Pub Date : 2025-10-14DOI: 10.1016/j.biocel.2025.106872
Haser H. Sutcu , Arthur Thomas--Joyeux , Mikaël Cardot-Martin , Delphine Dugué , François Vianna , Yann Perrot , Mohamed A. Benadjaoud , Marc Benderitter , Céline Baldeyron
DNA integrity and stability are vital for proper cellular activity. Nevertheless, to treat cancer patients, DNA is the main target for inducing tumoral cell death. Nowadays, cancer treatment is improving by the development of new technologies, protocols and strategies. Amongst them, the charged particle radiotherapies are becoming prevalent. However, tumor-neighboring healthy tissues are still exposed to ionizing radiation (IR) and subject to late side effects. Skeletal muscle is one of those tissues most likely to be affected. To decipher the DNA damage response (DDR) of skeletal muscle cells, myogenic cells, we irradiated them with microbeams of protons or α-particles and followed the accumulation of DDR proteins at localized irradiation sites. Thereby, we showed that myoblasts, proliferating myogenic cells, repair local IR-induced DNA damage through both non-homologous end-joining and homologous recombination with different recruitment dynamics depending on the characteristics of ionizing particles (type, energy deposition and time after irradiation), whereas myotubes, post-mitotic myogenic cells, display globally reduced DNA damage response.
{"title":"The DNA damage response in myogenic C2C7 cells depends on the characteristics of ionizing particles","authors":"Haser H. Sutcu , Arthur Thomas--Joyeux , Mikaël Cardot-Martin , Delphine Dugué , François Vianna , Yann Perrot , Mohamed A. Benadjaoud , Marc Benderitter , Céline Baldeyron","doi":"10.1016/j.biocel.2025.106872","DOIUrl":"10.1016/j.biocel.2025.106872","url":null,"abstract":"<div><div>DNA integrity and stability are vital for proper cellular activity. Nevertheless, to treat cancer patients, DNA is the main target for inducing tumoral cell death. Nowadays, cancer treatment is improving by the development of new technologies, protocols and strategies. Amongst them, the charged particle radiotherapies are becoming prevalent. However, tumor-neighboring healthy tissues are still exposed to ionizing radiation (IR) and subject to late side effects. Skeletal muscle is one of those tissues most likely to be affected. To decipher the DNA damage response (DDR) of skeletal muscle cells, myogenic cells, we irradiated them with microbeams of protons or α-particles and followed the accumulation of DDR proteins at localized irradiation sites. Thereby, we showed that myoblasts, proliferating myogenic cells, repair local IR-induced DNA damage through both non-homologous end-joining and homologous recombination with different recruitment dynamics depending on the characteristics of ionizing particles (type, energy deposition and time after irradiation), whereas myotubes, post-mitotic myogenic cells, display globally reduced DNA damage response.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106872"},"PeriodicalIF":2.8,"publicationDate":"2025-10-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145309766","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-10-06DOI: 10.1016/j.biocel.2025.106870
Hai-yang Liao , Guo-hua Zhang , Jian-xiong Zheng , Jin-yue Lu , Jia-yao Hao , Min Tan , Zhan-dong Wang , Hai-li Shen
Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by persistent synovial inflammation and progressive joint destruction. Fibroblast-like synovial cells (FLSs) are the main effector cells in the synovial microenvironment that cause chronic swelling and joint injury, and their enhanced glycolytic metabolism can lead to persistent joint injury. As a key regulatory enzyme in glycolysis, pyruvate kinase M2 (PKM2) plays a crucial role in the pathogenesis of RA. However, the exact mechanism by which PKM2 induces the inflammatory response of RA-FLSs through enhanced glucose metabolism and its impact on the pathogenic behaviour of cells remain unclear. This study detected the expression of PKM2 in synovial tissues and RA-FLSs of patients with RA and explored the effect of PKM2 on collagen-induced arthritis (CIA) rats. The results showed that PKM2 was upregulated in the synovial tissue of RA and RA-FLSs. PKM2 could promote glucose uptake, ATP and lactic acid production, and extracellular acidification rate in RA-FLSs, thereby promoting the release of pro-inflammatory cytokines such as TNF-α, interleukin-1 β (IL-1β), and IL-6. However, inhibiting PKM2 can reverse these changes. In in vivo experiments, inhibition of PKM2 could significantly improve the clinical arthritis symptoms of CIA rats (reduce plantar swelling and arthritis score), down-regulate the expression of pro-inflammatory cytokines, and inhibit bone erosion in CIA rats, reducing inflammatory cell infiltration, synovial hyperplasia and joint destruction. Furthermore, inhibiting PKM2 can suppress the phosphorylated expression of Akt and mTOR proteins, thereby inhibiting glycolytic reprogramming. Our research results indicate that PKM2 mediates glycolytic reprogramming to induce the release of RA-FLSs inflammatory cytokines by activating the Akt/mTOR signaling pathway, thereby promoting the progression of RA. Therefore, PKM2 may be a candidate target for the treatment of RA. Targeting PKM2 to regulate glycolytic reprogramming can provide a new idea for the treatment of RA.
{"title":"Pyruvate kinase M2 (PKM2) regulates rheumatoid arthritis by mediating glycolysis reprogramming through the Akt/mTOR pathway","authors":"Hai-yang Liao , Guo-hua Zhang , Jian-xiong Zheng , Jin-yue Lu , Jia-yao Hao , Min Tan , Zhan-dong Wang , Hai-li Shen","doi":"10.1016/j.biocel.2025.106870","DOIUrl":"10.1016/j.biocel.2025.106870","url":null,"abstract":"<div><div>Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by persistent synovial inflammation and progressive joint destruction. Fibroblast-like synovial cells (FLSs) are the main effector cells in the synovial microenvironment that cause chronic swelling and joint injury, and their enhanced glycolytic metabolism can lead to persistent joint injury. As a key regulatory enzyme in glycolysis, pyruvate kinase M2 (PKM2) plays a crucial role in the pathogenesis of RA. However, the exact mechanism by which PKM2 induces the inflammatory response of RA-FLSs through enhanced glucose metabolism and its impact on the pathogenic behaviour of cells remain unclear. This study detected the expression of PKM2 in synovial tissues and RA-FLSs of patients with RA and explored the effect of PKM2 on collagen-induced arthritis (CIA) rats. The results showed that PKM2 was upregulated in the synovial tissue of RA and RA-FLSs. PKM2 could promote glucose uptake, ATP and lactic acid production, and extracellular acidification rate in RA-FLSs, thereby promoting the release of pro-inflammatory cytokines such as TNF-α, interleukin-1 β (IL-1β), and IL-6. However, inhibiting PKM2 can reverse these changes. In in vivo experiments, inhibition of PKM2 could significantly improve the clinical arthritis symptoms of CIA rats (reduce plantar swelling and arthritis score), down-regulate the expression of pro-inflammatory cytokines, and inhibit bone erosion in CIA rats, reducing inflammatory cell infiltration, synovial hyperplasia and joint destruction. Furthermore, inhibiting PKM2 can suppress the phosphorylated expression of Akt and mTOR proteins, thereby inhibiting glycolytic reprogramming. Our research results indicate that PKM2 mediates glycolytic reprogramming to induce the release of RA-FLSs inflammatory cytokines by activating the Akt/mTOR signaling pathway, thereby promoting the progression of RA. Therefore, PKM2 may be a candidate target for the treatment of RA. Targeting PKM2 to regulate glycolytic reprogramming can provide a new idea for the treatment of RA.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106870"},"PeriodicalIF":2.8,"publicationDate":"2025-10-06","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145253430","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
This study aimed to elucidate the regulatory role of the muscle-specific gene ATP1B4 in skeletal muscle metabolism and mitophagy in diabetic sarcopenia (DS) rats.
Methods
Differentially expressed genes were screened from the GEO dataset GSE7014, and ATP1B4 was identified as a candidate gene associated with DS. A DS rat model was established via high-fat diet feeding and streptozotocin injection. ATP1B4 expression was modulated through lentiviral overexpression or knockdown. Additionally, PI3K/AKT/mTOR pathway activators (SC79, leucine) and inhibitors (LY294002, MK-2206) were administered. Protein expression of ATP1B4, phosphorylated PI3K/AKT/mTOR components, and autophagy markers (LC3-II, DRP1, ATG9, MFN2) was assessed via Western blotting, immunohistochemistry, and immunofluorescence. Skeletal muscle function and structure were evaluated using behavioral tests (treadmill and inclined plane) and histopathological staining (H&E, Masson, PAS).
Results
Bioinformatic analysis of the GSE7014 dataset identified ATP1B4 as a skeletal muscle-related differentially expressed gene enriched in extracellular matrix and metabolic pathways. In DS rats, ATP1B4 expression was upregulated, coinciding with suppression of PI3K/AKT/mTOR signaling and activation of mitophagy markers (LC3-II, DRP1, ATG9). Overexpression of ATP1B4 exacerbated hyperglycemia, muscle atrophy, collagen accumulation, and glycogen deposition, while knockdown reversed these effects. Activation of the PI3K/AKT/mTOR pathway improved muscle function and histological architecture, normalized autophagy, and reduced pathological features. However, co-overexpression of ATP1B4 eliminated the protective effects of pathway activation. Conversely, dual intervention with ATP1B4 knockdown and PI3K activation restored skeletal muscle integrity and autophagy flux. Importantly, ATP1B4 expression remained unchanged following pathway modulation, supporting its unidirectional upstream regulatory role in DS.
Conclusion
ATP1B4 may aggravate diabetic sarcopenia by acting as an upstream suppressor of the PI3K/AKT/mTOR pathway.
{"title":"ATP1B4 as a candidate upstream regulator of muscle atrophy in diabetic sarcopenia via PI3K/AKT/mTOR-mediated autophagy","authors":"Tingting Duan , Shumin Jia , Dan Zhou , Liqun Zhao","doi":"10.1016/j.biocel.2025.106869","DOIUrl":"10.1016/j.biocel.2025.106869","url":null,"abstract":"<div><h3>Objective</h3><div>This study aimed to elucidate the regulatory role of the muscle-specific gene ATP1B4 in skeletal muscle metabolism and mitophagy in diabetic sarcopenia (DS) rats.</div></div><div><h3>Methods</h3><div>Differentially expressed genes were screened from the GEO dataset GSE7014, and ATP1B4 was identified as a candidate gene associated with DS. A DS rat model was established via high-fat diet feeding and streptozotocin injection. ATP1B4 expression was modulated through lentiviral overexpression or knockdown. Additionally, PI3K/AKT/mTOR pathway activators (SC79, leucine) and inhibitors (LY294002, MK-2206) were administered. Protein expression of ATP1B4, phosphorylated PI3K/AKT/mTOR components, and autophagy markers (LC3-II, DRP1, ATG9, MFN2) was assessed via Western blotting, immunohistochemistry, and immunofluorescence. Skeletal muscle function and structure were evaluated using behavioral tests (treadmill and inclined plane) and histopathological staining (H&E, Masson, PAS).</div></div><div><h3>Results</h3><div>Bioinformatic analysis of the GSE7014 dataset identified ATP1B4 as a skeletal muscle-related differentially expressed gene enriched in extracellular matrix and metabolic pathways. In DS rats, ATP1B4 expression was upregulated, coinciding with suppression of PI3K/AKT/mTOR signaling and activation of mitophagy markers (LC3-II, DRP1, ATG9). Overexpression of ATP1B4 exacerbated hyperglycemia, muscle atrophy, collagen accumulation, and glycogen deposition, while knockdown reversed these effects. Activation of the PI3K/AKT/mTOR pathway improved muscle function and histological architecture, normalized autophagy, and reduced pathological features. However, co-overexpression of ATP1B4 eliminated the protective effects of pathway activation. Conversely, dual intervention with ATP1B4 knockdown and PI3K activation restored skeletal muscle integrity and autophagy flux. Importantly, ATP1B4 expression remained unchanged following pathway modulation, supporting its unidirectional upstream regulatory role in DS.</div></div><div><h3>Conclusion</h3><div>ATP1B4 may aggravate diabetic sarcopenia by acting as an upstream suppressor of the PI3K/AKT/mTOR pathway.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106869"},"PeriodicalIF":2.8,"publicationDate":"2025-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145193857","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
β-Tricalcium phosphate (β-TCP), frequently employed for bone tissue regeneration, can induce inflammation during the initial phases of implantation within the organism. However, mechanisms by which β-TCP nanoparticles (NPs) cause this inflammatory response is rarely reported. This project aims to investigate the causes of the macrophage inflammatory response induced by β-TCP NPs. Here, macrophage-like RAW264.7 cells were co-cultured with conditioned medium containing β-TCP NPs to identify the pathways through which β-TCP NPs influence inflammation and polarization of macrophages. This effect is achieved by modulating mitochondrial oxidative stress in the immune microenvironment. The results demonstrated that β-TCP NPs caused mitochondrial swelling, increased intracellular calcium ions, reduced mitochondrial membrane potential, as well as decreased the level of adenosine triphosphate (ATP) and translocase of outer mitochondrial membrane 20 (TOMM20). These NPs further lead to mitochondrial oxidative damage. These alterations promoted the polarization of macrophage to M1-type. Exogenous mitochondrial-targeted antioxidants could block this M1-type macrophage polarization. The findings of this work suggest that β-TCP NPs induce macrophage inflammation and contribute to M1 macrophage polarization, primarily through the activation of mitochondrial oxidative stress. These insights could guide the development of improved β-TCP formulations to mitigate inflammatory responses in bone regeneration applications.
{"title":"β-Tricalcium phosphate nanoparticles induce macrophage polarization to M1-type through mitochondrial oxidative stress activation","authors":"Yunbing Chen , Xinping Li , Hongyi Yang, Gaoying Ran, Lifang Zhang, Shuguang Zeng","doi":"10.1016/j.biocel.2025.106868","DOIUrl":"10.1016/j.biocel.2025.106868","url":null,"abstract":"<div><div>β-Tricalcium phosphate (β-TCP), frequently employed for bone tissue regeneration, can induce inflammation during the initial phases of implantation within the organism. However, mechanisms by which β-TCP nanoparticles (NPs) cause this inflammatory response is rarely reported. This project aims to investigate the causes of the macrophage inflammatory response induced by β-TCP NPs. Here, macrophage-like RAW264.7 cells were co-cultured with conditioned medium containing β-TCP NPs to identify the pathways through which β-TCP NPs influence inflammation and polarization of macrophages. This effect is achieved by modulating mitochondrial oxidative stress in the immune microenvironment. The results demonstrated that β-TCP NPs caused mitochondrial swelling, increased intracellular calcium ions, reduced mitochondrial membrane potential, as well as decreased the level of adenosine triphosphate (ATP) and translocase of outer mitochondrial membrane 20 (TOMM20). These NPs further lead to mitochondrial oxidative damage. These alterations promoted the polarization of macrophage to M1-type. Exogenous mitochondrial-targeted antioxidants could block this M1-type macrophage polarization. The findings of this work suggest that β-TCP NPs induce macrophage inflammation and contribute to M1 macrophage polarization, primarily through the activation of mitochondrial oxidative stress. These insights could guide the development of improved β-TCP formulations to mitigate inflammatory responses in bone regeneration applications.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106868"},"PeriodicalIF":2.8,"publicationDate":"2025-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145158799","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-23DOI: 10.1016/j.biocel.2025.106867
Linshan Xie , Rong Xu , Huining Liu , Man Na , Qikai Qin , Fei Xu , Raymond C. Stevens , Yan Liu
The biased agonism of glucagon-like peptide-1 receptor (GLP-1R) plays a key role in the efficacy and side effects of drugs used to treat type II diabetes mellitus and obesity. Despite its therapeutic potential, the mechanisms underlying GLP-1R biased agonism remain poorly understood. In this study, we investigate the role of the extracellular domain (ECD) in GLP-1R signaling bias through saturation mutagenesis at seven key sites. We examined 126 mutations and identified several that selectively abolished β-arrestin recruitment while retaining cAMP production. Additionally, we employed a large language model (LLM) to interpret the functional impacts of these mutations, uncovering correlations between sequence features and signaling outcome. These findings provide new insight into the "two-domain" model of class B1 G protein-coupled receptors (GPCRs), highlighting the ECD's role in biased agonism and offering novel information for designing more effective and selective GLP-1R agonists.
{"title":"Characterizing the role of extracellular domain in GLP-1R biased agonism","authors":"Linshan Xie , Rong Xu , Huining Liu , Man Na , Qikai Qin , Fei Xu , Raymond C. Stevens , Yan Liu","doi":"10.1016/j.biocel.2025.106867","DOIUrl":"10.1016/j.biocel.2025.106867","url":null,"abstract":"<div><div>The biased agonism of glucagon-like peptide-1 receptor (GLP-1R) plays a key role in the efficacy and side effects of drugs used to treat type II diabetes mellitus and obesity. Despite its therapeutic potential, the mechanisms underlying GLP-1R biased agonism remain poorly understood. In this study, we investigate the role of the extracellular domain (ECD) in GLP-1R signaling bias through saturation mutagenesis at seven key sites. We examined 126 mutations and identified several that selectively abolished β-arrestin recruitment while retaining cAMP production. Additionally, we employed a large language model (LLM) to interpret the functional impacts of these mutations, uncovering correlations between sequence features and signaling outcome. These findings provide new insight into the \"two-domain\" model of class B1 G protein-coupled receptors (GPCRs), highlighting the ECD's role in biased agonism and offering novel information for designing more effective and selective GLP-1R agonists.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106867"},"PeriodicalIF":2.8,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145151893","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-23DOI: 10.1016/j.biocel.2025.106866
Yongwei Du , Fangtian Xu , Shengwang Miao , Bo Zhou , Jianwen Mo , Zhen Liu
<div><h3>Background</h3><div>Ferroptosis occurs in osteoblasts in a diabetic environment, which impairs osteoblast number and function, promotes osteoblast death, destroys bone homeostasis, and eventually contributes to type 2 diabetic osteoporosis (T2DOP). Chromobox protein homolog 7 (CBX7) deficiency plays a positive role in bone formation and skeletal development. Besides, CBX7 interference has been reported to protect against disease development by inhibiting ferroptosis. This study focuses on determining whether CBX7 is involved in the progression of T2DOP by regulating osteoblast ferroptosis and explore the underlying mechanism.</div></div><div><h3>Methods</h3><div>The bone tissues of 28 patients with T2DOP (the research group) and severe bone trauma (the control group) were collected, and CBX7 expression in bone tissues was detected through RT-qPCR and western blotting. Mouse pre-osteoblast MC3T3-E1 cells were treated with high glucose (HG; 25 mM) for 72 h to establish an <em>in vitro</em> model of T2DOP. The effects of CBX7 knockdown or overexpression on ferroptosis in HG-treated MC3T3-E1 cells were assessed by examining the levels of intracellular Fe<sup>2 +</sup> , lipid peroxidation, ROS, MDA, 4‑HNE, GSH, and ferroptosis regulatory proteins SLC7A11 and GPX4. The osteogenic differentiation of HG-treated MC3T3-E1 cells after knocking down or overexpressing CBX7 was assessed by performing ALP and ARS staining and measuring the levels of osteogenesis markers (ALP, RUNX2, and OCN). Whether CBX7 knockdown affects osteoblast differentiation and ferroptosis by regulating the Nrf2 pathway was validated by using the Nrf2 inhibitor ML385. Furthermore, Sprague Dawley (SD) rats were fed a high-fat diet and injected with streptozotocin to induce type 2 diabetes (T2DM), followed by the induction of T2DOP for 2 months and subsequently the determination of the anti-osteoporotic, anti-ferroptotic, and pro-osteogenic effects of CBX7 deficiency <em>in vivo</em>.</div></div><div><h3>Results</h3><div>CBX7 expression was markedly increased in the bone tissues of T2DOP patients compared to control patients. HG stimulation enhanced CBX7 expression, intracellular Fe<sup>2+</sup>, lipid peroxidation, ROS, MDA, and 4‑HNE levels but attenuated GSH, SLC7A11, and GPX4 levels in MC3T3-E1 cells. These effects of HG were reversed by CBX7 knockdown but were intensified by CBX7 overexpression. CBX7 silencing antagonized while CBX7 overexpression enhanced the inhibitory effects of HG treatment on the osteogenic differentiation of MC3T3-E1 cells. CBX7 knockdown promoted Nrf2 expression and nuclear translocation in HG-treated MC3T3-E1 cells and CBX7 showed potential protein interaction with Nrf2. The inhibition of CBX7 depletion on HG-induced ferroptosis in MC3T3-E1 cells and its promotion on the osteogenic differentiation of MC3T3-E1 cells were abrogated by ML385. Additionally, CBX7 silencing ameliorated bone mass loss and bone microstructure destruction, reduced MDA and i
{"title":"Chromobox protein homolog 7 (CBX7) deficiency inhibits osteoblast ferroptosis by activating the Nrf2 function in type 2 diabetic osteoporosis","authors":"Yongwei Du , Fangtian Xu , Shengwang Miao , Bo Zhou , Jianwen Mo , Zhen Liu","doi":"10.1016/j.biocel.2025.106866","DOIUrl":"10.1016/j.biocel.2025.106866","url":null,"abstract":"<div><h3>Background</h3><div>Ferroptosis occurs in osteoblasts in a diabetic environment, which impairs osteoblast number and function, promotes osteoblast death, destroys bone homeostasis, and eventually contributes to type 2 diabetic osteoporosis (T2DOP). Chromobox protein homolog 7 (CBX7) deficiency plays a positive role in bone formation and skeletal development. Besides, CBX7 interference has been reported to protect against disease development by inhibiting ferroptosis. This study focuses on determining whether CBX7 is involved in the progression of T2DOP by regulating osteoblast ferroptosis and explore the underlying mechanism.</div></div><div><h3>Methods</h3><div>The bone tissues of 28 patients with T2DOP (the research group) and severe bone trauma (the control group) were collected, and CBX7 expression in bone tissues was detected through RT-qPCR and western blotting. Mouse pre-osteoblast MC3T3-E1 cells were treated with high glucose (HG; 25 mM) for 72 h to establish an <em>in vitro</em> model of T2DOP. The effects of CBX7 knockdown or overexpression on ferroptosis in HG-treated MC3T3-E1 cells were assessed by examining the levels of intracellular Fe<sup>2 +</sup> , lipid peroxidation, ROS, MDA, 4‑HNE, GSH, and ferroptosis regulatory proteins SLC7A11 and GPX4. The osteogenic differentiation of HG-treated MC3T3-E1 cells after knocking down or overexpressing CBX7 was assessed by performing ALP and ARS staining and measuring the levels of osteogenesis markers (ALP, RUNX2, and OCN). Whether CBX7 knockdown affects osteoblast differentiation and ferroptosis by regulating the Nrf2 pathway was validated by using the Nrf2 inhibitor ML385. Furthermore, Sprague Dawley (SD) rats were fed a high-fat diet and injected with streptozotocin to induce type 2 diabetes (T2DM), followed by the induction of T2DOP for 2 months and subsequently the determination of the anti-osteoporotic, anti-ferroptotic, and pro-osteogenic effects of CBX7 deficiency <em>in vivo</em>.</div></div><div><h3>Results</h3><div>CBX7 expression was markedly increased in the bone tissues of T2DOP patients compared to control patients. HG stimulation enhanced CBX7 expression, intracellular Fe<sup>2+</sup>, lipid peroxidation, ROS, MDA, and 4‑HNE levels but attenuated GSH, SLC7A11, and GPX4 levels in MC3T3-E1 cells. These effects of HG were reversed by CBX7 knockdown but were intensified by CBX7 overexpression. CBX7 silencing antagonized while CBX7 overexpression enhanced the inhibitory effects of HG treatment on the osteogenic differentiation of MC3T3-E1 cells. CBX7 knockdown promoted Nrf2 expression and nuclear translocation in HG-treated MC3T3-E1 cells and CBX7 showed potential protein interaction with Nrf2. The inhibition of CBX7 depletion on HG-induced ferroptosis in MC3T3-E1 cells and its promotion on the osteogenic differentiation of MC3T3-E1 cells were abrogated by ML385. Additionally, CBX7 silencing ameliorated bone mass loss and bone microstructure destruction, reduced MDA and i","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106866"},"PeriodicalIF":2.8,"publicationDate":"2025-09-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145151910","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-22DOI: 10.1016/j.biocel.2025.106865
Zhenxing Liu , Jingye Zhao , Lei Zhang , Xiaoting Wu , Jiamiao Liu , Yuanrui Mei , Shuyan Liu , Jieru Lin , Hongyan Li , Xiaoye Qi , Fuping Lu , Huabing Zhao , Aipo Diao
Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by impaired skin barrier function and immune dysregulation. Autophagy, a lysosome-dependent degradation pathway essential for removing unnecessary components, plays a crucial role in maintaining cellular homeostasis. Defective autophagy has been implicated in AD pathogenesis, and enhancing autophagic activity represents a viable therapeutic strategy. This study investigated the potential of the natural saponin escin to ameliorate AD through autophagy activation. We demonstrated that escin induced autophagy in HaCaT keratinocytes and mitigated tight junction (TJ) barrier disruption in an AD-like cell model stimulated with IL-4 and IL-13. Notably, silencing ATG7, an essential autophagy-related protein, abrogated the barrier-restorative effects of escin. Furthermore, in a 2,4-dinitrochlorobenzene (DNCB)-induced murine model of AD, escin treatment ameliorated AD-like skin lesions, reduced mast cell infiltration, and decreased cutaneous levels of the pro-inflammatory cytokines IL-4, IL-13, and IFN-γ. Escin administration also restored the epidermal expression of key TJ proteins, Claudin-1 and ZO-1. Mechanistically, escin promoted the nuclear translocation of transcription factor EB (TFEB) and upregulated the expression of genes involved in autophagy and lysosome biogenesis. These protective effects were associated with the activation of the AMPK-mTORC1-TFEB signaling pathway. Collectively, our findings indicate that escin enhances autophagy and restores skin barrier function, highlighting its potential as a novel therapeutic agent for AD treatment.
{"title":"Escin alleviates DNCB-induced atopic dermatitis-like symptoms by promoting autophagy activation and tight junction barrier restoration","authors":"Zhenxing Liu , Jingye Zhao , Lei Zhang , Xiaoting Wu , Jiamiao Liu , Yuanrui Mei , Shuyan Liu , Jieru Lin , Hongyan Li , Xiaoye Qi , Fuping Lu , Huabing Zhao , Aipo Diao","doi":"10.1016/j.biocel.2025.106865","DOIUrl":"10.1016/j.biocel.2025.106865","url":null,"abstract":"<div><div>Atopic dermatitis (AD) is a chronic inflammatory skin disease characterized by impaired skin barrier function and immune dysregulation. Autophagy, a lysosome-dependent degradation pathway essential for removing unnecessary components, plays a crucial role in maintaining cellular homeostasis. Defective autophagy has been implicated in AD pathogenesis, and enhancing autophagic activity represents a viable therapeutic strategy. This study investigated the potential of the natural saponin escin to ameliorate AD through autophagy activation. We demonstrated that escin induced autophagy in HaCaT keratinocytes and mitigated tight junction (TJ) barrier disruption in an AD-like cell model stimulated with IL-4 and IL-13. Notably, silencing ATG7, an essential autophagy-related protein, abrogated the barrier-restorative effects of escin. Furthermore, in a 2,4-dinitrochlorobenzene (DNCB)-induced murine model of AD, escin treatment ameliorated AD-like skin lesions, reduced mast cell infiltration, and decreased cutaneous levels of the pro-inflammatory cytokines IL-4, IL-13, and IFN-γ. Escin administration also restored the epidermal expression of key TJ proteins, Claudin-1 and ZO-1. Mechanistically, escin promoted the nuclear translocation of transcription factor EB (TFEB) and upregulated the expression of genes involved in autophagy and lysosome biogenesis. These protective effects were associated with the activation of the AMPK-mTORC1-TFEB signaling pathway. Collectively, our findings indicate that escin enhances autophagy and restores skin barrier function, highlighting its potential as a novel therapeutic agent for AD treatment.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106865"},"PeriodicalIF":2.8,"publicationDate":"2025-09-22","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145139104","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-19DOI: 10.1016/j.biocel.2025.106864
Chloé Richard , Charlène Pourpe , Guillaume Fourneaux , Gwendal Cueff , Laurent Parry , Cécile Coudy-Gandilhon , Jonas Kindberg , Alina L. Evans , Andrea Miller , Guillemette Gauquelin-Koch , Christophe Tatout , Cécile Polge , Daniel Taillandier , Fabrice Bertile , Etienne Lefai , Lydie Combaret
Muscle atrophy is observed in several pathophysiological situations, including physical inactivity, leading to negative health consequences, without any effective treatment currently available. Conversely, brown bears resist muscle atrophy during hibernation, despite prolonged physical inactivity and fasting. We previously reported that hibernating brown bear serum increases protein content in human myotubes and inhibits proteolysis. To go further, we deciphered here the transcriptional effects of brown bear serum in human myotubes using large-scale transcriptomics. After 48 h, the winter-hibernating bear serum (WBS) induced a specific transcriptomic program, affecting mostly biological pathways related to muscle growth and BMP signalling, compared to the summer-active bear (SBS) serum. WBS predominantly reduced, at mRNA and protein levels, activators and inhibitors of BMP signalling, which is associated with muscle mass maintenance. Moreover, BMP activity was more responsive to a stimulation by BMP7 at supra-physiological concentrations in human myotubes cultured in WBS versus SBS conditions. Meanwhile, WBS also up-regulated expression of genes encoding repressors of the pro-atrophic TGF-β pathway, decreased phosphorylated SMAD3 nuclear protein levels, and down-regulated TGF-β target genes. Furthermore, WBS treatment resulted in reduced TGF-β signalling responsiveness in human myotubes stimulated with TGF-β3 at physiological concentrations. Overall, even though WBS induced larger transcriptomic changes in the BMP compared to TGF-β pathway, the functional consequences were more pronounced for the TGF-β pathway with a marked inhibition. This study suggests that bioactive compounds in WBS may protect human muscle cells during catabolic situations, by regulating the TGF-β/BMP balance. These findings open new perspectives for therapies targeting muscle atrophy.
{"title":"Hibernating brown bear serum modulates the balance of TGF-β and BMP pathways in human muscle cells","authors":"Chloé Richard , Charlène Pourpe , Guillaume Fourneaux , Gwendal Cueff , Laurent Parry , Cécile Coudy-Gandilhon , Jonas Kindberg , Alina L. Evans , Andrea Miller , Guillemette Gauquelin-Koch , Christophe Tatout , Cécile Polge , Daniel Taillandier , Fabrice Bertile , Etienne Lefai , Lydie Combaret","doi":"10.1016/j.biocel.2025.106864","DOIUrl":"10.1016/j.biocel.2025.106864","url":null,"abstract":"<div><div>Muscle atrophy is observed in several pathophysiological situations, including physical inactivity, leading to negative health consequences, without any effective treatment currently available. Conversely, brown bears resist muscle atrophy during hibernation, despite prolonged physical inactivity and fasting. We previously reported that hibernating brown bear serum increases protein content in human myotubes and inhibits proteolysis. To go further, we deciphered here the transcriptional effects of brown bear serum in human myotubes using large-scale transcriptomics. After 48 h, the winter-hibernating bear serum (WBS) induced a specific transcriptomic program, affecting mostly biological pathways related to muscle growth and BMP signalling, compared to the summer-active bear (SBS) serum. WBS predominantly reduced, at mRNA and protein levels, activators and inhibitors of BMP signalling, which is associated with muscle mass maintenance. Moreover, BMP activity was more responsive to a stimulation by BMP7 at supra-physiological concentrations in human myotubes cultured in WBS versus SBS conditions. Meanwhile, WBS also up-regulated expression of genes encoding repressors of the pro-atrophic TGF-β pathway, decreased phosphorylated SMAD3 nuclear protein levels, and down-regulated TGF-β target genes. Furthermore, WBS treatment resulted in reduced TGF-β signalling responsiveness in human myotubes stimulated with TGF-β3 at physiological concentrations. Overall, even though WBS induced larger transcriptomic changes in the BMP compared to TGF-β pathway, the functional consequences were more pronounced for the TGF-β pathway with a marked inhibition. This study suggests that bioactive compounds in WBS may protect human muscle cells during catabolic situations, by regulating the TGF-β/BMP balance. These findings open new perspectives for therapies targeting muscle atrophy.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106864"},"PeriodicalIF":2.8,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145096627","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-16DOI: 10.1016/j.biocel.2025.106863
Wei Huang , Guanhua Deng , Qinghua Zhang, Fengquan Lv, Dehuan Xie, Chen Ren, Shasha Du, Peixin Tan
<div><h3>Background</h3><div>Radiation induced lung injury (RILI) is a common complication in patients undergoing thoracic radiotherapy. At present, there are no effective early diagnostic biomarkers, and clinical treatment methods are very limited, which poses a huge challenge to the management of cancer patients. Oxidative stress has been recognized as a key mediator of aging and disease. Therefore, this study integrated multiple omics data in mice and advanced bioinformatics and machine learning methods to systematically analyze the molecular features associated with oxidative stress, and screened for clinically relevant biomarkers and molecular mechanisms of RILI. This study aims to provide a timely and practical theoretical basis for the early diagnosis and targeted intervention of RILI.</div></div><div><h3>Method</h3><div>We implemented a comprehensive approach that integrated both bulk RNA and single-cell RNA sequencing analyses, utilizing advanced bioinformatics methodologies. These encompassed techniques aimed at eliminating batch effects to facilitate smooth data integration, executing differential expression analyses, and applying weighted gene co-expression network analysis (WGCNA). Furthermore, we developed a diagnostic model for RILI utilizing random forest and support vector machine (SVM) algorithms. We also conducted Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA). To evaluate immune cell infiltration, we employed Single-Sample Gene-Set Enrichment Analysis (ssGSEA) alongside the CIBERSORT algorithm. We then investigated the expression and interactions of module genes across various cell populations utilizing data derived from single-cell RNA sequencing. Ultimately, the expression of module genes in irradiated lung tissues were validate by reverse transcription–polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC).</div></div><div><h3>Results</h3><div>Our study identified a total of 286 differentially expressed genes (DEGs). Among these, we confirmed 61 genes related to oxidative stress (OSRDEGs). We constructed nine co-expression modules, four of which showed a significant association with RILI, encompassing 53 genes from these modules. A diagnostic model with AUC over 0.9 was constructed and further refined to include five key genes: Stk4, Aaas, Ets1, Sesn2, and Kit, which were validated for accuracy through LASSO regression. The model genes were found to be enriched in crucial pathways, particularly the MAPK signaling pathway. A direct relationship between Ets1 and Kit was found, which extended to 20 functionally similar proteins identified through GeneMANIA. Additionally, we noted significant changes in the infiltration patterns of 13 immune cell types, including Activated B cells and Activated CD4 T cells. Sens2 and Kit were found highly expressed in granulocytes and endothelial cells, respectively. In mouse models of RILI, Sesn2 and Aaas were significantl
{"title":"Integrated bulk and single-cell RNA sequencing identifies oxidative stress signatures of radiation-induced lung injury in mice through machine learning","authors":"Wei Huang , Guanhua Deng , Qinghua Zhang, Fengquan Lv, Dehuan Xie, Chen Ren, Shasha Du, Peixin Tan","doi":"10.1016/j.biocel.2025.106863","DOIUrl":"10.1016/j.biocel.2025.106863","url":null,"abstract":"<div><h3>Background</h3><div>Radiation induced lung injury (RILI) is a common complication in patients undergoing thoracic radiotherapy. At present, there are no effective early diagnostic biomarkers, and clinical treatment methods are very limited, which poses a huge challenge to the management of cancer patients. Oxidative stress has been recognized as a key mediator of aging and disease. Therefore, this study integrated multiple omics data in mice and advanced bioinformatics and machine learning methods to systematically analyze the molecular features associated with oxidative stress, and screened for clinically relevant biomarkers and molecular mechanisms of RILI. This study aims to provide a timely and practical theoretical basis for the early diagnosis and targeted intervention of RILI.</div></div><div><h3>Method</h3><div>We implemented a comprehensive approach that integrated both bulk RNA and single-cell RNA sequencing analyses, utilizing advanced bioinformatics methodologies. These encompassed techniques aimed at eliminating batch effects to facilitate smooth data integration, executing differential expression analyses, and applying weighted gene co-expression network analysis (WGCNA). Furthermore, we developed a diagnostic model for RILI utilizing random forest and support vector machine (SVM) algorithms. We also conducted Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and Gene Set Enrichment Analysis (GSEA). To evaluate immune cell infiltration, we employed Single-Sample Gene-Set Enrichment Analysis (ssGSEA) alongside the CIBERSORT algorithm. We then investigated the expression and interactions of module genes across various cell populations utilizing data derived from single-cell RNA sequencing. Ultimately, the expression of module genes in irradiated lung tissues were validate by reverse transcription–polymerase chain reaction (RT-PCR) and immunohistochemistry (IHC).</div></div><div><h3>Results</h3><div>Our study identified a total of 286 differentially expressed genes (DEGs). Among these, we confirmed 61 genes related to oxidative stress (OSRDEGs). We constructed nine co-expression modules, four of which showed a significant association with RILI, encompassing 53 genes from these modules. A diagnostic model with AUC over 0.9 was constructed and further refined to include five key genes: Stk4, Aaas, Ets1, Sesn2, and Kit, which were validated for accuracy through LASSO regression. The model genes were found to be enriched in crucial pathways, particularly the MAPK signaling pathway. A direct relationship between Ets1 and Kit was found, which extended to 20 functionally similar proteins identified through GeneMANIA. Additionally, we noted significant changes in the infiltration patterns of 13 immune cell types, including Activated B cells and Activated CD4 T cells. Sens2 and Kit were found highly expressed in granulocytes and endothelial cells, respectively. In mouse models of RILI, Sesn2 and Aaas were significantl","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106863"},"PeriodicalIF":2.8,"publicationDate":"2025-09-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145087980","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2025-09-04DOI: 10.1016/j.biocel.2025.106861
Meiqi Sun , Hua Fang , Jiashu Zhang , Mengyao Wang , Peng Jiang , LiLi Ma , Huanyu Jin , Wei Zhang
Silicosis is a fatal occupational lung disease characterized by persistent inflammation and irreversible fibrosis. However, the pathogenesis of silicosis is currently unclear. In this study, a mouse model of silicosis was established by intranasal instillation of silica, and transcriptomic alterations in lung tissues were assessed by mRNA-sequencing. Cholesterol 25-hydroxylase (Ch25h) was upregulated in silicotic lung tissues and alveolar macrophages. Lentivirus-mediated Ch25h knockdown was then employed to assess its functional role in vivo. It was found that Ch25h knockdown alleviated associated pathological changes, including pulmonary injury and fibrosis. Additionally, Ch25h significantly modulated NLRP3 inflammasome activity in vivo and in vitro. Knockdown of Ch25h inhibited the secretion of inflammatory factor (IL-1α, IL-1β, and IL-18), decreased the protein level of cleaved caspase-1 and GSDMD-N in macrophages, and reduced potassium ion efflux and lactate dehydrogenase (LDH) release. Notably, ASC (apoptosis-related spotted protein) oligomerization was suppressed by Ch25h downregulation, suggesting that Ch25h was required for the inflammasome assembly. Our findings suggest that Ch25h may contribute to silicosis development by regulating NLRP3 inflammasome activation and pyroptosis, warranting further investigation as a possible therapeutic target.
{"title":"Resistance to silicosis progression in mice with Ch25h downregulation: The involvement of NLRP3 inflammasome","authors":"Meiqi Sun , Hua Fang , Jiashu Zhang , Mengyao Wang , Peng Jiang , LiLi Ma , Huanyu Jin , Wei Zhang","doi":"10.1016/j.biocel.2025.106861","DOIUrl":"10.1016/j.biocel.2025.106861","url":null,"abstract":"<div><div>Silicosis is a fatal occupational lung disease characterized by persistent inflammation and irreversible fibrosis. However, the pathogenesis of silicosis is currently unclear. In this study, a mouse model of silicosis was established by intranasal instillation of silica, and transcriptomic alterations in lung tissues were assessed by mRNA-sequencing. Cholesterol 25-hydroxylase (Ch25h) was upregulated in silicotic lung tissues and alveolar macrophages. Lentivirus-mediated Ch25h knockdown was then employed to assess its functional role in vivo. It was found that Ch25h knockdown alleviated associated pathological changes, including pulmonary injury and fibrosis. Additionally, Ch25h significantly modulated NLRP3 inflammasome activity in vivo and in vitro. Knockdown of Ch25h inhibited the secretion of inflammatory factor (IL-1α, IL-1β, and IL-18), decreased the protein level of cleaved caspase-1 and GSDMD-N in macrophages, and reduced potassium ion efflux and lactate dehydrogenase (LDH) release. Notably, ASC (apoptosis-related spotted protein) oligomerization was suppressed by Ch25h downregulation, suggesting that Ch25h was required for the inflammasome assembly. Our findings suggest that Ch25h may contribute to silicosis development by regulating NLRP3 inflammasome activation and pyroptosis, warranting further investigation as a possible therapeutic target.</div></div>","PeriodicalId":50335,"journal":{"name":"International Journal of Biochemistry & Cell Biology","volume":"189 ","pages":"Article 106861"},"PeriodicalIF":2.8,"publicationDate":"2025-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145005125","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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