Background Age-related cataract, which presents as a cloudy lens, is the primary cause of vision impairment worldwide and can cause more than 80% senile blindness. Previous studies mainly explored the profile of lens proteins at a low concentration because of technical limitations, which could not reflect physiological status. This study focuses on protein stability changes with ageing under physiological conditions using a novel equipment, Unchained Labs (Uncle), to evaluate protein thermal stability. Methods Samples were assessed through Unchained Labs, size-exclusion chromatography, western blot and biophysics approaches including the Thioflavin T, ultraviolet and internal fluorescence. Results With age, the melting temperature value shifted from 67.8°C in the young group to 64.2°C in the aged group. Meanwhile, crystallin may form more isomeric oligomers and easy to be degraded in aged lenses. The spectroscopic and size-exclusion chromatography results show a higher solubility after administrated with lanosterol under the environmental stress. Conclusion We are the first to explore rabbit lens protein stability changes with ageing using biophysical methods under physiological conditions, and this study can conclude that the structural stability and solubility of lens proteins decrease with ageing. Additionally, lanosterol could aid in resolving protein aggregation, making it a potential therapeutic option for cataracts. So, this study provides cataract models for anti-cataract drug developments Data are available upon reasonable request. All data relevant to the study are included in the article or uploaded as supplementary information. None.

中文翻译：

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生理条件下晶状体蛋白稳定性随衰老变化的新见解

背景 年龄相关性白内障表现为混浊晶状体，是全世界视力损害的主要原因，可导致 80% 以上的老年失明。由于技术限制，以往的研究主要探讨低浓度晶状体蛋白的概况，不能反映生理状态。本研究使用新型设备 Unchained Labs (Uncle) 关注生理条件下蛋白质稳定性随老化的变化，以评估蛋白质的热稳定性。方法 通过 Unchained Labs、尺寸排阻色谱、蛋白质印迹和生物物理学方法（包括硫黄素 T、紫外线和内部荧光）评估样品。结果随着年龄的增长，熔化温度值由年轻组的67.8℃向老年组的64.2℃移动。同时，晶状体蛋白可能会形成更多异构的低聚物，并且容易在老化的镜片中降解。光谱和尺寸排阻层析结果表明，在环境压力下使用羊毛甾醇后溶解度更高。结论 我们首次在生理条件下使用生物物理方法探索兔晶状体蛋白稳定性随衰老的变化，本研究可以得出结论，晶状体蛋白的结构稳定性和溶解度随衰老而降低。此外，羊毛甾醇可以帮助解决蛋白质聚集问题，使其成为白内障的潜在治疗选择。因此，本研究为抗白内障药物开发提供了白内障模型 数据可根据合理要求提供。所有与研究相关的数据都包含在文章中或作为补充信息上传。没有任何。