Neurochemical Research最新文献

The disruption of axonal continuity and neuronal loss at the injury site leads to structural disconnections as well as functional consequences in spinal cord injury. Herkinorin, a structurally unique non-nitrogenous opioid receptor agonist, demonstrates significant neuroprotective efficacy in models of ischemic brain injury and epilepsy. In this report, we aimed to explore the role and underlying mechanism of Herkinorin in axon regeneration and functional recovery in SCI. Oxygen–glucose deprivation and restoration (OGD/R) condition was used in cell line rat pheochromocytoma cells (PC12) and primary cortical neurons to mimic SCI context in vitro. and apoptosis of the cells were detected. A rat SCI model was conducted by Allen’s impactor. Cells were treated with Herkinorin under OGD/R condition. Footprint analysis and BBB scale was to evaluate locomotor function. Western blot and immunofluorescence staining of proteins Map2, GAP43 and Ace-tubulin to assess microtubule stabilization and Bax, Bad, Bcl2 as well as cleaved caspase3 to detect apoptosis. The TNFα was employed to determine the expression profiles of NF-κB. Herkinorin alleviated OGD/R-induced neuronal apoptosis and axonal damage in PC12 cells and primary neurons by enhancing microtubule stability and inhibiting NF-κB, effects abolished by TNFα. Crucially, in a rat SCI model, Herkinorin improved locomotor function (BBB scores, gait) and mitigated spinal cord neuronal loss and microtubule disassembly. Herkinorin confers neuroprotection and pro-regenerative effects against neuronal injury both in vitro and in vivo, mechanistically linked to microtubule stabilization and NF-κB pathway suppression, positioning it as a promising candidate for SCI therapy.

Age-related cognitive decline is a major public health concern, with few effective interventions available. Intermittent fasting (IF) has emerged as a promising metabolic intervention, potentially enhancing neuroprotection through increased β-hydroxybutyrate (BHB) production. This study aimed to test the hypothesis that IF attenuates cognitive decline in D-galactose (D-gal)-induced aging rats, at least partly in association with BHB elevation and modulation of PI3K/AKT/GSK-3β signaling. Forty-six male Sprague-Dawley rats (2 months old) were assigned to five groups: Control, D-gal-induced aging (D-gal), D-gal + IF, D-gal + IF + LY294002 (a PI3K inhibitor), and D-gal + IF + Solvent. IF was administered for 6 weeks in conjunction with D-gal treatment. Recognition memory and spatial learning/memory were assessed using the novel object recognition and Morris water maze tests, respectively; the open field test was used to evaluate locomotor activity and anxiety-like behavior. Biochemical assays (ELISA, Western blot), histological staining, and RNA-seq transcriptomic analysis were performed. IF improved recognition memory and spatial learning/memory, preserved hippocampal neuronal morphology, increased the proportion of morphologically normal CA1 neurons, elevated serum BHB levels, reduced neuroinflammation and tau phosphorylation, and restored brain-derived neurotrophic factor (BDNF) expression. These improvements were attenuated by LY294002 intervention, supporting the involvement of the PI3K/AKT/GSK-3β pathway. Transcriptomic analysis revealed suppression of senescence-related genes and modulation of neuroprotective pathways. IF attenuated cognitive decline in D-gal-induced aging rats in association with increased BHB levels and modulation of the PI3K/AKT/GSK-3β pathway. These findings suggest that IF may represent a promising non-pharmacological intervention for age-related cognitive dysfunction, although causal roles of ketone metabolism require further validation.

This study aimed to investigate the regulatory role of High-mobility group box 1 (HMGB1) in neuronal ferroptosis following spinal cord injury (SCI) and its underlying mechanisms. Iron ion deposition, malondialdehyde (MDA) and glutathione (GSH) levels, as well as the expressions of HMGB1, acyl-CoA synthetase long-chain family member 4 (ACSL4), solute carrier family 7 member 11 (SLC7A11), and glutathione peroxidase 4 (GPX4) in spinal cord tissue of SCI rats were measured at 24 h, 72 h and 1 week post-injury. Then, an in vitro neuronal ferroptosis model was established and neuronal cells were transfected with lentiviral vectors for HMGB1 interference or ACSL4 overexpression. Iron ion levels, MDA content, GSH activity, and the expressions of HMGB1, ACSL4, SLC7A11 and GPX4 were measured. And the interaction between HMGB1 and ACSL4 was assessed. Finally, SCI rats were administered the HMGB1 inhibitor glycyrrhizic acid (GA) and the effects GA on the iron ion deposition, MDA and SOD levels, as well as the expressions of HMGB1, ACSL4, SLC7A11 and GPX4 in spinal cord tissues were evaluated. SCI induced time-dependent iron deposition, increased MDA, HMGB1, and ACSL4, and decreased GSH, GPX4, and SLC7A11. The administration of GA in SCI rats significantly reduces iron ion deposition, decreases the levels of MDA, HMGB1 and ACSL4, and increases the levels of GSH, GPX4, and SLC7A11. Furthermore, cellular-level results demonstrated that interfering with HMGB1 attenuated ferroptosis in rat spinal cord neurons by suppressing ACSL4. Targeted suppression of ACSL4 expression through interference with HMGB1 inhibits neuronal ferroptosis in SCI rats.

Ethylmalonic (EMA) and methylsuccinic (MSA) acids concentrations are elevated in tissues and body fluids of patients with ethylmalonic encephalopathy (EE), a mitochondrial disorder associated with basal ganglia abnormalities. To clarify the pathophysiology of this disorder, we evaluated the effects of EMA and MSA on bioenergetics, redox homeostasis and mitochondrial permeability transition (MPT) in rat striatum. We verified that EMA and MSA reduced state 3 and uncoupled respiration and inhibited the activities of α-ketoglutarate dehydrogenase and complex IV of the electron transport chain. Both organic acids also decreased mitochondrial membrane potential and Ca²⁺ retention capacity, which were normalized by cyclosporine A and ADP, indicating induction of MPT pore opening. These effects were further mitigated by N-ethylmaleimide and dithiothreitol, suggesting that thiol groups of the MPT pore were oxidized by EMA and MSA. Moreover, EMA and MSA mildly decreased reduced glutathione concentrations, reinforcing that thiol group are oxidized by these organic acids. Therefore, it is presumed that EMA- and MSA-induced bioenergetic impairment associated with MPT pore opening is involved in the pathophysiology of basal ganglia injury observed in EE.

Glioblastoma (GBM) is the most aggressive primary brain tumor with a dismal prognosis. Ferroptosis is implicated in GBM pathogenesis. Heat shock protein B1 (HSPB1) is associated with tumor progression, yet its precise function and regulatory mechanism in GBM ferroptosis remain elusive. Differentially expressed genes were identified from the GSE151352 dataset. WGCNA was employed to identify GBM-associated modules, which were then intersected with genes from the FerrDb V2 database. HSPB1 expression and prognostic value were validated using TCGA and GEPIA databases, and clinical specimens. Functional assays (EdU, TUNEL, and Transwell) and ferroptosis indicators (lipid ROS, Fe²⁺, GSH) were assessed following HSPB1 modulation. Bioinformatics tools predicted METTL1-mediated m7G modification of HSPB1, and results were validated by RIP, dual-luciferase reporter assay, and mRNA stability assays. Transcriptional regulation of HSPB1 by HOXA5 was predicted and confirmed. A subcutaneous xenograft model was used to evaluate the METTL1-HSPB1 axis in vivo. Analysis revealed 2985 DEGs. WGCNA identified a GBM-correlated “red” module; intersection with ferroptosis genes pinpointed HSPB1. HSPB1 was significantly overexpressed in GBM, correlating with poor patient survival. HSPB1 knockdown suppressed GBM cell proliferation, migration, invasion, and induced ferroptosis. Mechanistically, METTL1 mediated m7G modification to HSPB1 mRNA to enhance its stability. Concurrently, HOXA5 bound to the HSPB1 promoter to activate its transcription. Silencing either METTL1 or HOXA5 downregulated HSPB1, inhibiting GBM malignant phenotypes. In vivo, the METTL1-HSPB1 axis promoted tumor growth. METTL1 stabilizes HSPB1 mRNA through m7G methylation, and HOXA5 transcriptionally activates HSPB1 expression. This regulation promotes GBM malignant progression.

Graphical Abstract

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Bioinformatic screening identifies HSPB1 as a ferroptosis-related hub gene in GBM: METTL1 stabilizes HSPB1 mRNA via m7G modification, while HOXA5 transcriptionally activates HSPB1 by binding its promoter. Elevated HSPB1 drives GBM proliferation, invasion, and resistance to apoptosis and ferroptosis, promoting tumor malignancy.

Forty years ago, in 1986, Rita Levi-Montalcini was awarded the Nobel Prize in Physiology or Medicine for the discovery of Nerve Growth Factor, a fundamental finding that inaugurated the field of trophic signaling in the nervous system. Her research, conducted with credibility and consistency under the precarious conditions of an Italy enduring the Second World War, and later refined in the United States, rigorously demonstrated that neuronal growth and survival are regulated by specific chemical signals, not merely by intrinsic processes. Using biological assays involving sarcoma and chicken embryos, Levi-Montalcini identified a protein substance that promoted exuberant nerve fiber growth from sensory and sympathetic ganglia. The subsequent purification and biochemical characterization of Nerve Growth Factor by biochemist Stanley Cohen — with whom she shared the Nobel Prize — confirmed its protein nature. This discovery scientifically proved the existence of signaling molecules that guide nervous system development, including critical processes such as neuronal differentiation, axonal growth, and apoptosis, programmed cell death. The concept that target-cells secrete trophic factors for maintenance and survival became fundamental in neuroembryology. The impact of Levi-Montalcini’s work transcended developmental neurobiology. The Nerve Growth Factor model paved the way for the discovery of a family of neurotrophins, elucidating mechanisms of neural plasticity, regeneration, and brain aging, with profound implications for the understanding of mental illnesses, neurodegenerative diseases, and cancer. For establishing, with experimental precision, a universal principle of cellular communication in the nervous system, Rita Levi-Montalcini — a scientist who expanded scientific frontiers — was honored with numerous awards, alongside the Nobel Prize, consolidating her position as an influential female figure in 20th-century neuroscience.

Dysregulation of the WNT signaling pathway is implicated in the synaptic dysfunction underlying Alzheimer’s disease (AD). This study investigates whether enhancing WNT signaling through hippocampal overexpression of the WNT3a ligand can mitigate functional deficits in two distinct animal models of AD pathology. We used a rat model of cholinergic deficit, induced by intraseptal 192IgG-saporin injections, and a transgenic 5XFAD mouse model of amyloidosis. Adeno-associated viruses were used to overexpress WNT3a in the hippocampal CA1 region. We assessed cognitive and sensorimotor behavior, synaptic plasticity (long-term potentiation, LTP) in vivo (rats) and in vitro (mice), and analyzed key proteins of the WNT signaling pathway. In rats with cholinergic deficit, WNT3a overexpression ameliorated sensorimotor coordination deficits and restored the initial phase of hippocampal LTP in vivo, without preventing the loss of cholinergic neurons or the decrease in acetylcholinesterase activity. In 5XFAD mice, which exhibited impaired LTP in vitro, WNT3a overexpression significantly enhanced the early phase of potentiation. This functional rescue was associated with a recovery of elevated phospho-β-catenin levels in the 5XFAD hippocampus. WNT3a did not affect behavior in the 5XFAD model and its benefits were independent of changes in cholinergic markers in both models. Our findings demonstrate that targeted WNT3a overexpression in the hippocampus can partially alleviate synaptic and functional deficits in AD models by directly modulating synaptic plasticity, primarily through the restoration of the Wnt/β-catenin pathway. This positions WNT3a gene therapy as a promising strategy for counteracting synaptic failure in Alzheimer’s disease.

Ischemic stroke poses a severe threat to human health, and cerebral ischemia-reperfusion injury (CIRI) following recanalization remains a critical challenge for patient prognosis. This study investigated the role and mechanism of the circ_0017866/miR-124-3p/claudin domain containing 1 (CLDND1) axis in CIRI-induced endothelial-mesenchymal transition (EndMT) and evaluated the therapeutic potential of lipid nanoparticle (LNP)-encapsulated circ_0017866. Both oxygen-glucose deprivation/reperfusion cell and middle cerebral artery occlusion/reperfusion mouse models were established. Molecular interactions and gene/protein expression were validated using dual-luciferase reporter assays, quanti-tative real-time PCR, western blot, and immunofluorescence. In vitro, endothelial cell viability and function were assessed through CCK-8, Transwell migration, and tube formation assays. In vivo, LNP-mediated delivery of circ_0017866 was employed, and neurological deficits and cerebral infarct volume were evaluated using modified neu-rological severity scores and 2,3,5-triphenyltetrazolium chloride staining. Both in vivo and in vitro studies demonstrated that after ischemia-reperfusion, the expression of circ_0017866 and CLDND1 was downregulated, while miR-124-3p expression was upregulated. The dual-luciferase assay confirmed that miR-124-3p directly targets both circ_0017866 and CLDND1. In the OGD/R model, either circ_0017866 overexpression or miR-124-3p inhibition upregulated the expression of CLDND1 and occludin, downregulated α-smooth muscle actin (α-SMA) and Snail, and improved endothelial function. In the MCAO/R model, LNP-circ_0017866 treatment significantly improved neurological deficits, reduced infarct volume, and decreased the abnormal expression of EndMT-related markers. Circ_0017866 alleviates CIRI-induced EndMT by functioning as a molecular sponge for miR-124-3p to indirectly regulate CLDND1 expression. LNP-delivered circ_0017866 effectively mitigates neurological deficits and reduces cerebral infarct volume, providing a promising therapeutic strategy for ischemic stroke.

Alzheimer’s disease (AD) is characterized by neurodegeneration, autophagy dysregulation, and mitochondrial stress. β-asarone and tenuifolin have shown neuroprotective effects, but their mechanisms remain unclear. YuanZhi decoction, a traditional formula containing Polygala tenuifolia, is used for cognitive impairment, yet its active constituents are not fully understood. A D-galactose-induced AD mouse model (150 mg/kg/day, s.c., 42 days) was established to evaluate β-asarone, tenuifolin, their combination, and YuanZhi decoction. Behavioral tests (MWM and NOR), Nissl staining, IHC, network pharmacology, molecular docking (including the GRIN2B inhibitor ifenprodil as positive control), Western blotting, biochemical assays, and RT-qPCR were performed. All treatments improved cognitive function, attenuated hippocampal neuronal loss and tau pathology, and restored metabolic parameters (ATP, ROS, DT, SOD2). Network pharmacology identified GRIN2B as a key hub target. Molecular docking revealed that β-asarone and senegenin (the active metabolite of tenuifolin) bind to the same allosteric pocket as ifenprodil on GRIN2B, sharing highly consistent interaction residues. GRIN2B upregulation in AD mice was accompanied by autophagic dysfunction (increased LC3B/LC3A ratio and PINK1, reduced p62) and mitochondrial stress. These abnormalities were significantly reversed by all treatments, with the combination and YuanZhi decoction showing greater efficacy. β-asarone, tenuifolin, their combination, and YuanZhi decoction alleviate GRIN2B-linked autophagic imbalance and mitochondrial stress in AD-like mice, supporting their therapeutic potential.

Graphical Abstract

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Individuals with hypertension carry a high risk of stroke, which endangers human health. The study systematically elucidated the dynamic expression, functional effects, and molecular mechanisms of the circular RNA LPAR3 in ischemic stroke (IS). 300 essential-hypertension (EH) patients were enrolled, comprising 165 cases diagnosed with IS. mRNA abundance was quantified via RT-qPCR. C57BL/6 mice treated with MACO and Neuro-2a cells receiving oxygen-glucose deprivation/reoxygenation (OGD/R) were applied for functional experiments. Neurological deficits and infarct volume of mice were evaluated. Besides, key cellular behaviors encompassing proliferative capacity, apoptotic rate, and inflammatory response were monitored. The putative binding event was interrogated through luciferase reporter and RIP coupled to qPCR assays. A diminished level of circLPAR3 was evidenced in the plasma of IS patients, which can distinguish IS cases from EH and remains an independent predictor of patients’ poor prognosis. In vivo, overexpression of circLPAR3 alleviated MACO-mediated neurological deficit and cerebral infarction. In vitro, OGD/R triggered the surge in cell apoptosis and pro-inflammatory cytokine release, but it was attenuated by circLPAR3 upregulation. Mechanistically, the neuro-rescue driven by circLPAR3 was offset by miR-634, whose plasma expression was negatively correlated with circLPAR3. KLB expression was significantly downregulated in cellular models and co-regulated by circLPAR3 and miR-634. circLPAR3/miR-634 modulates the pathological progression of IS by directly regulating neuronal apoptosis and orchestrating inflammatory responses, and KLB may serve as a key downstream target mediating the biological functions.