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
- lens and zonules
- experimental – animal models
Data availability statement
Data are available upon reasonable request. All data relevant to the study are included in the article or uploaded as supplementary information. None.
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CL, JX and CF are joint first authors.
CL, JX and CF contributed equally.
Contributors CL acquired and analysed the data and drafted the manuscript. CL, JX and CF performed the experiments and revised the manuscript. KY and XC conceived and designed the study, interpreted and analysed the data, and revised the manuscript. All the authors have read and approved the final manuscript.
Funding This study was supported by the National Natural Science Foundation of China (number 31872724, number 81700815 and number 81900837) and Zhejiang Natural Science Funds for Distinguished Young Scholar (number LR21H120001).
Competing interests None declared.
Provenance and peer review Not commissioned; externally peer reviewed.
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