Hydrogen rich saline treatment modulates proteomic profiles to mitigate cataract development in an N methyl N nitrosourea induced rat model

Purpose: Cataracts are associated with oxidative stress-induced damage to lens proteins. This study aims to identify Differentially Expressed Proteins (DEPs) associated with the protective effects of Hydrogen-Rich Saline (HRS) against N-Methyl-N-Nitrosourea (MNU)-induced cataracts, utilizing the antioxidant properties of hydrogen.

Methods: Sprague-Dawley rats were assigned to control, MNU-only, MNU+NS, MNU+pirenoxine, and MNU+HRS groups. Cataracts were induced with MNU (postnatal day 15), and treatments (postnatal days 8–21) included intraperitoneal injections and eye drops. Cataract severity was assessed using slit-lamp examinations, Pentacam analysis, and spectrophotometry. Proteomic analysis of lens tissues from the MNU+HRS and MNU+NS groups employed TMT labeling and mass spectrometry. DEPs were identified, grouped based on fold changes, and analyzed for GO, KEGG, and domain enrichment. Parallel Reaction Monitoring (PRM) validated selected DEPs.

Results: HRS reduced MNU-induced cataract incidence to 50% versus 100% in MNU-only and NS groups and preserved lens clarity comparable to normal controls. Proteomic analysis identified 90 upregulated and 303 downregulated proteins in the HRS-treated group versus the NS-treated group. DEPs were enriched in GO terms related to ion transport, homeostasis, and ATP hydrolysis, as well as KEGG pathways like oxidative phosphorylation and arginine biosynthesis. Domain enrichment showed links to ATPase activity and energy metabolism. DEPs were grouped into Q1–Q4, with Q1 showing enrichment in oxidative phosphorylation and metabolic pathways. PRM confirmed the downregulation of 14 stress-response and metabolic proteins in the HRS-treated group.

Conclusion: HRS mitigates MNU-induced cataracts possibly by reducing oxidative stress and downregulating stress-response and metabolic proteins.

Clinical trial number: Not applicable.

Keywords: Cataract, N-Methyl-N-Nitrosourea, Hydrogen, Oxidative Stress, Tandem Mass Spectrometry, Proteomics.

Doctor of Ophthalmology, Associate Chief Physician, Associate Professor. Member of the Ophthalmology Professional Committee of the Beijing Society of Integrated Traditional Chinese and Western Medicine, member of the Beijing Ophthalmological Society, expert member of the Popular Science Work Committee for Myopia Prevention and Control, and advisor of the Asia-Pacific Section of the International Corneal Reshape Society, as well as a member of the International Precision Refraction Association. Professional Achievements: With 20 years of experience in ophthalmic clinical work, she specializes in various complex and difficult cataract surgeries, personalized refractive cataract surgeries, LRI astigmatism correction surgery, small incision phacoemulsification combined with intraocular lens implantation, ICL implantation surgery, anti-VEGF drug treatment for age-related macular degeneration and diabetic retinopathy, and 4L therapy for childhood myopia prevention and control. She has published 6 SCI papers, 5 papers in core journals, edited 1 monograph, obtained 2 utility model invention patents, participated in 1 national natural science foundation general project, and hosted 1 provincial natural science foundation general project.