title: Quantitative proteomics analysis with iTRAQ in human lenses with nuclear cataracts of different axial lengths.
abstract: The goal of this study was to identify and quantify the differentially expressed proteins in human nuclear cataract with different axial lengths.
  
  
  
  Thirty-six samples of human lens nuclei with hardness grade III or IV were obtained during cataract surgery with extracapsular cataract extraction (ECCE). Six healthy transparent human lens nuclei were obtained from fresh healthy cadaver eyes during corneal transplantation surgery. The lens nuclei were divided into seven groups (six lenses in each group) according to the optic axis: Group A (mean axial length 28.7±1.5 mm; average age 59.8±1.9 years), Group B (mean axial length 23.0±0.4 mm; average age 60.3±2.5 years), Group C (mean axial length 19.9±0.5 mm; average age 55.1±2.5 years), Group D (mean axial length 28.7±1.4 mm; average age 58.0±4.0 years), Group E (mean axial length 23.0±0.3 mm; average age 56.9±4.2 years), and Group F (mean axial length 20.7±0.6 mm; average age 57.6±5.3 years). The six healthy transparent human lenses were included in a younger group with standard optic axes, Group G (mean axial length 23.0±0.5 mm; average age 34.7±4.2 years).Water-soluble, water-insoluble, and water-insoluble-urea-soluble protein fractions were extracted from the samples. The three-part protein fractions from the individual lenses were combined to form the total proteins of each sample. The proteomic profiles of each group were analyzed using 8-plex isobaric tagging for relative and absolute protein quantification (iTRAQ) labeling combined with two-dimensional liquid chromatography tandem mass spectrometry (2D-LC-MS/MS). The data were analyzed with ProteinPilot software for peptide matching, protein identification, and quantification. Differentially expressed proteins were validated with western blotting.
  
  
  
  We employed biological and technical replicates and selected the intersection of the two sets of results, which included 40 proteins. From the 40 proteins identified, six were selected as differentially expressed proteins closely related to axial length. The six proteins were gap junction alpha-3 protein, beta-crystallin B2, T-complex protein 1 subunit beta, gamma-enolase, pyruvate kinase isozymes M1/M2, and sorbitol dehydrogenase. Levels of beta-crystallin B2 expression were decreased in nuclear cataracts with longer axial length. The results of the mass spectrometric analysis were consistent with the western blot validation.
  
  
  
  The discovery of these differentially expressed proteins provides valuable clues for understanding the pathogenesis of axial-related nuclear cataract. The results indicate that beta-crystallin B2 (CRBB2) may be involved in axial-related nuclear cataract pathogenesis. Further studies are needed to investigate the correlation between CRBB2 and axial-related nuclear cataract.
authors: Department of Ophthalmology, Tangdu Hospital, Fourth Military Medical University, Xi'an, China,; Department of Ophthalmology, Shaanxi Provincial People's Hospital, Xi'an, China.
authors: Haiyan
authors: H
authors: Zhou
authors: Department of Ophthalmology, Tangdu Hospital, Fourth Military Medical University, Xi'an, China,; Chongqing Key Laboratory of Ophthalmology, Chongqing Eye Institute, the First Affiliated Hospital, Chongqing Medical University, Chongqing, China.
authors: Hong
authors: H
authors: Yan
authors: Department of Clinical Medicine, Xi'an Medical University, Xi'an, China.
authors: Weijia
authors: W
authors: Yan
authors: Department of Neurosurgery, Tangdu Hospital, Fourth Military Medical University, Xi'an, China.
authors: Xinchuan
authors: X
authors: Wang
authors: Department of Ophthalmology, Shaanxi Provincial People's Hospital, Xi'an, China.
authors: Yong
authors: Y
authors: Ma
authors: Department of Ophthalmology, Shaanxi Provincial People's Hospital, Xi'an, China.
authors: Jianping
authors: J
authors: Wang
mesh_terms: D000328:Adult; D058452:Axial Length, Eye / Q000473:pathology*; D015153:Blotting, Western; D002386:Cataract / Q000151:congenital* / Q000378:metabolism; D002853:Chromatography, Liquid; D003459:Crystallins / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005260:Female; D006801:Humans; D007907:Lens Nucleus, Crystalline / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D040901:Proteomics; D053719:Tandem Mass Spectrometry
chemical_list: D003459:Crystallins