Quantitative proteomics analysis with iTRAQ in human lenses with nuclear cataracts of different axial lengths.

摘要: PURPOSE The goal of this study was to identify and quantify the differentially expressed proteins in human nuclear cataract with different axial lengths. METHODS Thirty-six samples lens nuclei hardness grade III or IV were obtained during surgery extracapsular extraction (ECCE). Six healthy transparent from fresh cadaver eyes corneal transplantation surgery. divided into seven groups (six lenses each group) according optic axis: Group A (mean length 28.7±1.5 mm; average age 59.8±1.9 years), B 23.0±0.4 60.3±2.5 C 19.9±0.5 55.1±2.5 D 28.7±1.4 58.0±4.0 E 23.0±0.3 56.9±4.2 F 20.7±0.6 57.6±5.3 years). six included a younger group standard axes, G 23.0±0.5 34.7±4.2 years).Water-soluble, water-insoluble, water-insoluble-urea-soluble protein fractions extracted samples. three-part individual combined form total sample. proteomic profiles analyzed using 8-plex isobaric tagging for relative absolute quantification (iTRAQ) labeling two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS). data ProteinPilot software peptide matching, identification, quantification. Differentially validated western blotting. RESULTS We employed biological technical replicates selected intersection two sets results, which 40 proteins. From identified, as closely related length. gap junction alpha-3 protein, beta-crystallin B2, T-complex 1 subunit beta, gamma-enolase, pyruvate kinase isozymes M1/M2, sorbitol dehydrogenase. Levels B2 expression decreased cataracts longer results spectrometric analysis consistent blot validation. CONCLUSION discovery these provides valuable clues understanding pathogenesis axial-related cataract. indicate that (CRBB2) may be involved pathogenesis. Further studies are needed investigate correlation between CRBB2