Mitra Ronodeep, Pentland Kaleigh, Kolev Svilen, Eden Matthew, Levine Erel, Oakes Jessica M, Ebong Eno E
Department of Chemical Engineering, Northeastern University, Boston, MA, United States.
Department of Bioengineering, Northeastern University, Boston, MA, United States.
Life Sci. 2025 Sep 15;377:123662. doi: 10.1016/j.lfs.2025.123662. Epub 2025 Apr 23.
Endothelial cell (EC) glycocalyx (GCX) shedding from disturbed blood flow and chemical factors leads to low-density lipoprotein infiltration, reduced nitric oxide synthesis, vascular dysfunction and atherosclerosis. This study evaluates a therapy combining sphingosine-1-phosphate (S1P) and heparin (heparan sulfate derivative). We hypothesized that heparin/S1P co-treatment repairs mechanically damaged EC GCX in disturbed flow (DF) regions and restores anti-atherosclerotic mechanotransduction to treat cardiovascular disease.
We used a parallel-plate flow chamber to simulate flow conditions in vitro and a partial carotid ligation mouse model to mimic DF in vivo. Heparin and albumin-bound S1P were administered to assess their reparative effects on the endothelial GCX. Fluorescent staining, confocal microscopy, and ultrasound evaluated endothelial cell function and endothelial-dependent vascular function. Barrier functionality was assessed via macrophage uptake. Heparin/S1P mechanism-of-action insights were gained through fluid dynamics simulations and staining of GCX synthesis enzyme and S1P receptor. Statistical analyses validated the results.
The in vitro data showed that heparin/S1P therapy improves DF-conditioned ECs by restoring GCX and elevating vasodilator eNOS (endothelial-type nitric oxide synthase) expression. In vivo studies confirmed GCX degradation, vessel inflammation, hyperpermeability, and wall thickening in the mouse model's partially ligated left carotid artery. Heparin/S1P treatment restored GCX thickness and coverage, reduced inflammation and hyperpermeability, and inhibited vessel wall thickening.
This work introduces a new approach to regenerating the EC GCX and restoring its function in ECs under DF conditions, offering a groundbreaking solution for preventing cardiovascular diseases like atherosclerosis.
血流紊乱和化学因素导致内皮细胞（EC）糖萼（GCX）脱落，进而引起低密度脂蛋白浸润、一氧化氮合成减少、血管功能障碍和动脉粥样硬化。本研究评估了鞘氨醇-1-磷酸（S1P）和肝素（硫酸乙酰肝素衍生物）联合治疗方法。我们假设肝素/S1P联合治疗可修复血流紊乱（DF）区域机械损伤的EC GCX，并恢复抗动脉粥样硬化的机械转导以治疗心血管疾病。
我们使用平行板流动腔在体外模拟血流条件，并使用部分颈动脉结扎小鼠模型在体内模拟DF。给予肝素和白蛋白结合的S1P，以评估它们对内皮GCX的修复作用。通过荧光染色、共聚焦显微镜和超声评估内皮细胞功能和内皮依赖性血管功能。通过巨噬细胞摄取评估屏障功能。通过流体动力学模拟以及GCX合成酶和S1P受体染色深入了解肝素/S1P的作用机制。统计分析验证了结果。
体外数据表明，肝素/S1P治疗可通过恢复GCX和提高血管舒张剂内皮型一氧化氮合酶（eNOS）表达来改善DF条件下的EC。体内研究证实了小鼠模型部分结扎的左颈动脉中GCX降解、血管炎症、高通透性和管壁增厚。肝素/S1P治疗可恢复GCX厚度和覆盖范围，减轻炎症和高通透性，并抑制血管壁增厚。
这项工作引入了一种在DF条件下再生EC GCX并恢复其在EC中功能的新方法，为预防动脉粥样硬化等心血管疾病提供了开创性的解决方案。
