Jiang Zhao, Alamuri Tharun T, Yang Darren L, Annarino Tyler, Muir Eric R, Duong Tim Q
Department of Radiology, Stony Brook Medicine, Stony Brook, NY, USA.
Department of Radiology, University of North Carolina, Chapel Hill, NC, USA.
Brain Res. 2025 Nov 1;1866:149940. doi: 10.1016/j.brainres.2025.149940. Epub 2025 Sep 9.
Ischemic stroke remains a leading cause of death and disability worldwide, with limited effective treatments due to the complexity of its pathophysiology. Molecular hydrogen (H) and minocycline (M), both possessing anti-inflammatory and antioxidant properties, have shown individual neuroprotective potential in preclinical models. However, the optimal therapeutic dosing of H, particularly in combination with other agents, remains undefined.
This study aimed to (1) determine the dose-response relationship of hydrogen-enriched water in a rat model of transient middle cerebral artery occlusion (MCAO), and (2) evaluate whether optimized H dosing combined with minocycline provides superior neuroprotection compared to H monotherapy.
Sixty-six male and female Sprague-Dawley rats underwent 60-minute MCAO followed by treatment with varying doses (5-30 mL/kg) of hydrogen-enriched water (3.2 ppm), alone or in combination with minocycline (20 mg/kg). Treatments were administered post-reperfusion as well as on days 1 and 2. Behavioral outcomes (Garcia score) and infarct volumes (TTC staining) were assessed at 7 days post-stroke.
The optimal H dose was 20 mL/kg, which produced the highest Garcia scores and lowest infarct volumes. A dose-dependent effect was observed with a quadratic fit (R = 0.751 for Garcia scores; R = 0.289 for lesion volume). Combination therapy with H and minocycline significantly outperformed H monotherapy in both neurological recovery and infarct reduction, with no sex differences observed.
Hydrogen-enriched water shows a dose-dependent neuroprotective effect in experimental ischemic stroke, with 20 mL/kg identified as the optimal dose. Combined therapy with minocycline further enhances outcomes, supporting the potential of dual-agent strategies for improved stroke treatment. These findings provide a foundation for translational development of H-based combination therapies in clinical settings.
缺血性中风仍然是全球死亡和残疾的主要原因，由于其病理生理学的复杂性，有效治疗方法有限。分子氢（H）和米诺环素（M）都具有抗炎和抗氧化特性，在临床前模型中已显示出各自的神经保护潜力。然而，H的最佳治疗剂量，尤其是与其他药物联合使用时，仍未明确。
本研究旨在（1）确定富氢水在大鼠短暂性大脑中动脉闭塞（MCAO）模型中的剂量反应关系，以及（2）评估优化的H剂量与米诺环素联合使用是否比H单一疗法提供更好的神经保护作用。
66只雄性和雌性Sprague-Dawley大鼠接受60分钟的MCAO，然后分别用不同剂量（5-30 mL/kg）的富氢水（3.2 ppm）单独或与米诺环素（20 mg/kg）联合治疗。在再灌注后以及第1天和第2天进行治疗。在中风后7天评估行为结果（加西亚评分）和梗死体积（TTC染色）。
最佳H剂量为20 mL/kg，此时加西亚评分最高，梗死体积最小。观察到剂量依赖性效应，呈二次拟合（加西亚评分为R = 0.751；病变体积为R = 0.289）。H与米诺环素联合治疗在神经功能恢复和梗死减少方面均明显优于H单一疗法，未观察到性别差异。
富氢水在实验性缺血性中风中显示出剂量依赖性神经保护作用，确定最佳剂量为20 mL/kg。与米诺环素联合治疗可进一步改善结果，支持双药策略改善中风治疗的潜力。这些发现为基于H的联合疗法在临床环境中的转化发展提供了基础。
