Unveiling metabolic pathways in the hyperglycemic bone: bioenergetic and proteomic analysis of the bone tissue exposed to acute and chronic high glucose.
Rita Araújo, Raquel L Bernardino, Mariana P Monteiro, Pedro S Gomes
{"title":"Unveiling metabolic pathways in the hyperglycemic bone: bioenergetic and proteomic analysis of the bone tissue exposed to acute and chronic high glucose.","authors":"Rita Araújo, Raquel L Bernardino, Mariana P Monteiro, Pedro S Gomes","doi":"10.1186/s10020-025-01251-0","DOIUrl":null,"url":null,"abstract":"<p><strong>Background: </strong>Bone fragility due to poor glycemic control is a recognized complication of diabetes, but the mechanisms underlying diabetic bone disease remain poorly understood. Despite the importance of bioenergetics in tissue functionality, the impact of hyperglycemia on bone bioenergetics has not been previously investigated.</p><p><strong>Objective: </strong>To determine the effects of high glucose exposure on energy metabolism and structural integrity in bone tissue using an ex vivo organotypic culture model of embryonic chick femur.</p><p><strong>Methods: </strong>Femora from eleven-day-old Gallus gallus embryos were cultured for eleven days under physiological glucose conditions (5.5 mM, NG), chronic high glucose exposure (25 mM, HG-C), or acute high glucose exposure (25 mM, HG-A). Bioenergetic assessments (Seahorse assays), proteomic analysis (liquid chromatography-mass spectrometry), histomorphometric and microtomographic evaluations, and oxidative stress measurements (carbonyl content assay) were performed. Statistical analyses were conducted using IBM® SPSS® Statistics (v26.0). The Mann-Whitney nonparametric test was used for group comparisons in microtomographic analysis, ALP activity, and carbonyl content assays. For Seahorse assay results, ANOVA with Tukey's post-hoc test was applied after confirming data homoscedasticity with Levene's test.</p><p><strong>Results: </strong>Chronic high glucose exposure reduced bone mineral deposition, altered histomorphometric indices, and suppressed key osteochondral development regulators. Acute high glucose exposure enhanced glycolysis and oxidative phosphorylation, while chronic exposure caused oxygen consumption uncoupling, increased ROS generation, and downregulated mitochondrial proteins critical for bioenergetics. Elevated oxidative stress was confirmed in the chronic high glucose group.</p><p><strong>Conclusion: </strong>Chronic high glucose exposure disrupted bone bioenergetics, induced mitochondrial dysfunction, and compromised bone structural integrity, emphasizing the metabolic impact of hyperglycemia in diabetic bone disease.</p>","PeriodicalId":18813,"journal":{"name":"Molecular Medicine","volume":"31 1","pages":"194"},"PeriodicalIF":6.0000,"publicationDate":"2025-05-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12084916/pdf/","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Medicine","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1186/s10020-025-01251-0","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Background: Bone fragility due to poor glycemic control is a recognized complication of diabetes, but the mechanisms underlying diabetic bone disease remain poorly understood. Despite the importance of bioenergetics in tissue functionality, the impact of hyperglycemia on bone bioenergetics has not been previously investigated.
Objective: To determine the effects of high glucose exposure on energy metabolism and structural integrity in bone tissue using an ex vivo organotypic culture model of embryonic chick femur.
Methods: Femora from eleven-day-old Gallus gallus embryos were cultured for eleven days under physiological glucose conditions (5.5 mM, NG), chronic high glucose exposure (25 mM, HG-C), or acute high glucose exposure (25 mM, HG-A). Bioenergetic assessments (Seahorse assays), proteomic analysis (liquid chromatography-mass spectrometry), histomorphometric and microtomographic evaluations, and oxidative stress measurements (carbonyl content assay) were performed. Statistical analyses were conducted using IBM® SPSS® Statistics (v26.0). The Mann-Whitney nonparametric test was used for group comparisons in microtomographic analysis, ALP activity, and carbonyl content assays. For Seahorse assay results, ANOVA with Tukey's post-hoc test was applied after confirming data homoscedasticity with Levene's test.
Results: Chronic high glucose exposure reduced bone mineral deposition, altered histomorphometric indices, and suppressed key osteochondral development regulators. Acute high glucose exposure enhanced glycolysis and oxidative phosphorylation, while chronic exposure caused oxygen consumption uncoupling, increased ROS generation, and downregulated mitochondrial proteins critical for bioenergetics. Elevated oxidative stress was confirmed in the chronic high glucose group.
Conclusion: Chronic high glucose exposure disrupted bone bioenergetics, induced mitochondrial dysfunction, and compromised bone structural integrity, emphasizing the metabolic impact of hyperglycemia in diabetic bone disease.
背景：血糖控制不良导致的骨质疏松是公认的糖尿病并发症，但糖尿病性骨病的机制尚不清楚。尽管生物能量在组织功能中的重要性，但高血糖对骨生物能量的影响尚未被研究过。目的：建立鸡胚胎股骨离体培养模型，探讨高糖暴露对鸡胚胎股骨能量代谢和骨组织结构完整性的影响。方法：将11日龄母鸡胚胎的股骨在生理葡萄糖条件（5.5 mM， NG）、慢性高糖暴露（25 mM, HG-C）或急性高糖暴露（25 mM, HG-A）下培养11天。进行生物能量评估（海马试验）、蛋白质组学分析（液相色谱-质谱）、组织形态和显微层析评估以及氧化应激测量（羰基含量测定）。采用IBM®SPSS®Statistics （v26.0）进行统计分析。曼-惠特尼非参数检验用于显微层析分析、ALP活性和羰基含量测定的组比较。海马试验结果在用Levene检验确认数据均方差后，采用Tukey事后检验进行方差分析。结果：慢性高糖暴露减少骨矿物质沉积，改变组织形态学指标，抑制关键骨软骨发育调节因子。急性高糖暴露增强糖酵解和氧化磷酸化，而慢性暴露导致氧气消耗解偶联，ROS生成增加，并下调对生物能量学至关重要的线粒体蛋白。慢性高糖组氧化应激升高。结论：慢性高糖暴露破坏骨生物能量，诱导线粒体功能障碍，破坏骨结构完整性，强调高血糖在糖尿病骨病中的代谢影响。
期刊介绍：
Molecular Medicine is an open access journal that focuses on publishing recent findings related to disease pathogenesis at the molecular or physiological level. These insights can potentially contribute to the development of specific tools for disease diagnosis, treatment, or prevention. The journal considers manuscripts that present material pertinent to the genetic, molecular, or cellular underpinnings of critical physiological or disease processes. Submissions to Molecular Medicine are expected to elucidate the broader implications of the research findings for human disease and medicine in a manner that is accessible to a wide audience.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
