Spinal cord injury (SCI) is a prevalent form of spinal cord dysfunction, and the discovery of new effective treatments remains a critical research focus. This study investigated the role of paeoniflorin in a lipopolysaccharide (LPS)-induced PC-12 cell model of SCI and examined the involvement of phosphatase and tensin homolog (PTEN) and the phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) signaling pathway. Rat neuronal PC-12 cells were injured using LPS. Cell viability, proliferation, and apoptosis were assessed using the Cell Counting Kit-8, 5-ethynyl-2′-deoxyuridine (EdU), and Terminal deoxynucleotidyl transferase dUTP Nick End Labeling (TUNEL) assays, respectively. Western blotting and quantitative polymerase chain reaction were used to analyze PI3K, AKT, and PTEN expressions. PTEN was overexpressed to determine its functional role. LPS significantly reduced cell viability and proliferation, while increasing apoptosis. Paeoniflorin treatment ameliorated these injury markers in a dose-dependent manner, downregulated PTEN expression, and enhanced phosphorylated PI3K and AKT levels. PTEN overexpression counteracted the protective effects of paeoniflorin and its activation of PI3K/AKT signaling. Paeoniflorin alleviates LPS-induced cell injury in PC-12 cell model by inhibiting PTEN expression and subsequently activating the PI3K/AKT pathway.

Neuroinflammation and oxidative stress are pivotal drivers of neurological dysfunction following spinal cord injury (SCI). Consequently, precise modulation of pathological glial cells and amelioration of the neuronal microenvironment represent a promising therapeutic strategy. Herein, we developed an injectable, self-healing hydrogel system composed of oxidized sodium alginate, carboxymethyl chitosan, and tannic acid (OCT) for the sustained co-delivery of a Quercetin-Manganese complex (QM) and astrocyte-derived extracellular vesicles encapsulating siRNA (AEVs@siRNA). RNA sequencing revealed significant enrichment of the TNF and chemokine signaling pathways in SCI mice, with a notable upregulation of Serpina3n. This gene, predominantly expressed in astrocytes, modulates their reactive polarization. Leveraging the innate tropism of astrocyte-derived extracellular vesicles, we achieved targeted delivery of Serpina3n-targeting siRNA to astrocytes at the lesion site. This approach effectively suppressed the expression of Serpina3n, inhibiting the transition to a neurotoxic A1 phenotype and alleviating neuronal damage. Concurrently, the sustained release of QM NPs potently scavenged reactive oxygen species, significantly mitigating neuronal ferroptosis. Further mechanistic investigations demonstrated that this combinatorial system attenuated neuroinflammation by inhibiting NF-κB p65 signaling to reduce A1 astrocyte activation, and protected neurons by regulating the SLC7A11/GPX4 axis to counteract apoptosis and ferroptosis, both in vitro and in vivo. Consequently, this targeted delivery system represents a promising approach for enhancing therapeutic efficacy and promoting neural repair following SCI.

Background:Traumatic cervical spinal cord injury (TCSCI) often leads to significant patient paralysis. Current clinical diagnosis relies heavily on empirical interpretation of magnetic resonance imaging (MRI) and the American Spinal Injury Association Impairment Scale (AIS) grade, lacking robust quantitative markers to precisely reflect injury severity. This study aimed to build an artificial intelligence (AI) pipeline for AIS grade prediction based on radiomic features extracted from manually defined regions. Methods:We included 189 patients with TCSCI who underwent MRI within 48 h post-injury. MRI images from 130 patients were used for developing an AI model encompassing image segmentation. Radiomic features were extracted from manually delineated volumes of interest (VOIs). T2-weighted imaging (T2WI) sagittal images were randomly divided into training (n = 104), validation (n = 13), and test (n = 13) sets for segmentation. A total of 183 patients (excluding AIS E) were included in the AIS grade prediction task. Model performance was evaluated using mean dice similarity coefficient (mDICE), mean intersection over union (mIOU), mean specificity, and mean sensitivity. Results:An optimized UCTransnet network, leveraging a Transformer architecture for formal training, combined with a U-Net++ network for pretraining, achieved promising results in segmenting the spinal cord injury site on T2WI sagittal images (mDICE: 0.777 ± 0.021, mIOU: 0.646 ± 0.025, mean specificity: 0.998 ± 0.001, mean sensitivity: 0.895 ± 0.015). Subsequently, an ensemble model (we named Em-En) constructed using selected radiomic features from the manual VOIs demonstrated superior performance for predicting AIS grades in terms of sensitivity, specificity, accuracy, and clinical decision-making benefit compared to other tested models. Conclusions:This study presents an AI-assisted pipeline for predicting the severity of TCSCI. The developed resources provide a theoretical foundation for the clinical application of AI-assisted diagnostic methods, potentially lowering the interpretation barrier for MRI and offering clinicians preliminary quantitative indicators of injury severity. The source code is publicly available.

Introduction:Inflammatory signaling-induced stem cell dysfunction severely impairs bone regeneration. This study aimed to develop a combinatorial strategy using Ti3C2Tx MXene scaffolds and PSAT1-engineered dental pulp stem cells (oe-PSAT1 DPSCs) to counteract inflammation-mediated osteogenic suppression. Methods:Dental pulp stem cells (DPSCs) were treated with TNF-α to simulate an inflammatory microenvironment. miR-665 expression and its targeting relationship with PSAT1 were analyzed via qRT-PCR, dual-luciferase reporter assay, and Western blot. The role of the miR-665/PSAT1/GSK-3β/β-catenin axis in osteogenic differentiation was evaluated using ALP activity, alizarin red staining, and immunofluorescence. Ti3C2Tx MXene was synthesized and characterized, and its effects on ROS scavenging and osteogenesis were assessed in vitro. In vivo efficacy was validated using a rat calvarial defect model with micro-CT, histological staining, and immunohistochemistry. Results:TNF-α stimulation upregulated miR-665, which directly targeted PSAT1 and inhibited the GSK-3β/β-catenin pathway, suppressing DPSCs osteogenic differentiation. PSAT1 overexpression rescued this suppression. Ti3C2Tx MXene scavenged ROS, enhanced calcium-dependent mineralization, and synergized with oe-PSAT1 DPSCs to amplify β-catenin activation. In rat models, the Ti3C2Tx MXene /oe-PSAT1 DPSCs combination achieved superior bone defect closure (higher BV/TV, Tb. Th, and mature collagen deposition) compared to Ti3C2Tx MXene alone. Discussion:This study identifies the miR-665/PSAT1/GSK-3β/β-catenin axis as a key regulator of inflammatory osteogenesis. The Ti3C2Tx MXene/oe-PSAT1 DPSCs strategy concurrently neutralizes oxidative stress and activates osteogenic signaling, providing a translatable platform for inflammatory bone regeneration.

Background Abnormal expression of Zinc finger (ZNF) genes is commonly observed in osteosarcoma (OS), the most prevalent malignant bone tumor in children and teenagers. This project focused on the role of ZNF560 in the progress of OS. Methods The published datasets including TCGA-SARC and GSE99671 was utilized to screen out the abnormal expression of ZNF560 and associated gene patterns in sarcoma and OS tissues. Prognosis value of ZNF560 was identified in TCGA-SARC and OS cohorts. In order to manipulate ZNF560 expression in HOS and MG63 osteosarcoma (OS) cells, genetic strategies such as shRNA constructs were utilized. The expression patterns of ZNF560 were analyzed through techniques such as immunohistochemistry, Western blotting, and qRT-PCR. Results By analyzing data from both the GEO and the Cancer Genome Atlas (TCGA) databases, increased expression of ZNF560 in OS tissues was verified, which was significantly associated with poorer outcomes in osteosarcoma patients both in TCGA-SARC and our own OS cohorts. Additionally, downregulation of ZNF560 resulted in decreased cell viability, fewer colonies, and induced apoptosis of osteosarcoma cells. Moreover, ZNF560 was found to be essential for migration of human osteosarcoma HOS and MG63 cells. Conclusion Collectively, these findings suggest that ZNF560 has the potential to serve as a predictive biomarker for osteosarcoma.

Background: Spinal cord injury (SCI) constitutes a profoundly debilitating neurological disorder precipitating motor and sensory function impairment. Curtailing microglia-driven neuroinflammation alongside oxidative stress proves indispensable for efficacious SCI patient management. Poliumoside (POL), a phenylethanoid glycoside molecule, manifests anti-inflammatory, antioxidant, and neuroprotective capacities. Nevertheless, documentation concerning its SCI therapeutic efficacy remains sparse. Methods: Systemic drug toxicity for two POL dosages (15 mg/kg, 30 mg/kg) was evaluated across multiple organs. An SCI murine model was generated employing Allen’s technique. Mice received random assignment into sham, SCI, and SCI+POL cohorts. Intraperitoneal POL administration ensued for 7 consecutive days post-trauma. Histological staining probed tissue and cellular alterations. Functional recuperation was assessed via the Basso Mouse Scale (BMS), hindlimb flexion scoring, and footprint examination. RNA sequencing (RNA-seq) explored POL’s therapeutic impact within SCI. Immunofluorescence detected the axonal marker neurofilament 200 (NF200), myelin marker myelin basic protein (MBP), and the glial scar indicators ionized calcium-binding adapter molecule 1 and glial fibrillary acidic protein (IBA1, GFAP); Western blot (WB) identified the nerve growth-associated protein 43 (GAP43). WB and immunofluorescence quantified inflammatory and oxidative stress markers. POL’s regulatory function within the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) cascade was scrutinized both in vivo and in vitro. Results: POL intervention induced no systemic organ toxicity. POL-treated mice exhibited pronounced locomotor function enhancement, diminished neuronal tissue depletion, elevated neuronal survival, and attenuated demyelination. RNA-seq analysis illuminated POL’s SCI therapeutic mechanism linkage to axonal regeneration, the phosphatidylinositol signaling apparatus, and the neuronal framework. POL concurrently attenuated glial scar formation and potentiated axonal and myelin regeneration. Mechanistically, POL suppressed pro-inflammatory cytokines and oxidative stress mediators while activating the PI3K/AKT/mTOR pathway. Conclusions: POL mitigated murine spinal cord injury-induced neuroinflammation and oxidative stress through PI3K/AKT/mTOR signaling pathway activation. Furthermore, POL treatment contracted the glial scar expanse within the injury epicenter and fostered axonal regeneration coupled with myelin regeneration. Consequently, POL enhances post-SCI motor function and accelerates neural function restoration.

Activating transcription factor 3 (ATF3) may function as a regulator of various diseases; however, its role in spinal cord injury (SCI) remains unknown. We designed a current work to evaluate the potentials of the ATF3/forkhead box protein A2 (FOXA2) axis in SCI. GSE45006 chip was analyzed, and a volcano plot and heatmap were drawn. Gene Ontology and KEGG analysis were performed for the differentially expressed genes. Animals with SCI were established. Quantitative reverse transcription polymerase chain reaction and western blotting were used to determine mRNA expression, and western blotting was used for detecting protein expression. The interaction between FOXA2 and the ATF3 promoter was evaluated using the UCSC database and confirmed using dual-luciferase and chromatin immunoprecipitation assays. Cellular behaviors were determined using CCK-8, EdU, and TUNEL assays. Levels of p-PI3K, PI3K, p-AKT, and AKT were examined by the WB method. We found that ATF3 expression was markedly increased in rats with SCI. Interestingly, ATF3 knockdown increased the proliferation and suppressed the apoptotic ability of PC12 cells. FOXA2 activates ATF3 transcription. Knockdown of FOXA2-mediated down-regulation of ATF3 increases growth and decreases PC12 cell death. ATF3 knockdown could increase the level of p-PI3K and p-AKT; FOXA2 shRNA could affect the expression of p-PI3K and p-AKT, which was partially abrogated by ATF3 OE. Forkhead box protein regulates the transcription of ATF3, thereby affecting cell growth and PC12 cell death.

Reversing the decline of microglia in the dentate gyrus of stressed animals has antidepressant effects, but the molecular mechanisms are unclear. Since microglia normally interact with astrocytes and astrocytic purinergic 2Y1 receptor (P2Y1R) signaling plays an important role in regulating cellular crosstalk, we hypothesize that astrocytic P2Y1R signaling may mediate the antidepressant effects of microglia stimulation. Our results showed that a single injection of low-dose lipopolysaccharide (LPS) (100 μg/kg) elicited rapid antidepressant effects and a significant increase in adenosine triphosphate (ATP) levels in the dentate gyrus in chronically stressed mice, and that these effects of LPS were abolished by chemogenetic inhibition of microglia. Depletion of endogenous ATP, non-specific antagonization of purinergic receptors, or specific inhibition of P2Y1Rs, but not other purinergic receptors, by MRS2179 in the hippocampus abolished the antidepressant effects of low-dose LPS. Conditional gene knockout data showed that the antidepressant effect of low-dose LPS could not be observed in mice lacking P2Y1Rs in astrocytes but not in forebrain neurons. Chemogenetic inhibition of microglia in the dentate gyrus, specific deletion of P2Y1Rs in astrocytes and the absence of ATP abolished the increase in doublecortin (DCX)+ cells and brain-derived neurotrophic factor (BDNF) induced by a low dose of LPS in the dentate gyrus of stressed mice, and infusion of BDNF antibodies into the hippocampus simultaneously abolished the pro-neurogenesis and antidepressant effects of microglia stimulation in stressed mice. Taken together, these results suggest that ATP signaling mobilized by microglia stimulation has an antidepressant effect by triggering astrocytic P2Y1R-dependent synthesis of BDNF.

Rationale: Necroptosis in astrocytes induced by mitochondrial dysfunction following spinal cord injury (SCI) significantly contributes to neuronal functional deficits. Mitophagy plays a crucial role in clearing damaged mitochondria and inhibiting necroptosis. Fanconi anemia complementation group C (FANCC), a member of the Fanconi anemia gene family, exerts a protective role by facilitating mitophagy in immune processes. However, the role of FANCC in SCI-induced astrocytic necroptosis and the underlying mechanisms remain unexplored. Methods: Astrocyte-specific FANCC conditional knockout (Fanccfl/fl-GFAP-Cre) mice, obtained by mating Fanccfl/fl mice with GFAP-Cre mice, served as a model of moderate thoracic spinal cord contusion injuries. Using bulk and single-nucleus RNA sequencing, we investigated the protective role of FANCC in astrocytes after SCI. We assessed necroptosis and mitophagy in astrocytes through quantitative PCR, western blotting, flow cytometry, immunofluorescence, and transmission electron microscopy. Molecular mechanisms were explored via co-immunoprecipitation, proteomics, molecular docking, and confocal imaging. Computer virtual screening identified poliumoside as a FANCC activator. Histopathological staining and functional assessments (gait analysis, Basso Mouse Scale, and hindlimb reflex score) were conducted to evaluate the therapeutic effects of poliumoside on SCI. Results: Astrocytic FANCC deficiency exacerbated necroptosis and mitochondrial damage, leading to severe neurological deficits. Conversely, FANCC overexpression increased PTEN-induced kinase 1-Parkin expression, thereby activating mitophagy and reducing necroptosis. Proteomics revealed FANCC's interaction with a specific peptide of TANK-binding kinase 1 (TBK1), which further promoted mitophagy. Treatment with the FANCC activator poliumoside improved neural pathology and motor function recovery in SCI mice. Conclusion: The current study indicated that FANCC interacts with TBK1 and consequently mediates Parkin translocation, activates mitophagy, and inhibits astrocyte necroptosis. Our findings demonstrate the neuroprotective role and therapeutic potential of FANCC for SCI amelioration.

Objectives:Laminoplasty (LAMP) is a common procedure for multilevel cervical spondylotic myelopathy (MCSM). The traditional K-line is a guide for LAMP candidate selection but is inferior to the modified K-line (mK-line) in predicting clinical outcomes. The spinal cord line (SC-line) is another indicator that considers anterior compression but is not typically used for selecting surgical segments. This study intended to propose and validate the combined application of modified spinal cord line (mSC-line) with mK-line for surgical decision-making in MCSM patients. Methods:This study included 63 MCSM patients categorized into K-line(-) group and K-line(+) group, or Type I group and Type II group based on SC-line. We defined mK-line and mSC-line in sagittal T2WI MRIs. All patients with both mK-line(+) and mSC-line(+) underwent standard LAMP. Radiographic analysis was conducted using CCI, mK-INT and mSC-INT. Clinical outcomes were evaluated by JOA, NDI and VAS scores. Preoperative and postoperative radiological outcomes and clinical outcomes were used to evaluate the prognosis and the efficacy of segmental decision-making. Results:There were no difference in baseline characteristics among all the participants. Post-operative spinal cord shift indicators (mK-INT and mSC-INT) increased significantly. The JOA score increased, while NDI and VAS scores decreased. Both the radiological outcomes and clinical outcomes demonstrated a good prognosis even in K-line(-) group and Type II group. There was a statistical correlation between JOA score recovery rate with both mK-INT and mSC-INT. Conclusions:The presence of mK-line(+) and mSC-line(+) in MRI is crucial for the selection of surgical segments in LAMP for MCSM patients. This combined criterion can help predict sufficient decompression of the cervical spinal cord and good clinical outcomes.

Spinal cord injury (SCI) is a devastating condition characterized by the accumulation of myelin debris (MD), persistent neuroinflammation, and impaired neural regeneration. Although macrophages are pivotal for MD clearance, the impact of excessive MD phagocytosis on macrophage phenotype and function remains poorly understood. Building upon our prior evidence that exendin-4 (Ex-4), a glucagon-like peptide-1 receptor (GLP-1R) agonist, mitigates microglia-driven neuroinflammation post-SCI, this study elucidates the therapeutic efficacy and underlying mechanisms of Ex-4 in alleviating macrophage senescence, restoring efferocytotic capacity, and facilitating neural repair. Employing a T10 contusive SCI model in male C57BL/6 mice, in vivo administration of Ex-4 was combined with macrophage-specific knockdown of growth arrest-specific 6 (Gas6) via AAV-shRNA. Complementary in vitro assays involved bone marrow-derived macrophages (BMDMs) challenged with MD in the presence or absence of Ex-4 or AMP-activated protein kinase (AMPK) inhibition. Cellular senescence and efferocytosis were comprehensively assessed through live-cell imaging, immunofluorescence, senescence-associated β-galactosidase staining, quantitative PCR, and western blotting. Molecular docking and dynamics simulations elucidated GLP-1R–AMPK interactions, corroborated by in vivo validation. Results demonstrate that MD-engulfing macrophages exhibit foam cell-like morphology and upregulated senescence markers, including increased β-galactosidase activity and senescence-associated secretory phenotype, concomitant with diminished efferocytosis via downregulation of the Axl receptor. Senescent macrophages were shown to exacerbate neuronal apoptosis and astrocytic scar formation in co-culture systems. Ex-4 treatment significantly attenuated macrophage senescence, restored efferocytotic function, and reduced neuronal injury and astrocyte activation, effects contingent upon AMPK/Gas6/Axl pathway activation and abrogated by Gas6 knockdown. In vivo, Ex-4 administration enhanced remyelination, axonal regeneration, and functional recovery, while attenuating glial scar formation following SCI. Collectively, these findings identify macrophage senescence induced by excessive MD phagocytosis as a novel pathological contributor to SCI progression and establish Ex-4 as a promising therapeutic agent that restores macrophage homeostasis and promotes neural repair via GLP-1R/AMPK/Gas6/Axl signaling.

BackgroundSpinal cord injury (SCI) is a debilitating condition within the nervous system with a high disability rate and substantial economic burden. The functional recovery following SCI is enhanced by moderate levels of autophagy but hindered when autophagy becomes excessive. Galectin-3 (GAL3) has been recognized as an autophagy regulator; however, its role in SCI and its associated mechanism are largely unknown.MethodsThe Walsh clamping method was employed to establish a rat SCI model, while a high-concentration glutamate incubation method was used to create an in vitro model of spinal cord neuronal injury. Subsequent to establishing the injury models, the expression levels of GAL3 were detected using QPCR and Western Blot. Immunohistochemical staining was performed to determine the localization of GAL3 expression. SiR-GAL3 or GAL3 inhibitors were utilized to knock down or inhibit GAL3 expression, and behavioral analysis was conducted to assess the recovery of motor function in rats following SCI. Bioinformatics analysis was carried out to explore the mechanism of action of GAL3 post-SCI. Western Blot was used to examine the relationship between the expression levels of GAL3 and autophagy-related proteins following SCI. Sequencing analysis was performed to identify the differential gene expression in spinal cord neurons with knocked-down GAL3 compared to the control group after neural injury, aiming to investigate the mechanism of action between GAL3 and its downstream target gene Cell-division-cycle-42 (CDC42). Co-IP was employed to detect the interaction between GAL3 and CDC42 proteins. Western Blot was used to analyze the relationship between CDC42 and autophagy-related protein expression levels following in vitro stimulation of neurons with GAL3. Molecular biology experiments were conducted to assess the expression levels and localization of CDC42 post-SCI. Behavioral analysis was performed to evaluate the recovery of motor function in rats with inhibited CDC42 expression after SCI. ELISA was used to measure the expression levels of GAL3 and CDC42 in both rat and human samples post-SCI.ResultsWe found that GAL3 was increased in spinal neurons and serum in SCI rats, and knockdown or inhibition of GAL3 promoted motor function recovery. The bioinformatics analysis showed that GAL3 is closely related to programmed cell death after SCI. Indeed, the knockdown of GAL3 resulted in a decrease in autophagy markers ATG7 and LC3 II/I ratio, along with an increase in P62 expression. Furthermore, GAL3 and CDC42 exhibited close associations with neuronal autophagy. Injection of siR-CDC42 and CDC42 inhibitor ML141 effectively reduced GAL3-mediated enhancement of neuronal autophagy. Additionally, CDC42 was increased in spinal neurons post-SCI, and administration of ML141 decreased the expression of autophagy markers and improved motor function recovery. Importantly, elevated levels of GAL3 and CDC42 were observed in the serum of SCI patients.

Spinal cord injury (SCI) is a severe neurological complication following spinal fracture, which has long posed a challenge for clinicians. Microglia play a dual role in the pathophysiological process after SCI, both beneficial and detrimental. The underlying mechanisms of microglial actions following SCI require further exploration. The present study combined three different machine learning algorithms, namely weighted gene co-expression network analysis, random forest analysis and least absolute shrinkage and selection operator analysis, to screen for differentially expressed genes in the GSE96055 microglia dataset after SCI. It then used protein-protein interaction networks and gene set enrichment analysis with single genes to investigate the key genes and signaling pathways involved in microglial function following SCI. The results indicated that microglia not only participate in neuroinflammation but also serve a significant role in the clearance mechanism of apoptotic cells following SCI. Notably, bioinformatics analysis and lipopolysaccharide + UNC569 (a MerTK-specific inhibitor) stimulation of BV2 cell experiments showed that the expression levels of Anxa2, Myo1e and Spp1 in microglia were significantly upregulated following SCI, thus potentially involved in regulating the clearance mechanism of apoptotic cells. The present study suggested that Anxa2, Myo1e and Spp1 may serve as potential targets for the future treatment of SCI and provided a theoretical basis for the development of new methods and drugs for treating SCI.

Excitotoxicity represents the primary cause of neuronal death following spinal cord injury (SCI). While autophagy plays a critical and intricate role in SCI, the specific mechanism underlying the relationship between excitotoxicity and autophagy in SCI has been largely overlooked. In this study, we isolated primary spinal cord neurons from neonatal rats and induced excitotoxic neuronal injury by high concentrations of glutamic acid, mimicking an excitotoxic injury model. Subsequently, we performed transcriptome sequencing. Leveraging machine learning algorithms, including weighted correlation network analysis (WGCNA), random forest analysis (RF), and least absolute shrinkage and selection operator analysis (LASSO), we conducted a comprehensive investigation into key genes associated with spinal cord neuron injury. We also utilized protein-protein interaction network (PPI) analysis to identify pivotal proteins regulating key gene expression and analyzed key genes from public datasets (GSE2599, GSE20907, GSE45006, and GSE174549). Our findings revealed that six genes—Anxa2, S100a10, Ccng1, Timp1, Hspb1, and Lgals3—were significantly upregulated not only in vitro in neurons subjected to excitotoxic injury but also in rats with subacute SCI. Furthermore, Hspb1 and Lgals3 were closely linked to neuronal autophagy induced by excitotoxicity. Our findings contribute to a better understanding of excitotoxicity and autophagy, offering potential targets and a theoretical foundation for SCI diagnosis and treatment.

Spinal cord injury (SCI) patients have an increased susceptibility to coronary heart disease (CHD) due to dysregulated lipid deposition. We conducted a comprehensive investigation to gain insights into the specific roles of Apolipoprotein B-100 (APOB-100) in the development of CHD in patients suffering from SCI. First, we established an SCI rat model through semitransection. APOB-100 expression in plasma exosomes obtained from patients were determined. Subsequently, we found APOB-100 affected macrophage polarization when treating co-cultured neurons/macrophages lacking Sortilin with extracellular vesicles derived from SCI rats, where APOB-100 co-immunoprecipitated with Sortilin. Moreover, APOB-100 upregulation reduced neuronal cell viability and triggered apoptosis by upregulating Sortilin, leading to a decline in the Basso, Beattie, and Bresnahan (BBB) scale, exacerbation of neuron injury, increased macrophage infiltration, and elevated blood lipid-related indicators in SCI rats, which could be reversed by silencing Sortilin. In conclusion, APOB-100 from post-SCI patients' extracellular vesicles upregulates Sortilin, thereby endangering those patients to CHD.

Facet joint osteoarthritis (FJOA), a condition commonly observed in individuals of middle to old age, has been relatively under-researched compared to other subtypes of osteoarthritis (OA). This study investigated the role of transcription factor FoxO1 in FJOA using a Col2a1-creERT knock-in mouse model. It was found that FoxO1 deletion led to severe osteoarthritic changes, indicating that FoxO1 played a critical role in cartilage homeostasis. Transcriptome sequencing was performed on degenerated cartilage from FoxO1-deleted mice. This process identified differentially expressed genes (DEGs), offering insights into the molecular mechanisms underlying FJOA. Bioinformatics analysis, including Kyoto Encyclopedia of Genes and Genomes (KEGG), Gene Set Enrichment Analysis (GSEA) and protein-protein interaction (PPI) network analysis, identified Itgb3, Itga1, Itga6, Itga7, Itga8, Itga10, Col1a1, and Il6, as potential key contributors to FJOA after FoxO1 deletion. Importantly, overexpression of Itgb3 and inhibition of Il6 counteracted FoxO1 knockdown-induced impairments in chondrocyte migration and extracellular matrix synthesis, respectively. This study discovered FoxO1 as a key regulator of the pathogenesis of FJOA, helped unravel the complex molecular mechanisms underlying FJOA, and contributed to the development of promising therapeutic avenues toward FJOA.

Background The treatment of spinal cord injury (SCI) has always been a significant research focus of clinical neuroscience, with inhibition of microglia-mediated neuro-inflammation as well as oxidative stress key to successful SCI patient treatment. Caffeic acid phenethyl ester (CAPE), a compound extracted from propolis, has both anti-inflammatory and anti-oxidative effects, but its SCI therapeutic effects have rarely been reported. Methods We constructed a mouse spinal cord contusion model and administered CAPE intraperitoneally for 7 consecutive days after injury, and methylprednisolone (MP) was used as a positive control. Hematoxylin–eosin, Nissl, and Luxol Fast Blue staining were used to assess the effect of CAPE on the structures of nervous tissue after SCI. Basso Mouse Scale scores and footprint analysis were used to explore the effect of CAPE on the recovery of motor function by SCI mice. Western blot analysis and immunofluorescence staining assessed levels of inflammatory mediators and oxidative stress-related proteins both in vivo and in vitro after CAPE treatment. Further, reactive oxygen species (ROS) within the cytoplasm were detected using an ROS kit. Changes in mitochondrial membrane potential after CAPE treatment were detected with 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide. Mechanistically, western blot analysis and immunofluorescence staining were used to examine the effect of CAPE on the SIRT1/PGC1α/DRP1 signaling pathway. Results CAPE-treated SCI mice showed less neuronal tissue loss, more neuronal survival, and reduced demyelination. Interestingly, SCI mice treated with CAPE showed better recovery of motor function. CAPE treatment reduced the expression of inflammatory and oxidative mediators, including iNOS, COX-2, TNF-α, IL-1β, 1L-6, NOX-2, and NOX-4, as well as the positive control MP both in vitro and in vivo. In addition, molecular docking experiments showed that CAPE had a high affinity for SIRT1, and that CAPE treatment significantly activated SIRT1 and PGC1α, with down-regulation of DRP1. Further, CAPE treatment significantly reduced the level of ROS in cellular cytoplasm and increased the mitochondrial membrane potential, which improved normal mitochondrial function. After administering the SIRT1 inhibitor nicotinamide, the effect of CAPE on neuro-inflammation and oxidative stress was reversed.On the contrary, SIRT1 agonist SRT2183 further enhanced the anti-inflammatory and antioxidant effects of CAPE, indicating that the anti-inflammatory and anti-oxidative stress effects of CAPE after SCI were dependent on SIRT1. Conclusion CAPE inhibits microglia-mediated neuro-inflammation and oxidative stress and supports mitochondrial function by regulating the SIRT1/PGC1α/DRP1 signaling pathway after SCI. These effects demonstrate that CAPE reduces nerve tissue damage. Therefore, CAPE is a potential drug for the treatment of SCI through production of anti-inflammatory and anti-oxidative stress effects. Graphical Abstract

The objective of this study was to analyze differentially expressed genes (DEGs) in rats with spinal cord injury (SCI) undergoing treatment with menstrual blood-derived stem cells (MenSCs) and treadmill training, compared to untreated SCI rats. Spinal cord injury models were induced in SD rats via dorsal hemisection at T10, following which rats were divided into CTMT (cells and treadmill training) and SCI groups. In the CTMT group, MenSCs (1.0×10 5 ) were microinjected into the injured area, followed by 2 weeks of aerobic exercise training. Transcriptome sequencing was performed on injured spinal cord tissues, with subsequent analyses encompassing differential gene expression, GO pathway enrichment, KEGG pathway enrichment, and protein-protein interaction (PPI) network analysis. Motor function recovery was assessed using the Basso, Bresnahan, and Beattie (BBB) scoring system, while local pathological changes were evaluated via H&E staining. Validation of DEGs was conducted using qRT-PCR and Western blot. Transcriptome analysis revealed 247 up-regulated and 174 down-regulated genes in the CTMT group, with notable up-regulated genes including Bdnf, Hmox1, Sd4, Mmp3, and Cd163. KEGG and GO analyses indicated involvement in growth, development, metabolism, and immune-inflammatory processes, with the PI3K-Akt signaling pathway enriched for BDNF genes. CTMT led to significant improvements in locomotor function and local pathology, accompanied by increased BDNF gene expression. The combined approach of MenSCs and treadmill training effectively enhanced motor function recovery and upregulated BDNF gene expression in SCI rats, suggesting a promising intervention target for SCI treatment and introducing a novel avenue for SCI management.

BACKGROUND:Spinal cord injury (SCI) is a devastating disease that can lead to tissue loss and neurological dysfunction. TNIP2 is a negative regulator of NF-κB signaling due to its capacity to bind A20 and suppress inflammatory cytokines-induced NF-κB activation. However, the anti-inflammatory role of TNIP2 in SCI remains unclear. Our study's intention was to evaluate the effect of TNIP2 on the inflammatory response of microglia after spinal cord injury in rats.METHODS:HE staining and Nissl staining were performed on day 3 following SCI to analyze the histological changes. To further investigate the functional changes of TNIP2 after SCI, we performed immunofluorescence staining experiments. The effect of LPS on TNIP2 expression in BV2 cells was examined by western blot. The levels of TNF-α, IL-1β, and IL-6 in spinal cord tissues of rats with SCI and in BV2 cells with LPS were measured by using qPCR.RESULTS:TNIP2 expression was closely associated with the pathophysiology of SCI in rats, and TNIP2 was involved in regulating functional changes in microglia. TNIP2 expression was increased during SCI in rats and that overexpression of TNIP2 inhibited M1 polarization and pro-inflammatory cytokine production in microglia, which might ultimately protect against inflammatory responses through the MAPK and NF-κB signaling pathways.CONCLUSIONS:The present study provides evidence for a role of TNIP2 in the regulation of inflammation in SCI and suggests that induction of TNIP2 expression alleviated the inflammatory response of microglia.