Social isolation and loneliness can lead to an increased risk of cardiovascular disease, cognitive decline, and psychiatric disorders. In this study, we investigated the impact of chronic social isolation on the cognitive functions and structural-functional state of the brain in middle-aged rats. Ten-month-old male spontaneously hypertensive (SHR), Wistar-Kyoto (WKY), and Wistar rats were housed individually for 15 weeks, while the control animals of respective strains were housed in groups. After 13 weeks, all the rats were trained to complete a spatial task in a Barnes maze. After the behavioral experiments, we analyzed the density of neurons, astrocytes, and microglia in the hippocampus. We also studied the expression of several genes in the hippocampus and frontal cortex, including Aqp4, Cx3cr1, Tnfa, Il1b, and Il6, using qPCR. We found that Wistar rats showed high resilience, maintaining stable physiological parameters and spatial learning abilities without significant neuroinflammatory or structural changes in the brain. In contrast, SHRs showed marked physiological vulnerability, characterized by a significant weight loss and severe memory impairments in the Barnes maze, accompanied by an elevated cytokine response in both brain regions studied. A different trajectory of neurobiological changes was observed in WKY rats. Although they had a high rate of hippocampal neurogenesis, which provided a compensatory effect on basic spatial learning and memory, they also showed decreased neuronal density and an elevated Il6 mRNA level, linked to impaired microglial Cx3cr1 mRNA expression. This study highlights the importance of genetic background in determining the outcome of brain aging in conditions of social isolation.
Lipopolysaccharide (LPS)-induced endotoxemia is widely used to model neuroinflammation-associated cognitive impairment, primarily involving activation of Toll-like receptor 4 (TLR4) and downstream NF-κB signaling. This study investigated whether probiotic interventions with Saccharomyces boulardii or Lactobacillus rhamnosus attenuate LPS-induced neurobehavioral and molecular alterations. Male Wistar rats were exposed to subchronic intraperitoneal LPS (0.25 mg/kg/day, 9 days) with or without concurrent probiotic administration. Behavioral assessments included the open field, elevated plus maze, forced swim, and novel object recognition tests. Hippocampal levels of inflammatory (TLR4, NF-κB, TNF-α, IL-6, GFAP), neurotrophic (BDNF, CREB), cholinergic (ACh, AChE), and apoptotic (p53, BAX, caspase-3) markers were quantified by ELISA. LPS exposure was associated with increased hippocampal TLR4 and NF-κB p65 levels, elevated TNF-α and IL-6 levels, and higher GFAP levels, alongside reduced BDNF and CREB levels, an altered ACh/AChE balance, and increased levels of apoptosis-related proteins. These molecular alterations coincided with deficits in recognition memory and increased anxiety- and depressive-like behaviors without affecting locomotor activity. Both S. boulardii and L. rhamnosus significantly attenuated these changes, reducing inflammatory markers, improving neurotrophic and cholinergic parameters, and partially attenuating apoptosis-related proteins. Behavioral impairments were concurrently ameliorated. Overall, these findings suggest that LPS-induced cognitive and behavioral alterations are closely associated with coordinated inflammatory, neurochemical, and apoptotic changes in the hippocampus. Because both probiotics were administered prophylactically, these findings indicate preventive rather than therapeutic efficacy. As all molecular outcomes were quantified by ELISA in whole-hippocampus homogenates, they should be interpreted as changes in protein levels rather than as demonstrated pathway activation.
Ischemia-reperfusion (IR) injury is characterized by significant neuronal apoptosis and mitochondrial damage.Despite significant advancements in understanding the pathophysiological mechanisms of IR, effective therapeutic strategies remain limited. Previous studies have shown that mild hypothermia (MH) can significantly alleviate brain IR injury by reducing neuronal apoptosis and mitochondrial damage, while mitophagy plays a crucial role in maintaining cellular self-protection and mitochondrial homeostasis. However, the involvement of the PINK1/Parkin signaling pathway, a classical regulatory route for mitophagy, in the protective effects of MH against IR remains unclear. To investigate this phenomenon, we used neuron-like SH-SY5Y cells and employed an oxygen-glucose deprivation/reoxygenation (OGD/R) model to simulate the in vitro IR process, and conducted a series of cell experiments. Our results demonstrated that under MH conditions, the expression levels of PINK1 and Parkin were upregulated after 5 h of ischemia-hypoxia followed by 24 h of reoxygenation, and the protective effect on cells was also significant. Furthermore, the inhibition of the mitophagy pathway may partially eliminate the protective effect provided by MH against OGD/R-induced damage.These findings indicate that the protective effects of MH against IR is mediated, at least in part, through the PINK1/Parkin-dependent mitophagy pathway.This study provides new insights into the mechanisms by which MH mitigates IR injury and may offer potential therapeutic targets for treating IR-related diseases.
Sepsis-associated encephalopathy (SAE) is a devastating condition with high mortality and cognitive sequelae, yet no effective treatments. This study investigates the neuroprotective potential of selective Sirtuin 1 (SIRT1) activation using the synthetic agonist SRT1720 in a male C57BL/6 J murine model of SAE. Transcriptomic profiling of the hippocampus revealed a specific disruption in glutathione and arachidonic acid metabolism pathways, indicating a metabolic priming for ferroptosis. We demonstrate that SIRT1 activation improves survival and ameliorates cognitive deficits by concurrently attenuating hippocampal ferroptosis and neuroinflammation. We observed that this protection was associated with the upregulated protein expression of Nrf2 and GPX4, key regulators of ferroptosis. Mechanistically, SIRT1 activation suppressed hippocampal ferroptosis, marked by reduced lipid peroxidation (ROS and MDA). Concurrently, it attenuated neuroinflammation by suppressing microglial activation and NLRP3 inflammasome activity, thereby decreasing the maturation of proinflammatory cytokines. These findings suggest SIRT1 activation coordinately counteracts both ferroptotic and inflammatory damage in SAE, presenting a compelling therapeutic strategy.
Neuroinflammation is a response of the central nervous system to injury, infection, or disease, involving the activation of glial cells and the release of cytokines and reactive species. When sustained or dysregulated, it contributes to the progression of neurological disorders. Quercetin (Que) is one of the most extensively studied flavonoids; however, its therapeutic application is limited by low bioavailability. Glucosylation has therefore emerged as a strategy to improve the physicochemical and biological properties of flavonoids. Previously, we synthesized the derivative quercetin-3’-α-D-glucoside (Que-3α) and characterized its pharmacokinetic profile and in vitro cytotoxicity. Here, we investigated the anti-inflammatory and antioxidant effects of Que and its derivatives, including the recently synthesized Que-3α and the naturally occurring β-glucoside, quercetin-3-β-D-glucoside (Que-3β), in a lipopolysaccharide (LPS)-induced inflammation model using C6 astroglial cells. Cells were pretreated for 2 h with the compounds (15 and 30 µM) and then stimulated with LPS (10 µg/mL) for 24 h. The compounds preserved mitochondrial activity, while Que and Que-3α reduced LPS-induced reactive oxygen species production, increased cell viability, and decreased apoptosis and necrosis. Notably, the Que-3α reduced iNOS expression, indicating more effective modulation of inflammatory mediators compared with the aglycone. These findings reinforce the biological properties of Que and suggest that α-glucosylation preserves the beneficial actions of Que while enhancing pharmacodynamic features relevant to neuroinflammation.
The growing prevalence of obesity has been associated with metabolic and neurobiological alterations that compromise brain integrity and behavior. Diet-induced obesity, often accompanied by hypercholesterolemia, activates apoptotic signaling pathways in the central nervous system, contributing to cognitive and behavioral impairments. This study evaluated the effects of atorvastatin on behavioral performance and apoptotic signaling pathways in the hippocampus and cerebral cortex of rats subjected to a hypercaloric cafeteria diet. Male Wistar rats were assigned to four groups: control diet (CTR), control diet plus atorvastatin (ATOR), cafeteria diet (CAF), and cafeteria diet plus atorvastatin (CAF+ATOR), and exposed to the diets for 24 days. Body weight, adiposity, and circulating cholesterol levels were measured to validate the metabolic model. Behavioral assessments included locomotor activity, immobility time, and recognition memory. The expression of pro-caspase−8, pro-caspase−9, and apoptosis-inducing factor (AIF) was quantified in frontal cortex and hippocampus. The cafeteria diet increased adiposity and circulating cholesterol levels, confirming metabolic dysfunction, whereas atorvastatin reduced cholesterol levels and prevented retroperitoneal fat accumulation. In addition, the cafeteria diet altered the expression of apoptosis-related proteins in the hippocampus and cortex, whereas atorvastatin attenuated these changes. Cafeteria-fed rats showed reduced exploratory activity and increased immobility. Spatial memory was impaired by both atorvastatin and the cafeteria diet but preserved in the CAF+ATOR group. Novel object recognition was impaired only in the ATOR group. These findings indicate that atorvastatin exerts context-dependent effects under conditions of diet-induced metabolic dysfunction.
Spontaneous cerebral hemorrhage (sICH) is one of the cerebrovascular diseases with the highest mortality rate. MicroRNAs (miRNAs) are implicated in the regulation of various pathological processes, however, the specific function of miR-642b-3p in sICH remains unclear. Serum miR-642b-3p levels of sICH patients was determined. A thrombin-induced human brain microvascular endothelial cell (hBMECs) injury model was established. Cell viability, apoptosis, inflammatory factors, and related protein expression were assessed using CCK-8 assay, flow cytometry, ELISA, and Western blot. The targeting relationship between miR-642b-3p and CCNA2 was validated via a dual-luciferase reporter assay. MiR-642b-3p in the serum of ICH patients was significantly increased. Following thrombin induction, miR-642b-3p was upregulated in hBMECs, accompanied by decreased cell viability, increased apoptosis, elevated production of inflammatory factors, upregulated expression of pro-apoptotic proteins (Bax and cleaved caspase-3), and downregulated expression of anti-apoptotic protein (Bcl-2). Additionally, the protein levels of tight junction markers ZO-1 and occludin were markedly decreased. Notably, inhibition of miR-642b-3p significantly reversed these thrombin-induced changes and alleviated cell damage. Mechanistically, CCNA2was confirmed as a direct downstream target gene of miR-642b-3p. MiR-642b-3p is upregulated in sICH and mediates thrombin-induced endothelial cell injury, at least in part, by targeting CCNA2. These findings suggest that the miR-642b-3p/CCNA2 axis may represent a potential mechanistic pathway involved in ICH-induced blood-brain barrier disruption.
Epilepsy, a common neurological disorder, is distinguished by abnormal neuronal activity. This study explored the mechanism of the long non-coding RNA (lncRNA) GABPB1-AS1/SLC12A5 pathway modulating neuronal ferroptosis during epileptic seizures. Loss- and gain-of-function approaches were conducted in a mouse model of kainic acid-induced seizures and glutamic acid (Glu)-treated HT22 neurons to explore the regulatory effect of lncRNA GABPB1-AS1/SLC12A5 on neuronal ferroptosis. Mouse seizures were assessed using the Racine scale. LncRNA GABPB1-AS1, SLC12A5, and ferroptosis-related markers were measured via RNA quantification, Western blot, and immunofluorescence. The direct or indirect interaction between lncRNA GABPB1-AS1 and SLC12A5 was validated using RIP, RNA pull-down, and Co-IP assays. CHX chase and ubiquitination assays assessed SLC12A5 stability and ubiquitination levels regulated by lncRNA GABPB1-AS1. LncRNA GABPB1-AS1 was upregulated in the hippocampal CA1 region of KA-induced mice. LncRNA GABPB1-AS1 knockdown suppressed neuronal ferroptosis-lowering Fe2+, MDA, and lipid peroxidation, while raising GSH, SLC7A11, and GPX4. Its knockdown also reduced seizure scores and frequency, prolonged seizure latency, and shortened seizure duration. An in vitro experiment verified the suppressive effect of lncRNA GABPB1-AS1 knockdown on neuronal ferroptosis. In addition to a direct binding relationship, this lncRNA further recruited the E3 ligase PRKN to enhance SLC12A5 ubiquitination and proteasomal degradation. SLC12A5 overexpression inhibited Glu-induced ferroptosis, and SLC12A5 knockdown partially reversed the effects exerted by lncRNA GABPB1-AS1. LncRNA GABPB1-AS1 recruits the E3 ligase PRKN to promote ubiquitination and proteasomal degradation of SLC12A5, thereby downregulating SLC12A5 expression and facilitating hippocampal neuronal ferroptosis that mediates epileptic seizures.
Age-related insomnia (ARI) is becoming increasingly prevalent among the elderly and remains inadequately addressed by conventional pharmacotherapy, prompting growing interest in stem cell–based alternatives. The present study investigated whether exosomes derived from umbilical cord mesenchymal stem cells (UCMSC-Exos) ameliorate ARI by suppressing hippocampal neuronal ferroptosis. UCMSCs were isolated from Sprague–Dawley rats on gestational day 21, and UCMSC-Exos were purified from passage 3 supernatants by ultracentrifugation. Transmission electron microscopy, nanoparticle tracking analysis, and western blotting confirmed characteristic exosomal morphology with a mean diameter of 118.4 nm, positive expression of TSG101, CD9, and CD81, and absence of Calnexin. Following tail vein injection of DiD-labeled UCMSC-Exos in rats, DiD signals were traceable for 21 days. An ARI model was established in adult male rats using D-galactose combined with para-chlorophenylalanine, and animals were randomized into Sham, ARI, ARI-UCMSC-Exos, and ARI-Fer-1 groups. Morris water maze, elevated plus maze, open field tests, and electroencephalography demonstrated that UCMSC-Exos reversed behavioral deficits and prolonged total and slow-wave sleep. Hematoxylin-eosin, Nissl, and Golgi staining revealed restored dentate gyrus neurons with increased dendritic number and length, while transmission electron microscopy showed improved synaptic and mitochondrial ultrastructure. Prussian blue staining, ELISA, and western blotting further indicated that UCMSC-Exos reduced Fe3+, glutamate, and reactive oxygen species while elevating cysteine, glutathione, GPX4, 5-HT, and BDNF. Collectively, UCMSC-Exos alleviate sleep disturbance in ARI rats by inhibiting hippocampal neuronal ferroptosis, highlighting their therapeutic potential for ARI.
Postoperative neurocognitive disorders (PND) are common complications in older surgical patients, but druggable mechanisms linking surgical trauma to hippocampal synaptic and cognitive vulnerability remain poorly defined. The σ-2 receptor (σ-2R), encoded by transmembrane protein 97 (TMEM97), and progesterone receptor membrane component 1 (PGRMC1) intersect membrane lipid biology and synaptic stress signaling; CT1812, a pharmacological σ-2R complex modulator, has shown target engagement in early Alzheimer’s disease trials, but whether this pathway participates in postoperative hippocampal injury is unknown. This study examined whether surgery-induced hippocampal stress in aged mice is associated with TMEM97 and PGRMC1 signaling and whether early CT1812 treatment can attenuate this response. Young adult (2–3 months) and aged (18–20 months) male C57BL/6J mice underwent sham operation or laparotomy; aged mice were further treated with vehicle or CT1812 (3 mg/kg, intravenous injection, days 0–3). Hippocampal molecular changes were assessed by western blotting and immunofluorescence on day 3, and behavior was evaluated by open field test (OFT) and Morris water maze (MWM) on days 6–10. Laparotomy increased hippocampal TMEM97 and PGRMC1 expression and induced anxiety-like and spatial-memory deficits preferentially in aged mice, without evidence that locomotor impairment explained the behavioral phenotype. TMEM97 and PGRMC1 were detected in neuronal nuclei (NeuN)-positive neurons, ionized calcium-binding adapter molecule 1 (Iba-1)-positive microglia, and glial fibrillary acidic protein (GFAP)-positive astrocytes in hippocampal cornu ammonis 1 (CA1), supporting multicellular pathway involvement. In aged surgical mice, CT1812 significantly reduced upregulation of TMEM97 and PGRMC1, improved OFT center-zone occupancy and MWM performance, and restored the synaptic markers postsynaptic density protein 95 (PSD95) and synaptophysin. CT1812 also normalized lipid-handling proteins, including apolipoprotein E (ApoE), ATP-binding cassette transporter A1 (ABCA1), cholesterol 24-hydroxylase (CYP46A1), and low-density lipoprotein receptor (LDLR), and reduced perilipin 2 (PLIN2) accumulation in neurons and microglia. In parallel, CT1812 attenuated a selective endoplasmic reticulum (ER) stress response involving binding immunoglobulin protein (BIP) and protein kinase R-like endoplasmic reticulum kinase (PERK)–eukaryotic initiation factor 2α (eIF2α) signaling in hippocampal neurons, and suppressed microglial nuclear factor κB (NF-κB)/NOD-like receptor family pyrin domain containing 3 (NLRP3) inflammatory activation, whereas C/EBP homologous protein (CHOP) was not significantly changed. These findings support a model in which postoperative injury in the aged hippocampus engages a lipid-proteostatic and inflammatory stress program associated with TMEM97 and PGRMC1, and suggest that CT1812 attenuates cognitive vulnerability by restraining this coupled response.
Collagen is a major extracellular matrix component, and soy protein has been reported to influence cellular and immune-related processes. Nanofiber scaffolds incorporating collagen and soy protein isolate (SPI) may provide a platform for modulating cell–material interactions in neural systems. In this study, we fabricated electrospun nanofibers composed of collagen (CO), SPI, and polycaprolactone (PCL) and investigated the cellular and transcriptional responses of human astrocytes to these scaffolds in vitro. The nanofibers were characterized by scanning electron microscopy (SEM), Fourier transform infrared (FTIR) spectroscopy, and contact angle analysis. Human fetal astrocytes exhibited high viability on all nanofiber scaffolds. Flow cytometry analysis indicated that incorporation of SPI into CO/PCL nanofibers did not alter cell cycle distribution. Aligned nanofibers provided directional guidance for astrocyte migration. RNA-sequencing analysis revealed enrichment of the “neurodegeneration” and “antigen processing and presentation” pathways among the down-regulated genes in cells on CO/SPI/PCL fibers compared with CO/PCL fibers. Down-regulated genes in these pathways include IL1B, IL6, HLA-B, HLA-DMB, HLA-DPA1, and HLA-DRA. The “focal adhesion” pathway is enriched among up-regulated genes, which include COL4A1, COL4A2, FN1, LAMB1, LAMB2, AKT2, RAC1, RAC2, ROCK2, and PIP5K1A. These results demonstrate that incorporation of SPI into collagen-based nanofibers modulates astrocyte migration and gene expression profiles associated with focal adhesion and immune-related pathways, providing a foundation for further investigation of SPI-containing biomaterials in neural tissue engineering applications.
Polytherapy is increasingly needed to overcome the limitations of monotherapy in epilepsy and its comorbidities. Therefore, we investigated the effect of cenobamate (CNB, 30 mg/kg) and trazodone (TZ, 10 mg/kg), alone and in combination, in PTZ-kindling model, where mice received 11 doses of pentylenetetrazole (PTZ, 40 mg/kg) every other day for three weeks. Behavioral seizure scoring, electroencephalographic (EEG) changes at the 1st, 5th, and 11th injections, molecular markers, and hippocampal histopathology were assessed. TZ + CNB suppressed seizure progression with no tonic-clonic seizures observed throughout kindling period (p < 0.0001), whereas monotherapy by these drugs showed no significant suppression. PTZ produced the most pronounced EEG abnormalities, which CNB and TZ alone did not differ significantly. Combination therapy revealed the greatest reductions across all EEG parameters (q = 0.0004–0.0014), approaching near-baseline values by the 11th injection. Furthermore, brain homogenates were assessed for BDNF, GFAP, and TrkB expression levels by RT-PCR. CNB showed mixed effects (p = 0.0081–0.0631), while TZ (p = 0.0971–0.1422) produced no significant difference from PTZ40. TZ + CNB shifted these markers toward normal levels (p = 0.0001–0.0054), outperforming monotherapies. Hippocampal Nissl-stained sections showed neurodegenerative damage with monotherapies across all regions, while TZ + CNB exhibited substantial preservation, with minimal alteration in hippocampal and cortical architecture. These parallel findings suggest that positive allosteric GABAA modulation and persistent sodium-current inhibition by CNB, combined with TZ serotonergic modulation, reduce hyperexcitability, pathological neuroplasticity, and glial reactivity. Collectively, TZ + CNB is a promising mechanistically complementary regimen that suppresses seizure progression and associated molecular and neurostructural pathology, which needs further preclinical optimization and translational investigations.
Parkinson’s disease (PD) is a neurodegenerative disease characterized by progressive loss of dopaminergic neurons. Studies have shown that mesenchymal stem cells (MSCs) have prominent neuroprotective potential to improve PD. Previous study found that dental pulp‑derived MSCs, including stem cells from human exfoliated deciduous teeth (SHED), significantly ameliorated PD-related pathology, but the underlying molecular mechanism remains unclear. In this study, HSPB1 and PARK7 were initially identified through transcriptomic analyses of mouse and clinical samples, immunoprecipitation‑mass spectrometry (IP‑pulldown), and protein‑protein interaction (PPI) network analysis. Using an MPTP-induced SH-SY5Y cell model of PD, we found that SHED treatment alleviated oxidative stress, restored mitochondrial membrane potential, and inhibited apoptosis, accompanied by significant upregulation of HSPB1 and PARK7. Using siRNA interference, we confirmed that both HSPB1 and PARK7 are involved in the improvement of PD by SHED. Rescue experiments showed that HSPB1 overexpression partially restored the neuroprotective effects impaired by PARK7 knockdown, whereas PARK7 overexpression failed to compensate for HSPB1 deficiency, suggesting a functional dependence between PARK7 and HSPB1 in SHED-mediated neuroprotection. This study demonstrates that SHED improves PD pathological progression through the PARK7/HSPB1-related mechanism and indicates that HSPB1 may serve as a potential therapeutic target for PD. SHED-mediated neuroprotection in PD models is associated with upregulation of PARK7 and HSPB1. HSPB1 knockdown attenuates SHED efficacy and is not fully compensated by PARK7 overexpression. HSPB1 overexpression partially restores SHED-mediated protection under PARK7 silencing.
Neuropathic pain (NP) is chronic pain caused by injury or disease affecting the nervous system. SIN@Lip-HA was prepared and characterised for particle size, zeta potential, encapsulation efficiency and morphology. Sprague-Dawley rats with spared nerve injury (SNI) were randomly divided into the six groups. Analgesic effects were assessed via behavioural tests. Histopathological changes were examined using H E staining. Astrocyte polarisation was detected using immunofluorescence, and Western blotting was used to assess related protein expression. qRT-PCR measured mRNA level of genes, while ELISA evaluated inflammatory cytokines and oxidative stress markers. The study showed that the particle size of SIN@Lip-HA was 111 ± 5.13 nm, with an encapsulation efficiency of 85.47
Oxidative stress, mitochondrial dysfunction, inflammation-associated cellular responses, and apoptosis are closely associated with the pathogenesis of neurodegenerative disorders. Therefore, identifying small molecules capable of modulating these interconnected cellular processes is important for the development of potential neuroprotective strategies. The present study investigated the protective effects of the imidazole derivative (IMD)-1 against hydrogen peroxide (H2O2)-induced oxidative injury in SH-SY5Y human neuroblastoma cells using integrated in vitro and in silico approaches. IMD-1 exhibited strong radical scavenging activity in the 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, reaching 96.8
Globally, acute ischemic stroke (AIS) continues to be a major contributor to death and long-term functional impairment. The present work investigates the clinical relevance of miR-369-3p in AIS and how it regulates brain microvascular endothelial cells. Quantitative measurements of serum miR-369-3p were conducted in a cohort comprising 138 individuals with AIS and 120 healthy controls. ROC curve analysis and Cox proportional hazards regression were employed to evaluate diagnostic and prognostic performance of miR-369-3p. Human brain microvascular endothelial cells (hCMEC/D3) were exposed to oxygen-glucose deprivation followed by reoxygenation (OGD/R), allowing assessment of how miR-369-3p influences cell viability, inflammatory responses, and the expression of adhesion molecules. Downstream target genes were identified through bioinformatics analysis and validated using a dual-luciferase assay. A statistically significant reduction in serum miR-369-3p levels was observed among AIS patients relative to controls (P < 0.001), and this miRNA demonstrated favorable diagnostic accuracy. Lower expression of miR-369-3p emerged as an independent predictor of poorer functional outcomes, as assessed by the modified Rankin Scale (mRS) score at 90 days post‑stroke. In cellular studies, upregulation of miR-369-3p mitigated the loss of cell viability induced by OGD/R, reduced the production of pro‑inflammatory cytokines and lowered the levels of ICAM‑1 and VCAM‑1. Furthermore, VAV3 was confirmed as a direct target of miR-369-3p; restoring VAV3 expression counteracted the protective effects conferred by miR-369-3p. Serum miR-369-3p may serve as a non-invasive diagnostic and prognostic biomarker for AIS. Mechanistically, miR-369-3p protects against ischemic endothelial injury by targeting VAV3 and mitigating inflammation and adhesion molecule expression.
The endocannabinoid system (ECS) plays a key role in regulating neurogenesis and inflammatory processes in the brain. The increasing prevalence of Cannabis use among women highlights the importance of understanding sex-specific effects of cannabinoids, particularly in the context of hormonal interactions. This study aimed to investigate the effects of delta-9-tetrahydrocannabinol (THC) and estradiol benzoate (EB) on adult hippocampal neurogenesis (AHN) and inflammation in ovariectomized female Wistar rats. Sixteen rats were allocated to four experimental groups receiving THC, EB, both treatments, and vehicle. Immunohistochemical analyses were conducted to evaluate markers of proliferation (Ki-67), neurogenesis (doublecortin and PSA-NCAM), cannabinoid receptor expression (CB1), and inflammation (COX-2 and TNF-α) in the hippocampal formation. The administration of THC significantly increased Ki-67 immunoreactivity, suggesting enhanced cell proliferation. A trend toward increased doublecortin expression was observed, particularly in EB-treated animals. THC also modulated CB1 receptor expression, with significant increases in the dentate gyrus and hilus following combined THC and EB treatment. Furthermore, THC reduced inflammatory markers, with region-dependent decreases in COX-2 and TNF-α expression. These findings indicate that THC influences markers associated with hippocampal cell proliferation, neurogenesis, cannabinoid signaling and inflammation in female rats, and that some of these effects depend on estradiol status. The interaction between cannabinoids and gonadal hormones may represent an important mechanism underlying sex-specific neurobiological responses and suggests potential targets for therapeutic intervention in neuropsychiatric disorders.
Activation of microglia and inflammatory response play a central role in the pathological process following intracerebral hemorrhage (ICH). In this study, single-nucleus transcriptomic and proteomic profiling was performed to investigate microglial changes after ICH. A significant upregulation of complement receptor C5aR1 was identified, particularly enriched in microglial subsets exhibiting a pro-inflammatory phenotype. The cellular localization and temporal expression dynamics of C5aR1 were validated by immunofluorescence staining, Western blotting, and quantitative PCR, demonstrating its sustained elevation in microglia after ICH. Pharmacological inhibition with the selective C5aR1 antagonist PMX205 markedly reduced the release of inflammatory cytokines, alleviated neuronal damage, and improved neurological function. Proteomic profiling and subsequent validation suggested that CCR5 may be associated with C5aR1-related inflammatory responses after ICH. Collectively, these findings suggest that C5aR1 serves as a regulatory factor in microglia-mediated neuroinflammation following ICH, and that targeting this pathway may offer a promising therapeutic strategy for hemorrhagic stroke.
The selection of appropriate in vitro inflammatory models is critical for mechanistic studies of neuroinflammation and the screening of anti-inflammatory therapeutics. Lipopolysaccharide (LPS) combined with interferon-γ (IFN-γ) and LPS combined with adenosine triphosphate (ATP) are widely used to induce microglial inflammation; however, their distinct phenotypic characteristics and optimal applications have not been systematically compared. In this study, we combined transcriptomic profiling with functional assays to compare these two inflammatory paradigms in BV2 microglia. Our results showed that both models activated the chemokine–cytokine inflammatory network and induced the production of inflammatory mediators and chemokines, but exhibited distinct molecular and functional biases. LPS/IFN-γ stimulation preferentially upregulated genes such as Lcn2, Ccl2, Upp1, and Pim2, which are associated with innate immune activation, chemokine-mediated inflammatory responses, and cell survival signaling, and was accompanied by marked increases in NO and TNF-α production. In contrast, the LPS/ATP model preferentially upregulated genes associated with ECM/cell adhesion, inflammatory and cellular homeostatic regulation, oxidative stress, and calcium-dependent secretion, including Thbs1, Thbd, Srxn1, Syt7, and Procr. Functionally, this model exhibited more pronounced IL-1β maturation and release, relatively lower NO production, and greater mitochondrial dysfunction and cell injury. Pharmacological inhibition of NLRP3 with MCC950 significantly attenuated ATP-induced IL-1β maturation, supporting the involvement of the NLRP3 inflammasome in the LPS/ATP model. Overall, the LPS/IFN-γ model is more suitable for investigating the NO/iNOS–TNF-α axis, IFN-γ-mediated synergistic pro-inflammatory responses, and classical pro-inflammatory activation, as well as for screening anti-inflammatory agents. In contrast, the LPS/ATP model is more appropriate for studying IL-1β maturation and release, NLRP3 inflammasome activation, mitochondrial damage, and inflammation-related cell death, as well as for screening inflammasome inhibitors. Our findings provide an empirical framework for rational model selection in microglial inflammation research.
Carboxypeptidase E (CPE) may be a therapeutic target for depression. This study investigated the mechanistic insights into the possible regulatory role of CPE in ferroptosis in a mouse model of chronic unpredictable mild stress (CUMS)-induced depression. One week before CUMS exposure, mice received injections of adeno-associated virus-CPE. One week after CUMS induction, the Nrf2 inhibitor ML385 was administered. Behavioral tests were performed 3 weeks after CUMS induction. Ionized calcium-binding adapter molecule 1-positive cells and inflammatory cytokine levels in the CA1 region were assessed by immunofluorescence and ELISA. Hippocampal pathological and neuronal damage were evaluated using H&E and Nissl staining. Additionally, brain-derived neurotrophic factor (BDNF)/nuclear factor erythroid 2-related factor 2 (Nrf2) signaling, ferroptosis markers, oxidative stress indices, glutathione peroxidase 4 (GPX4), and Fe2+ levels were measured using western blot, RT-qPCR, and biochemical assays. CPE overexpression alleviated CUMS-induced depression-like behaviors and hippocampal damage in mice. CPE overexpression reduced microglial activation, neuronal degeneration, and pro-inflammatory factors. It also attenuated oxidative stress by decreasing malondialdehyde, reactive oxygen species, and lactate dehydrogenase while elevating superoxide dismutase and glutathione-S-transferase activities. Mechanistically, CPE activated the BDNF/Nrf2 axis, increased GPX4 activity, suppressed acyl-CoA synthetase long chain family member 4 expression, and reduced iron accumulation. All protective effects of CPE overexpression were reversed by ML385. CPE overexpression activates the BDNF/Nrf2 signaling axis to inhibit ferroptosis, thereby reducing neuroinflammation and depression-like behaviors in mice. This work further clarifies the pathogenesis of depression, enriches pathophysiological theories, and provides key experimental evidence for early intervention and target development.
