title: Oxidative stress and premature senescence in corneal endothelium following penetrating keratoplasty in an animal model.
abstract: BACKGROUND
  The purpose of this study was to address the question of how the premature senescence process may affect corneal endothelium after penetrating keratoplasty, because the quality of donor corneal endothelial cells is important for corneal transplant success.
  
  
  METHODS
  The cell senescence and induced oxidative stress in corneal endothelium were assessed using a normal-risk orthotopic mice corneal transplantation model. Senescence associated beta-galactosidase (SA-beta-Gal) staining was used to evaluate premature senescence in the endothelium of corneal allografts. Oxidative Stress and Antioxidant Defense RT(2)-PCR Arrays and in vitro experimental model using H2O2 treatment were used to investigate the possible mechanism.
  
  
  RESULTS
  SA-beta-Gal positivity was observed obviously in mice corneal endothelium of allogenic group and the levels of p16(INK4a) message and protein increased in endothelium of allogenic group compared to syngenic group. By PCR array, an oxidant-antioxidant imbalance was found in the endothelium of corneal allograft after PKP. The results from mice model were validated using human endothelium samples of corneal allograft after PKP. We also developed an in vitro experimental model using H2O2 treatment to simulate a state of oxidative stress in cultured human corneal endothelial cells (HCECs) and found that elevated ROS levels, the up-regulation of CDK inhibitors and ROS-mediated p16(INK4A) up-regulation in HCECs occur via the ASK1-p38 MAPK pathway.
  
  
  CONCLUSIONS
  Our results demonstrate the presence of oxidative stress and premature senescence in the endothelium of corneal allografts following PKP.
authors.affiliation: State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of medical Sciences, No. 5 Yanerdao Rd, Qingdao, 266071, China.
authors.forename: Xiaowen
authors.initials: X
authors.lastname: Zhao
authors.affiliation: State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of medical Sciences, No. 5 Yanerdao Rd, Qingdao, 266071, China. wye112002@126.com.|Current affiliation: Central Laboratory of the Second Affiliated Hospital, Medical College of Qingdao University, Qingdao, 266042, China. wye112002@126.com.
authors.forename: Ye
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of medical Sciences, No. 5 Yanerdao Rd, Qingdao, 266071, China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of medical Sciences, No. 5 Yanerdao Rd, Qingdao, 266071, China.
authors.forename: Suxia
authors.initials: S
authors.lastname: Li
authors.affiliation: State Key Laboratory Cultivation Base, Shandong Provincial Key Laboratory of Ophthalmology, Shandong Eye Institute, Shandong Academy of medical Sciences, No. 5 Yanerdao Rd, Qingdao, 266071, China.
authors.forename: Peng
authors.initials: P
authors.lastname: Chen
mesh_terms: D000328:Adult; D064591:Allografts; D000818:Animals; D002478:Cells, Cultured; D016922:Cellular Senescence; D019941:Cyclin-Dependent Kinase Inhibitor p16 / Q000235:genetics / Q000378:metabolism; D050760:Cyclin-Dependent Kinase Inhibitor p27 / Q000235:genetics / Q000378:metabolism; D004195:Disease Models, Animal; D004728:Endothelium, Corneal / Q000187:drug effects / Q000378:metabolism / Q000473:pathology*; D005786:Gene Expression Regulation; D006084:Graft Rejection / Q000209:etiology* / Q000235:genetics / Q000473:pathology; D006801:Humans; D006861:Hydrogen Peroxide / Q000633:toxicity; D015948:Keratoplasty, Penetrating / Q000009:adverse effects*; D048848:MAP Kinase Kinase Kinase 5 / Q000378:metabolism; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D008875:Middle Aged; D018384:Oxidative Stress; D017382:Reactive Oxygen Species / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015854:Up-Regulation; D055815:Young Adult; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D019941:Cyclin-Dependent Kinase Inhibitor p16; D017382:Reactive Oxygen Species; D050760:Cyclin-Dependent Kinase Inhibitor p27; D006861:Hydrogen Peroxide; D048051:p38 Mitogen-Activated Protein Kinases; D048848:MAP Kinase Kinase Kinase 5; C482859:Map3k5 protein, mouse; D001616:beta-Galactosidase
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