Biochemistry and Biophysics Reports最新文献

{"title":"Altered lipidomic and metabolomic status in cerebrospinal fluid of children with myelin oligodendrocyte glycoprotein antibody-associated disorder","authors":"Pin Fee Chong ,&nbsp;Kenta Kajiwara ,&nbsp;Yuji Ueno ,&nbsp;Satoshi Akamine ,&nbsp;Hiroyuki Torisu ,&nbsp;Ryutaro Kira ,&nbsp;Shouichi Ohga ,&nbsp;Yasunari Sakai","doi":"10.1016/j.bbrep.2025.102233","DOIUrl":"10.1016/j.bbrep.2025.102233","url":null,"abstract":"<div><div>Myelin oligodendrocyte glycoprotein antibody-associated disorder (MOGAD) is a group of acquired demyelinating syndromes affecting the central nervous system. MOGAD-associated bioactive molecules remain elusive. A retrospective case-control study was performed to characterize the biochemical and immunological profiles of cerebrospinal fluid (CSF) in MOGAD. Thirteen patients with MOGAD (onset age: 2–14 years, 6 females) and five patients with epilepsy, serving as controls, were enrolled. Liquid chromatography with tandem mass spectrometry was used for lipidomic and metabolomic analyses using CSF samples collected at disease onset (n = 5). The MS/MS system detected a total of 7527 molecules in lipidomic and 17,526 molecules in metabolomic analyses of CSF. Among them, 162 (0.02 %) lipophilic molecules were detected at levels that differed from those of controls. Among the 549 (0.03 %) hydrophilic molecules that were differentially presented, pyridoxine, ribitol, and isethionate levels were significantly lower in patients with MOGAD. Both lipidomic and metabolomic analyses, discriminated CSF samples of patients with MOGAD from controls using Uniform Manifold Approximation and Projection. In summary, CSF samples from children with MOGAD exhibit distinctive lipidomic and metabolomic profiles. These findings provide evidence for the diagnostic potential of CSF-based lipidomic and metabolomic analyses for childhood-onset MOGAD.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102233"},"PeriodicalIF":2.2,"publicationDate":"2025-09-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144933273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of bile powder based on component data matrix and progressive matching strategy combined with UHPLC-QTOF-MS","authors":"Haonan Wu ,&nbsp;Xianrui Wang ,&nbsp;Minghua Li ,&nbsp;Xiaohan Guo ,&nbsp;Xianlong Cheng ,&nbsp;Tian Yin ,&nbsp;Wenguang Jing ,&nbsp;Feng Wei","doi":"10.1016/j.bbrep.2025.102234","DOIUrl":"10.1016/j.bbrep.2025.102234","url":null,"abstract":"<div><h3>Background</h3><div>Pig bile powder (PBP), chicken bile powder (CKBP), goat bile powder (GBP), bear bile powder (BBP), cow bile powder (CBP), rabbit bile powder (RBP) and snake bile powder (SBP) are commonly used bile powder medicinal materials in the clinic. Due to frequent confusion and fraudulent use, it is necessary to establish and enrich identification methods. In this paper, the component data matrix (CDM) contained common component matrix (CCM) and specificity component matrix (SCM), and progressive matching strategy (PMS) were innovatively proposed to identify PBP, CKBP, GBP, BBP, CBP, RBP and SBP.</div></div><div><h3>Methods</h3><div>The samples were analyzed by LC-MS to obtain MS data on chemical components. Then, the CCM and SCM of each bile powder was obtained by taking the intersection and de-intersection of MS data. Finally, the CCM and SCM were used for matching test samples to obtain matching value (MV). At the same time, the proprietary components were further explored.</div></div><div><h3>Results</h3><div>Based on CDM and PMS, the identification of PBP, CKBP, GBP, BBP, CBP, RBP and SBP can be quickly realized with MV ≥ 70 %. <strong>Conclusions</strong>: The CDM and PMS were efficient to identify PBP, CKBP, GBP, BBP, CBP, RBP and SBP, which has more specificity. It can enrich identification methods and strengthen quality assurance of bile powder medicinal materials.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102234"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144926003","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"CEBPB as a prognostic biomarker and its association with immune cells in clear cell renal cell carcinoma","authors":"Yaoqiang Ren ,&nbsp;Min Wei ,&nbsp;Quanfa Tian ,&nbsp;Wenke Guo","doi":"10.1016/j.bbrep.2025.102231","DOIUrl":"10.1016/j.bbrep.2025.102231","url":null,"abstract":"<div><div>Clear cell renal cell carcinoma (ccRCC) is a highly aggressive malignancy with a poor prognosis. This study examines the expression, prognostic significance, and immune association of CCAAT/enhancer-binding protein beta (CEBPB) in ccRCC. RNA sequencing data from The Cancer Genome Atlas (TCGA) and Genotype-Tissue Expression (GTEx) Project were analyzed using the STAR workflow and R software. Immunohistochemistry (IHC) validated CEBPB expression in ccRCC tissues. Functional enrichment analyses (Gene Ontology and Kyoto Encyclopedia of Genes and Genomes) and a protein-protein interaction (PPI) network (STRING and Cytoscape) were used to explore CEBPB-related pathways. Single-sample gene set enrichment analysis (ssGSEA) revealed significant correlations between CEBPB expression and the infiltration of 24 immune cell types. CEBPB was linked to immune-related pathways, including humoral immune response, leukocyte migration, and cytokine signaling. PPI analysis identified strong interactions with STAT3/EP300, highlighting its role in immune regulation. Cox regression analysis showed that high CEBPB expression is associated with poorer overall survival, supporting its potential as a prognostic biomarker. In conclusion, CEBPB plays a key role in shaping the immune microenvironment of ccRCC and may serve as a novel prognostic marker and therapeutic target.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102231"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144922047","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A modified protocol for the isolation and culture of human umbilical vein endothelial cells","authors":"Anmol Sandhu ,&nbsp;Anannya Parvathi ,&nbsp;Jennifer Jane McGhee ,&nbsp;Salim Ismail ,&nbsp;I-Ping Loh ,&nbsp;Bert van der Werf ,&nbsp;Jie Zhang ,&nbsp;Trevor Sherwin","doi":"10.1016/j.bbrep.2025.102221","DOIUrl":"10.1016/j.bbrep.2025.102221","url":null,"abstract":"<div><div>The umbilical cord is a valuable source of foetal stem cells, progenitor cells, and early-stage developmental cells, including human umbilical vein endothelial cells (HUVECs). HUVECs are widely used as a model for endothelial biology and are increasingly being investigated for their regenerative potential. Efficient isolation of these cells from the umbilical vein is a critical first step for both research and therapeutic applications. To date, most published protocols utilise Collagenase A for isolation. In this study, we present a modified HUVEC isolation protocol that employs dispase, alongside refined tissue and cell culture handling practices. We characterised the isolated cells using established HUVEC markers CD31 and CD146, and demonstrated in situ detachment of the cells from the vessel wall through immunofluorescence imaging. Our method achieved a success rate exceeding 95.6 % across all umbilical cords processed. These findings highlight the protocol's potential for broad applicability across research settings, using readily accessible reagents and equipment.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102221"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144922054","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Clinical application study of a novel fully automatic erythrocyte osmotic fragility analysis system","authors":"Zixin Deng ,&nbsp;Yi Li ,&nbsp;Zhizhi Xiang ,&nbsp;Yi Liu ,&nbsp;Jingbo Tang ,&nbsp;Song Zhang ,&nbsp;Guangchao Zang ,&nbsp;Yingying Gao ,&nbsp;Lei Ma","doi":"10.1016/j.bbrep.2025.102224","DOIUrl":"10.1016/j.bbrep.2025.102224","url":null,"abstract":"<div><h3>Background</h3><div>Existing erythrocyte osmotic fragility test (EFT) methods are constrained by subjectivity, poor reproducibility, and lack of standardization, limiting their clinical utility in thalassemia screening. This study evaluates the RA-800 Plus, a novel fully automated EFT analyzer, as a scalable tool for thalassemia screening and aims to establish reference intervals for healthy individuals. By addressing the key limitations of manual EFTs, this work seeks to promote methodological innovation and enable standardized, high-throughput screening in diverse clinical settings.</div></div><div><h3>Methods</h3><div>EFTs were performed on 273 healthy adults via an RA-800 Plus analyzer. The light scattering turbidity method was employed to establish a reference interval within the normal range. The neonatal samples were tested in the same way to determine the newborn-specific range. Moreover, 97 samples underwent dual tests of RA-800 Plus analysis and the gold standard (genetic testing) to evaluate the diagnostic performance. A total of 103 pairs of samples were detected and compared via an automated system and the traditional manual direct colorimetric method, further verifying the consistency of the two methods. The effects of anticoagulant type, sample storage, and rewarming were also investigated.</div></div><div><h3>Results</h3><div>RA-800 Plus demonstrated high reliability, with minimal influence from anticoagulants and sample handling conditions. The analyzer showed an 84 % detection rate for β-thalassemia, indicating superior effectiveness for β-thalassemia screening. The test results were consistent with the manual method (Kappa ≥0.6). ROC curve analysis confirmed the suitability of both methods for thalassemia screening, with AUCs of 0.91 for β-thalassemia and 0.72 for α-thalassemia.</div></div><div><h3>Conclusions</h3><div>The RA-800 Plus offers a fully automated, reliable, and clinically viable alternative for thalassemia screening, particularly for β-thalassemia. Its stability, accuracy, and ease of use make it a valuable tool for improving thalassemia diagnostics.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102224"},"PeriodicalIF":2.2,"publicationDate":"2025-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144926005","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Collagen proteins, thrombospondin 1 and lumican are differentially expressed across breast cancer subtypes by functional proteomics from core needle biopsy samples of Taiwanese breast cancer","authors":"Wei-Chi Ku ,&nbsp;Nam Nhut Phan ,&nbsp;Chih-Yi Liu ,&nbsp;Chi-Jung Huang ,&nbsp;Chen-Chung Liao ,&nbsp;Yen-Chun Huang ,&nbsp;Po-Hsin Kong ,&nbsp;Ling-Ming Tseng ,&nbsp;Chi-Cheng Huang","doi":"10.1016/j.bbrep.2025.102210","DOIUrl":"10.1016/j.bbrep.2025.102210","url":null,"abstract":"<div><h3>Purpose</h3><div>This study aimed to conduct functional proteomics across breast cancer subtypes with bioinformatics analyses.</div></div><div><h3>Methods</h3><div>Candidate proteins were identified using nanoscale liquid chromatography with tandem mass spectrometry (NanoLC-MS/MS) from core needle biopsy samples of early stage (0-III) breast cancers, followed by external validation with public domain gene-expression datasets (TCGA TARGET GTEx and TCGA BRCA).</div></div><div><h3>Results</h3><div>Seventeen proteins demonstrated significantly differential expression and protein-protein interaction (PPI) found the strong networks including COL2A1, COL11A1, COL6A1, COL6A2, THBS1 and LUM. Public domain databases also showed that <em>COL2A1</em>, <em>COL11A1</em>, <em>COL6A1</em>, <em>COL6A2</em> and <em>LUM</em> were higher in primary/metastatic tumor than in normal tissue (one-way ANOVA, all P-values less than 0.001), and all six genes were differentially expressed across four molecular subtypes based on hormone receptor (HR) status and human epidermal growth factor receptor II (HER2) status (one-way ANOVA, all P-values less than 0.001). Disease-specific survival discrepancy was observed comparing breast cancer patients of the upper and lower quartile of the collagen family (<em>COL2A1</em>, <em>COL11A1</em>, <em>COL6A1</em>, <em>COL6A2</em>), <em>THBS1</em> and <em>LUM</em> gene expression signature (log-rank test, P = 0.06).</div></div><div><h3>Conclusion</h3><div>Functional proteomics suggested that collagen proteins, thrombospondin 1 and lumican are differentially expressed across breast cancer subtypes.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102210"},"PeriodicalIF":2.2,"publicationDate":"2025-08-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144919914","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Comparative analysis of RT-qPCR, flow cytometry, and Di-4-ANEPPDHQ fluorescence for distinguishing macrophages phenotypes","authors":"Ahmed Abu Siniyeh ,&nbsp;Walhan Alshaer ,&nbsp;Nirmeen Elzogheir ,&nbsp;Majed Al-Holi ,&nbsp;Dana A. Alqudah ,&nbsp;Duaa Abuarqoub ,&nbsp;Joanna M. Kwiatek","doi":"10.1016/j.bbrep.2025.102225","DOIUrl":"10.1016/j.bbrep.2025.102225","url":null,"abstract":"<div><div>This study evaluates the effectiveness of fluorescence microscopy using Di-4-ANEPPDHQ in differentiating macrophage phenotypes (M0, M1, and M2) compared to RT-qPCR and flow cytometry. Using THP-1 monocyte-derived macrophages, we assessed cytokine expression (IL-1β, IL-6, IL-10) via RT-qPCR, surface markers (CD86, CD64, CD206) through flow cytometry, and membrane properties with Di-4-ANEPPDHQ fluorescence. RT-qPCR showed significant differences in cytokine expression: M1 macrophages had elevated IL-1β (p &lt; 0.0001) and IL-6 (p &lt; 0.0001), while M2 macrophages exhibited higher IL-10 levels (p = 0.0030). Flow cytometry revealed distinct surface marker profiles, with M1 expressing high CD64 and M2 showing increased CD206. Di-4-ANEPPDHQ fluorescence indicated membrane order differences: M1 macrophages were depolarized (red shift), while M2 macrophages were hyperpolarized (blue shift). Statistical analysis confirmed high sensitivity and specificity for RT-qPCR and flow cytometry, while Di-4-ANEPPDHQ fluorescence technique provides real-time observations of changes in macrophage membrane behavior, enhancing understanding of their dynamic properties under various conditions. These findings highlight the value of integrating these methods for comprehensive macrophage phenotype characterization, which can aid in understanding macrophage polarization in immune responses and disease contexts.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102225"},"PeriodicalIF":2.2,"publicationDate":"2025-08-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144917091","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The multifaceted role of SQSTM1/p62 in disc degeneration: A master regulator of cellular stress responses","authors":"Yang Hou ,&nbsp;Lei Liu ,&nbsp;Tianyi Zhao,&nbsp;Yongfei Guo ,&nbsp;Jiangang Shi","doi":"10.1016/j.bbrep.2025.102222","DOIUrl":"10.1016/j.bbrep.2025.102222","url":null,"abstract":"<div><div>Intervertebral disc degeneration (IDD) is a key contributor to lumbar degenerative diseases and chronic low back pain. Accumulating evidence indicates that Sequestosome 1 (SQSTM1/p62), a multifunctional adaptor protein, plays a pivotal role in IDD pathogenesis through its regulation of autophagy, oxidative stress, inflammation, and programmed cell death. This review summarizes the multifaceted functions of SQSTM1 in the context of IDD, including its involvement in the autophagy-lysosome pathway, antioxidant defense via the Keap1-Nrf2 axis, activation of the NF-κB signaling and NLRP3 inflammasome, and modulation of apoptosis, pyroptosis, and ferroptosis. Moreover, SQSTM1 contributes to extracellular matrix degradation by upregulating matrix metalloproteinases and downregulating their inhibitors. Given its dynamic expression during disc degeneration, SQSTM1 holds promise as both a biomarker for IDD progression and a therapeutic target. Potential strategies targeting SQSTM1 include the use of autophagy inducers, inflammatory pathway inhibitors, and ferroptosis/pyroptosis modulators. However, challenges remain in precisely modulating SQSTM1 activity and translating findings into clinical therapies. Future research leveraging advanced technologies such as single-cell RNA sequencing, proteomics, and organoid models is essential to unravel the complex, stage- and cell-specific roles of SQSTM1 in IDD. Understanding these mechanisms may open new avenues for effective treatment and improved patient outcomes in degenerative spinal disorders.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102222"},"PeriodicalIF":2.2,"publicationDate":"2025-08-29","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144913809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring the biomechanical complexity of glioblastoma spheroids and organoids with co-localized Brillouin and Raman microspectroscopy","authors":"Roberta Galli ,&nbsp;Jan Rix ,&nbsp;Tina Leonidou ,&nbsp;Katrin Kirsche ,&nbsp;Edmund Koch ,&nbsp;Achim Temme ,&nbsp;Ilker Y. Eyüpoglu ,&nbsp;Ortrud Uckermann","doi":"10.1016/j.bbrep.2025.102227","DOIUrl":"10.1016/j.bbrep.2025.102227","url":null,"abstract":"<div><div>Brillouin microscopy allows mechanical investigations of biological materials at the subcellular level and can be integrated with Raman spectroscopy for simultaneous chemical mapping, thus enabling a more comprehensive interpretation of biomechanics. The present study investigates different in vitro glioblastoma models using a combination of Brillouin and Raman microspectroscopy. Spheroids of the U87-MG cell line and two patient-derived cell lines as well as patient-derived organoids were used. Brillouin microscopy provided maps of viscoelastic parameters, while Raman spectroscopy identified key biochemical components such as proteins, lipids, glycogen and cholesterol. Cluster analysis of the Raman spectra allowed the categorization of biochemical groups and the correlation of their Brillouin shift and bandwidth across the different glioblastoma models. The results showed that spheroids from the same cell line exhibited relatively homogeneous biomechanical properties, while differences existed between different cell lines. In contrast, organoids from the same patient exhibited greater mechanical and biochemical heterogeneity. Brillouin shift and bandwidth showed significant variation among Raman clusters, highlighting the need to consider biochemical composition in biomechanical assessments. The cytoplasmic protein cluster was biochemically and biomechanically consistent across models, while lipid- and glycogen-related clusters varied. The approach used in this study facilitates the interpretation of Brillouin data in heterogeneous biological systems and allows comparisons between different models. The results emphasize the need for multimodal analysis for correct interpretation of biomechanical measurements in complex tissues and for comparison between heterogeneous samples.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102227"},"PeriodicalIF":2.2,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144913810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"5-HMF plays an intervention role in ischemic stroke by regulating GluR2 to improve mitochondrial function and promote angiogenesis","authors":"Yan Zhang ,&nbsp;Peipei Yuan ,&nbsp;Yaxin Wei ,&nbsp;Saifei Li ,&nbsp;Lirui Zhao ,&nbsp;Qingyun Ma ,&nbsp;Yiran Huo ,&nbsp;Xiaoke Zheng ,&nbsp;Weisheng Feng","doi":"10.1016/j.bbrep.2025.102208","DOIUrl":"10.1016/j.bbrep.2025.102208","url":null,"abstract":"<div><h3>Introduction</h3><div>5-Hydroxymethyl furfural (5-HMF) is a furan compound with a molecular formula of C₆H₆O₃. Studies have found that 5-HMF has many pharmacological effects, such as improving hemorheology, anti-inflammatory, antioxidant activity and anti-myocardial ischemia. Identifying the preventive effect of 5-HMF against ischemic stroke and its possible mechanism was the aim of this investigation.</div></div><div><h3>Methods</h3><div>The MCAO animal model was used to detect the related indexes of behavior, neurological function and mitochondrial function, and to clarify the effect of 5-HMF on ischemic stroke. The HBMECs model induced by OGD-R was intervened by 5-HMF, and the GluR2 silencing sequence was added to detect mitochondrial function and angiogenesis-related indicators, so as to further explore the mechanism of 5-HMF intervention in ischemic stroke.</div></div><div><h3>Results</h3><div>In the MCAO experiment, the behavioral results showed that 5-HMF increased the quantity of autonomic activities, increased the exercise time and distance, reduced the balance beam score, and prolonged the residence time of MCAO rats on the rotating rod (<em>P</em> &lt; 0.01). The results of neurological function indicated that 5-HMF reduced the neurological function score, increased the blood flow velocity of middle cerebral artery (<em>P</em> &lt; 0.01), reduced the area of cerebral infarction, and alleviated the damage of cerebral cortex and hippocampal neurons. The results of mitochondrial function indicated that 5-HMF could significantly reduce the apoptosis of primary brain cells, ROS, Ca²⁺ levels, increase mitochondrial membrane potential and GluR2 expression in MCAO rats (<em>P</em> &lt; 0.01). In the OGD-R-induced HBMECs experiment, 5-HMF increased the expression level of GluR2, increased cell viability, and decreased LDH activity in cell supernatant (<em>P</em> &lt; 0.01). The results of mitochondrial function showed that 5-HMF reduced the levels of Ca²⁺ and glutamate in the cell supernatant, reduced the levels of apoptosis and ROS, increased the mitochondrial membrane potential, and improved the expression of mitochondrial dynamics-related proteins (<em>P</em> &lt; 0.05 or <em>P</em> &lt; 0.01). These improvements were markedly reversed after the addition of GluR2 silencing sequence (<em>P</em> &lt; 0.01).</div></div><div><h3>Conclusions</h3><div>5-HMF has the effect of ameliorating brain damage in MCAO rats, and by regulating the expression of GluR2, reducing intracellular Ca²⁺ concentration, improving mitochondrial function, promoting angiogenesis, thereby reducing OGD-R-induced HBMECs damage.</div></div>","PeriodicalId":8771,"journal":{"name":"Biochemistry and Biophysics Reports","volume":"44 ","pages":"Article 102208"},"PeriodicalIF":2.2,"publicationDate":"2025-08-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144908061","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}