Quantitative proteomics analysis by iTRAQ in human nuclear cataracts of different ages and normal lens nuclei

摘要: Purpose The goal of this study was to quantitatively identify the differentially expressed proteins in nuclear cataracts different ages and normal lens nuclei humans. Experimental design Forty-eight human nucleus samples with hardness grades III, IV were obtained during cataract surgery by extracapsular extraction. Seven transparent from fresh cadaver eyes corneal transplantation surgery. Lens divided into seven groups according age optic axis: Group A (average 80.8 ± 1.2 years), B 57.0 4.0 years), C average 80.3 4.5 years), Group D 56.9 4.2 years), E 78.1 2.5 years), F 57.6 3.3 years) G (seven lenses eyes, 34.7 4.2 years). Water-soluble, water-insoluble, water-insoluble-urea-soluble protein fractions extracted samples. The three-part individual combined form total each sample. proteomic profiles group further analyzed using 8-plex iTRAQ labeling 2D-LC-MS/MS. data ProteinPilot software for peptide matching, identification, quantification. Differentially validated Western blotting. Results We employed biological technical replicates selected intersection two results, which included 80 proteins. Nine among identified techniques. In age-related (ARNC), expression levels fatty acid-binding pterin-4-alpha-carbinolamine dehydratase upregulated, whereas alpha-crystallin chain (CRYAB), GSH synthetase, phakinin, gamma-crystallin C, phosphoglycerate kinase 1, betaine-homocysteine S-methyltransferase 1 (BHMT1), spectrin beta downregulated. These may be associated abnormal aggregation oxidative stress. synthetase CRYAB decreased age. mass spectrometric analysis results consistent blot validation. Conclusion clinical relevance The indicate that involved ARNC pathogenesis. 2D-LC-MS/MS provides new methods future studies pathological mechanisms protective drug development ARNC.