title: iTRAQ-Based Quantitative Proteomic Analysis of Tear Fluid in a Rat Penetrating Keratoplasty Model With Acute Corneal Allograft Rejection.
abstract: OBJECTIVE
  This study aimed to develop a greater understanding of the mechanisms underlying acute corneal allograft rejection by identifying differentially expressed tear proteins at defined stages and discovering potentially important proteins involved in the process.
  
  
  METHODS
  The isobaric tags for relative and absolute quantitation (iTRAQ)-two-dimensional liquid chromatography-tandem mass spectrometry (2DLC-MS/MS) technique was used to identify tear proteins showing significant alterations in a rat penetrating keratoplasty model at different time points. Bioinformatics technology was applied to analyze the significant proteins, and a potential protein was verified by Western blotting.
  
  
  RESULTS
  A total of 269 proteins were quantified, and 118 proteins were considered to be significantly altered by at least 2.0- or 0.5-fold. For gene ontology annotations, the top enrichments were neurological disease, free radical scavenging, cell death and survival, and cell movement. For pathway analyses, the top enrichments were LXR/RXR activation, acute phase response signaling, clathrin-mediated endocytosis signaling, and coagulation system. Coronin-1A was verified as a potential protein involved in the early stage of acute corneal allograft rejection.
  
  
  CONCLUSIONS
  This study first demonstrates that tear proteomics is a powerful tool for better understanding of the mechanisms underlying acute corneal rejection, and that coronin-1A in tears might be closely related to allograft rejection.
authors.affiliation: Key Laboratory of Myopia Ministry of Health, Department of Ophthalmology, Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China.
authors.forename: Feifei
authors.initials: F
authors.lastname: Huang
authors.affiliation: Key Laboratory of Myopia Ministry of Health, Department of Ophthalmology, Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China.
authors.forename: Jianjiang
authors.initials: J
authors.lastname: Xu
authors.affiliation: Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
authors.forename: Hong
authors.initials: H
authors.lastname: Jin
authors.affiliation: Key Laboratory of Myopia Ministry of Health, Department of Ophthalmology, Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China.
authors.forename: Jianwen
authors.initials: J
authors.lastname: Tan
authors.affiliation: Key Laboratory of Myopia Ministry of Health, Department of Ophthalmology, Eye, Ear, Nose and Throat Hospital, Fudan University, Shanghai, China.
authors.forename: Chaoran
authors.initials: C
authors.lastname: Zhang
mesh_terms: D000208:Acute Disease; D064591:Allografts; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002853:Chromatography, Liquid / Q000379:methods; D003316:Corneal Diseases / Q000601:surgery*; D004195:Disease Models, Animal; D005136:Eye Proteins / Q000378:metabolism*; D006084:Graft Rejection / Q000378:metabolism*; D015948:Keratoplasty, Penetrating; D008297:Male; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D053719:Tandem Mass Spectrometry / Q000379:methods; D013666:Tears / Q000378:metabolism*
chemical_list: D015415:Biomarkers; D005136:Eye Proteins; C042434:tear proteins