisms or their metabolites that modulate the gut microbiome to enhance mental health. However, the oral delivery of live bacteria faces several challenges, including inactivation by gastric acid, low intestinal colonization rates, poor targeting, and considerable interindividual variability in efficacy, which impede their clinical translation. This study focuses on artificial intelligence (AI)-driven psychobiotic nanodelivery systems. Initially, it summarizes common psychobiotics and their mechanisms targeting the gut-brain axis for treating psychiatric disorders. It then outlines the main types of probiotic delivery systems and their applications in mental health treatment. Furthermore, the significant role of artificial intelligence in strain selection and optimization, personalized treatment design, and clinical outcome evaluation is explored. Finally, the study anticipates the prospects and challenges of integrating artificial intelligence with nanodelivery systems to enhance the therapeutic efficacy of psychobiotics, aiming to provide valuable insights for related research and clinical applications.

Probiotics are a category of beneficial microorganisms that contribute to human health. In recent years, there has been considerable interest in the potential of probiotics as dietary supplements to regulate host mental health. However, research on the systematic functions of specific strains remains limited. In the present work, the chronic unpredictable mild stress (CUMS) rat model was constructed, and it was found that the abundance of Lactobacillus Johnsonii in intestinal tract was significantly reduced. Conversely, exogenous supplementation of Lactobacillus johnsonii significantly improved mood-related behaviors of the CUMS rats. Furthermore, analysis of serum and colonic contents metabolomics revealed that the levels of key metabolites of the tryptophan metabolic pathway (tryptophan, indole-3-acetic acid, and 5-HT) were abnormally reduced in depressed model rats. The changes in the concentrations of these metabolites were significantly and positively related to the degree of mood-related behaviors. Supplementation with Lactobacillus johnsonii restored the levels of the tryptophan metabolites described above. In addition, further studies demonstrated that Lactobacillus johnsonii intervention could effectively improve intestinal barrier damage by upregulating the ZO-1 and Claudin1 tight junction proteins, and inhibit intestinal inflammatory responses by suppressing the expression of TNF-alpha, IL-1 beta, IL-6, and IL-17A inflammatory factors. The present work reveals the potential mechanism of Lactobacillus johnsonii to alleviate chronic stress-induced mood-related behaviors by modulating tryptophan metabolism, improving intestinal barrier function, and inhibiting inflammatory pathways. In summary, the result demonstrates the significant role that Lactobacillus johnsonii could play as a functional psychotropic probiotic in ameliorating mood-related behaviors.

Lacidipine (LCDP) is a dihydropyridine calcium channel blocker and is considered a first-line agent for the clinical treatment of hypertension. In this work, four novel LCDP solvates, including dimethylacetamide solvate (SDMA), dimethylformamide solvate (SDMF), 1,4-dioxane solvate (SDIOX), and dimethyl sulfoxide solvate (SDMSO) were obtained by systematic crystallization screening. The four LCDP solvates were characterized by various analytical techniques and the crystal structures of these solvates were determined by single crystal X-ray diffraction for the first time. Furthermore, the phase transformation relationships of the aforementioned forms were investigated. The results revealed that LCDP tended to form solvates with solvent molecules containing only hydrogen bond acceptors via N-H & sdot;& sdot;& sdot;O interactions. In this motif, the secondary amine group on the dihydropyridine ring of LCDP acted as the hydrogen bond donor, while oxygen atoms from the solvent molecules served as acceptors. These intermolecular hydrogen bonds played a critical role in solvate formation.

Melanoma is an aggressive malignant tumor with an increasing incidence worldwide. Ganglioside GD3 is recognized as a human melanoma-specific antigen and is highly expressed in melanoma tissues and cell lines. GD3 synthase (GD3s) is the rate-limiting enzyme for the synthesis of GD3. However, the function and mechanism of GD3s in melanoma remains poorly understood. The expression of GD3s was evaluated by an in silico analysis and detected by melanoma tissue microarray. Cell loss-of-function and gain-of-function, proliferation, colony formation, wound healing assay, transwell assay, tumor xenograft mouse model and a tail vein-injection mouse model were conducted to determine the functional role of GD3s in melanoma progression. Proteomic analysis and metabolomic analysis were used to identify downstream targets of GD3s. Quantification of triglyceride and cholesterol, nile red staining, Western blot, Real-time quantitative PCR (RT-qPCR) and Co-immunoprecipitation (Co-IP) assays were employed to validate the underlying mechanisms of GD3s which aggravate progression of melanoma. High-throughput screening method was used to identify the inhibitor of GD3s. Dysregulated high levels of GD3s were correlated with advanced clinical stages and poor prognoses of melanoma. GD3s promoted the proliferation and metastasis of melanoma by reprograming lipid metabolism. GD3s induced extensive lipid accumulation and elevated expression levels of lipogenic enzymes in melanoma cells by regulating AMPK/SREBP1 signaling pathway. ENMD-2076 L-(+)-Tartaric acid was considered to be a novel GD3s inhibitor and exhibited good anti-tumor activity both in vivo and in vitro. Mechanistically, consistent with shRNA-mediated silencing of GD3s, ENMD-2076 L-(+)-Tartaric acid inhibited de novo fatty acid synthesis through AMPK/SREBP1 pathway. The current findings uncovered a novel mechanism by which GD3s modulated aberrant lipid metabolism and promoted the progression of melanoma. ENMD-2076 L-(+)-Tartaric acid was firstly discovered as a new inhibitor of GD3s and had good anti-tumor effect both in vitro and in vivo.

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Ischemic stroke (IS) remains a devastating cerebrovascular disease associated with high morbidity, mortality, and disability, while effective restorative therapies remain limited, highlighting an urgent unmet clinical need. Post-stroke angiogenesis contributes to vascular remodeling, restoration of cerebral blood flow, and neurovascular regeneration, making it a promising therapeutic target for IS repair. Salvianolic acid A (SAA), a phenolic acid from Salvia miltiorrhiza (Danshen), protects vascular endothelial cells and enhances neurogenesis, but its role and mechanisms in post-stroke angiogenesis remain unclear. This study investigated the effects and molecular mechanisms of SAA on post-stroke angiogenesis in vitro and in vivo. In vivo, an autologous thrombus-induced rat stroke model was established by electrocoagulation to evaluate the effects of SAA on neurological recovery and angiogenesis. In vitro, hydrogen peroxide (H₂O₂)-induced oxidative damage in human brain microvascular endothelial cells (HBMECs) was used to investigate the protective effects of SAA on endothelial function. Network pharmacology, molecular docking, and surface plasmon resonance (SPR) identified targets, with mechanistic verification both in vitro and in vivo. In the rat stroke model, SAA improved neurological deficits and enhanced functional recovery in a dose- and time-dependent manner. It also increased peri-infarct microvessel density, promoted vascular maturation, and upregulated pro-angiogenic factors (VEGF-A, Ang-1) and tight junction proteins (TJPs) (ZO-1, Occludin, Claudin-5). In HBMECs, SAA alleviated oxidative stress, enhanced proliferation, migration, and tube formation. Network pharmacology and SPR confirmed AKT as a core target of SAA, further activating the Nrf2 signaling pathway. AKT inhibitor MK-2206 counteracted the pro-angiogenic effect of SAA. In conclusion, SAA promotes post-stroke angiogenesis by targeting AKT and activating Nrf2-mediated antioxidant signaling, thereby protecting vascular endothelial function and improving neurological recovery. These findings provide new insights into vascular repair after IS and highlight SAA as a potential therapeutic candidate for stroke recovery via modulating angiogenesis.

Objective Cerebral ischemia-reperfusion (I/R) injury contributes substantially to neuronal damage after stroke. Recent studies have demonstrated the efficacy of Glycyrrhizae radix et rhizoma in I/R injury, yet the underlying mechanism remains unclear. Therefore, the present study investigated whether isoliquiritin apioside (ISLA), a bioactive compound derived from it, protected against cerebral I/R injury. Methods Rats subjected to middle cerebral artery occlusion/reperfusion were treated with ISLA (5, 10, or 20 mg/kg). Neurological outcomes and infarct volume were evaluated, histopathological and apoptotic changes were assessed by H&E and TUNEL staining, oxidative stress was determined by MDA and SOD assays, the expression of proteins related to TXNIP/NLRP3 signaling, pyroptosis, and inflammation was examined by western blotting and immunofluorescence methods. Results ISLA significantly reduced infarct volume, improved neurological function, preserved cortical morphology, and decreased TUNEL-positive cells in the ischemic brain. ISLA also attenuated oxidative stress, as evidenced by reduced MDA levels and restored SOD activity. Mechanistically, ISLA downregulated TXNIP and NLRP3 expressions, suppressed cleaved caspase-1 and cleaved GSDMD production, and reduced IL-1β and IL-18 levels. Conclusion These findings indicated that ISLA alleviated cerebral I/R injury, at least in part, by reducing oxidative stress and inhibiting TXNIP/NLRP3-associated pyroptotic and inflammatory signaling.

Pharmaceutical solid form engineering through co-crystallization represents a transformative strategy for enhancing drug bioavailability and physicochemical properties, yet remains hindered by labor-intensive experimental screening and the notable absence of computational models capable of simultaneously classifying co-crystals, salts, and physical mixtures. Here, we present an intelligent multi-modal graph neural network (GNN) framework that synergistically integrates molecular graph representations, molecular fingerprints, and quantum chemical descriptors to enable automated three-class classification of pharmaceutical solid forms, addressing a critical gap in current computational pharmaceutical research. Our innovative feature fusion strategy combines topological molecular graphs with traditional molecular fingerprints and computationally-derived quantum chemical properties, creating a comprehensive multi-dimensional representation of pharmaceutical compounds. The developed integrated model achieves exceptional classification accuracy exceeding 98 % across all evaluation phases, demonstrating the superior performance of combined structural, topological, and electronic feature representations in complex multi-component pharmaceutical systems. Rigorous validation using three independent active pharmaceutical ingredient datasets, theophylline, ligustrazine, and piperazine, yields outstanding success rates, confirming the model's remarkable generalizability and practical applicability across diverse chemical spaces. Most significantly, our artificial intelligence (AI)-driven multi-modal predictions successfully guided the experimental synthesis of two novel co-crystalline phases: Emodin-ligustrazine and emodin-metformin, validating the predictive power of integrated molecular feature representations in real-world pharmaceutical development.

In recent years, immunotherapy has shown obvious advantages in treating cancers. The close interaction between cancer cells and immune cells in the tumor microenvironment (TME) underlies the progression of glioblastoma multiforme (GBM). However, there are no effective immune-related targets against GBM. Here, in silico analyses and experimental data showed that Interferon Gamma Inducible Protein 30 (IFI30), modulated by histone modifications both H3K4me3 and H3K27ac, was up-regulated in GBM and had a potential role in the antitumor immune responses. In vitro and in vivo experiments further revealed that IFI30 modulated the infiltration of tumor-associated macrophages (TAMs) and reduced the proportion of CD8+ T cells. Mechanistically, IFI30 induced PGE2 expression in GBM cells via the MAFF/PTGS2 pathway, and PGE2 bound to macrophage EP2/EP4, activating the downstream ERK1/2 and KLF4/STAT6 pathways, stimulating the infiltration of TAMs. Taken together, we characterized the role and mechanisms of IFI30 in the malignant progression of GBM by regulating TAMs, highlighting that IFI30 may benefit GBM patients as a therapeutic target.

Black tea is a kind of full-fermented tea with therapeutic potential for metabolic diseases. Ectopic lipid deposition (ELD) is an essential risk factor for organ injury in metabolic syndrome, especially in liver and kidney, for which effective interventions are lacking. Here, we explored whether black tea extract (BTE) improves fatty liver and fatty kidney, as well as identified the potential lipid biomarkers for ELD and lipid targets of BTE on the improvement of ELD. Transcriptome data from diet-induced obese mice were analyzed to confirm high-fat-diet feeding disturbs lipid metabolism in the liver and kidney. BTE prominently inhibited body weight gain, improved glucose metabolism, as well as reduced lipid droplet accumulation in the liver and kidney. Moreover, lipidomic profiling analyses identified 28 lipid classes in the liver while 29 in the kidney with that up-regulated glycerides and phosphatidylcholines, as well as down-regulated cardiolipin were the characteristic changes in ELD. BTE prominently reduced glycerides in ELD, thereby constituting the basis of its anti-ELD effect. Specifically, BTE displayed stronger effects on lowering cholesterol ester (CE) in fatty liver, while also affecting phospholipids and sphingolipids in fatty kidney. Ultimately, integrative analysis identified CE18:2 and triglyceride (TAG)56:4 (20:2) as the potential lipid biomarkers for BTE in improvement of ELD. BTE could be an effective food supplement for the prevention and treatment of ELD. Notably, CE18:2 and TAG56:4(20:2), as the potential lipid biomarkers, may facilitate the research and development of anti-ELD drug.

BACKGROUND:Pulmonary hypertension (PH) is characterized by pulmonary vascular remodeling, and Furin, a proprotein convertase critical for TGF-β1 activation, has shown promise as a novel therapeutic target for PH. However, its specific role in PH pathogenesis and potential inhibitors have remained unexplored. PURPOSE:This study aimed to investigate the role of Furin in PH-associated vascular remodeling and evaluate the therapeutic potential of the newly discovered Furin inhibitor, Licochalcone B (LicoB), in PH. METHODS:We assessed Furin expression levels in multiple PH animal models and evaluated its cellular localization using single-cell transcriptomics. Then the functional role of Furin was validated through overexpression and knockdown studies in vitro. The protective effect of Furin inhibitor, LicoB, was validated in pulmonary artery vascular cells, and finally its therapeutic effects and potential mechanisms were evaluated in PH animal models. RESULTS:Elevated Furin expression was consistently observed across diverse PH animal models. Furin overexpression promoted cell viability and migration, while its knockdown mitigated pathological features. The Furin inhibitor, LicoB, effectively suppressed aberrant pulmonary artery cell proliferation and migration, and restored mitochondrial membrane potential in vitro. Critically, in both murine and rat PH models, LicoB administration significantly ameliorated symptoms, exerted robust cardiopulmonary protection, and attenuated pulmonary vascular remodeling. Mechanistically, LicoB acted by inhibiting the Furin/TGF-β1 signaling axis. CONCLUSION:Our findings establish Furin as a key driver of PH development and demonstrate that LicoB suppresses vascular remodeling via the Furin/TGF-β1 pathway, positioning LicoB as a highly promising novel therapeutic candidate for PH by targeting Furin.

Background:Metabolic dysfunction-associated steatohepatitis (MASH) is the most prevalent chronic liver disease worldwide; however, few effective therapeutic options are available for MASH. Artemisia scoparia is a medicinal plant that has been widely utilized in traditional medicine to treat liver-related ailments. Nonetheless, the effects and underlying mechanisms of A. scoparia in the context of MASH remain poorly understood. Aim of the study:The objective of this research was to assess the protective effects and further mechanisms of A. scoparia extract (AS) on a MASH mice model. Methods:The protective effects of AS were evaluated both in vivo and in vitro, with the therapeutic efficacy of AS being characterized through the detection of biochemical markers, histological analysis, and Oil red O staining. To elucidate the underlying mechanisms and pharmacodynamic basis of AS, a comprehensive set of techniques were applied, including transcriptomics, metabolomics, Western blotting, and immunofluorescence staining. Results:AS reduced the blood lipid indices and inflammatory levels in the MASH mouse model and decreased lipid droplet accumulation in FFA-induced HepG2 cells. Transcriptomic and metabolomic analyses indicated that AS regulates 30 dysregulated genes (e.g., Gm15622, Pdia6, and Derl3) and controls 60 metabolic metabolites (e.g., heptadecanoic acid, 5b-cyprinol sulfate, and taurodeoxycholic acid) to ultimately affect core pathways involved in lipid metabolism and inflammation. Furthermore, AS was proven to exert a hepatoprotective effect by inhibiting inflammation and ferroptosis, along with weakening the advanced glycation end product-receptor for advanced glycation end products (AGE-RAGE) pathway and the Janus kinase-signal transducer and activator of transcription (JAK-STAT) pathway in vivo and in vitro. Conclusion:This study first elucidates the mechanism through which AS ameliorates MASH through integrated multi-omics analysis, providing experimental evidence for further development of natural therapeutic agents.

4-Methylumbelliferone (4-MU) is used clinically to treat conditions such as biliary dyskinesia, inflammation, autoimmune diseases, cancer, and other conditions. However, 4-MU suffers from poor solubility, low permeability, and suboptimal bioavailability, which limit its pharmacological activity and clinical application. Cocrystallization technology, as an effective method for improving physicochemical properties of drugs, offers a viable approach to address these issues with 4-MU. In this study, four cocrystal formers (CCFs) with druggable potential, namely nicotinamide (NAM), isonicotinamide (INA), theophylline (TP), and piperazine (PPZ), were selected for cocrystallization experiments with 4-MU, leading to the successful preparation of four novel cocrystals: 4-MU-NAM (1:1), 4-MU-INA (1:1), 4-MU-TP (1:1), and 4-MU-PPZ (2:1). Cocrystals were characterized using single-crystal X-ray diffraction (SXRD), powder X-ray diffraction (PXRD), differential scanning calorimetry (DSC), thermogravimetric analysis (TG), and infrared spectroscopy (IR). Solubility, permeability, and bioavailability were also evaluated for each cocrystal. In vitro results demonstrated that all four cocrystals improved the physicochemical properties of 4-MU, specifically: 4-MU-PPZ increased the water solubility of 4-MU to six times that of the parent drug, enhanced permeability to three times that of the parent drug, and significantly accelerated the drug's permeation rate. In vivo results indicated that 4-MU-TP effectively increased the bioavailability of 4-MU. Unlike traditional 4-MU structural modification methods, cocrystal engineering offers significant advantages of environmental friendliness and operational simplicity. This study effectively improved the key pharmaceutical properties of 4-MU through cocrystal technology, providing a new research direction and technical support for enhancing its druggability and promoting its application in more disease treatment fields.

In the pharmaceutical field, machine learning can play an important role in drug development, production and treatment. Co-crystallization techniques have shown promising potential to enhance the properties of active pharmaceutical ingredients (APIs) such as solubility, permeability, and bioavailability, all without altering their chemical structure. This approach opens new avenues for developing natural products into effective drugs, especially those previously challenging in formulation. Emodin, an anthraquinone-based natural product, is a notable example due to its diverse biological activities; however, its physicochemical limitations, such as poor solubility and easy sublimation, restricted its clinical application. While various methods have improved emodin's physicochemical properties, research on its bioavailability remains limited. In our study, we summarize cocrystals and salts produced through co-crystallization technology and identify piperazine as a favorable coformer. Conflicting conclusions from computational chemistry and molecular modeling method and machine learning method regarding the formation of an emodin-piperazine cocrystal or salt led us to experimentally validate these possibilities. Ultimately, we successfully obtained the emodin-piperazine cocrystal, which were characterized and evaluated by several in vitro methods and pharmacokinetic studies. In addition, experiments have shown that emodin has a certain therapeutic effect on sepsis, so we also evaluated emodin-piperazine biological activity in a sepsis model. The results demonstrate that co-crystallization significantly enhances emodin's solubility, permeability, and bioavailability. Pharmacodynamic studies indicate that the emodin-piperazine cocrystal improves sepsis symptoms and provides protective effects against liver and kidney damage associated with sepsis. This study offers renewed hope for natural products with broad biological activities yet hindered by physicochemical limitations by advancing co-crystallization as a viable development approach.

Colorectal cancer (CRC) incidence and mortality rates are steadily on the rise, which brings significant public health concern worldwide, especially in China. Methyltransferase-like 7 A (METTL7A), a member of the methyltransferase-like family, is associated with various cancers including CRC. Notably, CRC progression is closely linked to metabolic reprogramming. However, its precise role in CRC, particularly metabolic reprogramming of CRC, remains unclear. METTL7A was identified as a pivotal gene closely associated with CRC by bioinformatics analyses. Through a series of cellular functional assays and several animal experiments, such as in situ tumor and spontaneous tumor in C57BL/6 mice, the role of METTL7A in the development of colorectal cancer was evaluated. Transcriptomics and proteomics were used to analyze the effects of METTL7A on the expression of numerous genes, especially those involved in metabolic processes including cholesterol synthesis pathway within CRC cells. Western-blotting, co-immunoprecipitation and immunofluorescence were used to elucidate the relationship of METTL7A and the cholesterol metabolic pathway. METTL7A exhibited low expression in CRC cell lines and CRC tissues and it was demonstrated to function as a tumor suppressor in CRC. Transcriptomic and proteomic analyses indicated that METTL7A affects genes related to the cholesterol metabolism pathway. METTL7A was further proven to directly bind to Sterol Regulatory Element-Binding Protein1 (SREBP1) and SREBP Cleavage-Activating Protein (SCAP), hindering the nuclear translocation of SREBP1 and thereby reducing intracellular cholesterol content. This study provides valuable insights into the role of METTL7A in CRC and its impact on metabolic reprogramming, particularly cholesterol synthesis, and identifies METTL7A as a potential therapeutic target of CRC.

Saikogenin F (SGF) is a metabolite of Saikosaponin A (SSA) in vivo. However, in comparison to SSA, the neuroprotective efficacy and mechanisms of SGF remain uncertain in depression. The objective of this study was to explore the neuroprotective effects and mechanisms of SGF in corticosterone (CORT)-induced PC12 cells. Initially, analyses using MTT assays and flow cytometry demonstrated that SGF enhanced cell viability, inhibited cell death, and reduced levels of reactive oxygen species, lactate dehydrogenase and mitochondrial membrane potential. Furthermore, metabolomic analysis revealed that metabolic disorders were occurring in CORT-induced PC12 cells. SGF significantly reversed alterations in 13 metabolites and influenced 5 metabolic pathways. Of the five metabolic pathways, the regulation of purine metabolism is the most significantly affected by SGF. This study subsequently examined the regulatory impact of SGF on the P2X7R-NLRP3 and cAMP-PKA signaling pathways associated with purine metabolism, aiming to elucidate its neuroprotective mechanism. Enzyme-linked immunoassays and western blot analyses indicated that SGF significantly modulated the expression of proteins involved in these two pathways. These results show for the first time that SGF protected PC12 cells from damage caused by CORT through the regulation of the P2X7R-NLRP3 and cAMP-PKA signaling pathways in this study.

BACKGROUND:Anxiety is a common non-motor symptom of Parkinson's disease (PD), affecting 20 %-50 % of the patients, with an unmet need for effective therapies. Although perineuronal net (PNN) abnormalities have been implicated in both PD and anxiety, the underlying mechanisms and potential treatments remain poorly characterised. PURPOSE:In this study, we investigated whether baicalein ameliorates PD-associated anxiety via modulation of parvalbumin (PV) interneuron function by targeting semaphorin 3A (Sema3A). METHODS:Unilateral 6-hydroxydopamine (6-OHDA) lesioning of the substantia nigra pars compacta (SNc) was used to establish a PD rat model. Motor behaviours were assessed via gait analysis, rotarod test, and open field test (OFT). Anxiety-like phenotypes associated with PD were evaluated using the elevated plus maze (EPM) and marble burying test (MBT). The effects of baicalein on PNN level, PV neuron activity, and inhibitory synaptic markers (GABRA1, VGAT, and gephyrin) were examined via immunofluorescence (IF). Baicalein-Sema3A interactions were characterised through molecular docking, molecular dynamics simulation (MDS) and microscale thermophoresis (MST). The effects of baicalein on Sema3A/plexinA1 (PLXNA1)/neuropilin-1 (Nrp1) pathway were assessed using western blotting and qRT-PCR. Sema3A was knocked down to establish its link to PD-associated anxiety and PV interneuron dysfunction. RESULTS:Baicalein significantly ameliorated both anxiety-like behaviors and motor deficits in PD rats. It markedly reduced aggrecan expression and enhanced the inhibitory function of PV interneuron, as evidenced by up-regulation of GABRA1, VGAT, and gephyrin in the perirhinal cortex (PRh). Baicalein bound to Sema3A with high affinity (Kd = 0.86 ± 0.12 μM). Moreover, baicalein suppressed Sema3A/PLXNA1/Nrp1 signalling at both transcriptional and protein levels in the PRh. Furthermore, Sema3A knockdown in the PRh recapitulated the anxiolytic effects of baicalein by down-regulating aggrecan expression and rescuing the inhibitory function of PV interneuron, although its effects on motor deficits were limited. CONCLUSION:This study unravels a novel mechanism by which baicalein alleviates PD-associated anxiety probably via modulation of Sema3A-dependent PV interneuron dysfunction. The findings highlight the potential of Sema3A as a promising therapeutic target for PD-associated anxiety.