AI 中文总结
本研究利用QPM探究抗坏血酸对H₂O₂诱导氧化应激的Sahiwal牛精子结构的保护作用，发现8 mg/ml抗坏血酸可显著减轻结构降解，QPM参数或可作为无标记氧化损伤指标。
AI 中文摘要
氧化应激是辅助生殖技术（ART）中低生育结局的关键因素，也是导致精子质量差的原因。常规评估依赖明场显微镜，其缺乏定量灵敏度，无法在不使用外源性造影剂的情况下解析亚细胞结构和生物物理变化，而外源性造影剂可能引入细胞毒性并损害细胞活力。本研究采用QPM作为无标记、高通量方法，探究抗坏血酸如何减轻Sahiwal牛精子的氧化结构降解。为模拟该病理状况，使用过氧化氢（H₂O₂）诱导严重氧化应激，测试两种剂量的抗坏血酸（1 mg/ml和8 mg/ml），以评估其对结构受损精子的剂量依赖性抗氧化恢复作用。QPM被用于量化关键生物物理参数的结构变化，包括干质量、光学厚度、体积、表面积、球形度、表面积体积比以及细胞内纹理参数。在急性氧化应激下，牛精子的光学厚度和干质量显著降低，同时细胞内结构组织发生可测量变化，这些共同表明氧化应激诱导的形态降解。与抗坏血酸共处理可实现部分、剂量依赖性的结构保护，与1 mg/ml处理组相比，8 mg/ml配方对这些结构变化的衰减具有统计学意义。这些发现表明，QPM衍生的生物物理参数有望作为无标记结构指标，用于表征牛精子的氧化损伤。未来研究应纳入功能验证，以确定这种结构保护是否能改善ART的结局。
英文摘要
Oxidative stress is a key factor in low fertility outcomes during assisted reproduction technology (ART) and contributes to poor sperm quality. Conventional assessment relies on bright-field microscopy, which lacks the quantitative sensitivity to resolve subcellular structural and biophysical changes without exogenous contrast agents, a requirement that can introduce cytotoxic effects and compromise cell viability. This study uses QPM as a label-free, high-throughput method to demonstrate how ascorbic acid reduces the impact of oxidative structural degradation in Sahiwal bovine spermatozoa. To illustrate this pathology, severe oxidative stress was experimentally induced using hydrogen peroxide (H2O2), and two doses of ascorbic acid (1 mg/ml and 8 mg/ml) were tested to evaluate dose-dependent antioxidant recovery in structurally damaged sperm. QPM was used to quantify structural changes in key biophysical parameters, including dry mass, optical thickness, volume, surface area, sphericity, and surface area-to-volume ratio, as well as intracellular texture parameters. Under acute oxidative stress, bovine spermatozoa exhibited significant reductions in optical thickness and dry mass, alongside measurable changes in intracellular structural organization, collectively indicative of oxidative stress-induced morphological degradation. Co-treatment with ascorbic acid resulted in partial, dose-dependent structural preservation, with the 8 mg/ml formulation demonstrating statistically significant attenuation of these structural changes compared to the 1 mg/ml treatment group. These findings suggest that QPM-derived biophysical parameters may serve as promising, label-free structural indicators for characterizing oxidative damage in bovine spermatozoa. Future studies should incorporate functional validation to determine whether this structural preservation translates to improved outcomes in ART.
Comments19 pages, 4 figures
