Abstract
Background: Corneal neovascularization (angiogenesis and lymphangiogenesis) compromises corneal transparency and transplant survival. The contribution and molecular mechanisms of three kinds of corneal host cells, including corneal epithelial, stromal, and endothelial cells, involved in corneal neovascularization are currently unclear. Methods: Liquid chromatography-mass spectrometry, immunoblotting, and ELISA were used to screen and identify potential neovascularization related factors in human full thickness vascularized corneal tissues. In vitro inflammation model, plasmid transfection, recombinant stimulation, tube formation assay, spheroid sprouting assay, and in vivo corneal neovascularization model were employed to clarify the contribution of three corneal host cells to neovascularization and identify the key proteins involved. Findings: All three kinds of corneal host cells could influence corneal neovascularization, but varied in degrees. Matrix metalloproteinase-9 (MMP-9) in human corneal epithelial cells, MMP-2, and alpha-crystallin A chain (CRYAA) in human corneal stromal cells, and MMP-2 and galectin-8 in human corneal endothelial cells might be key proteins participating into corneal neovascularization, respectively. Interpretation: These data highlight that both the effects of key proteins and cornea host cells involved should be considered for therapy of corneal neovascularization. Funding Statement: This study was supported by the Nature Science Foundation of China (81470618), the Scientific Research Foundation of First Affiliated Hospital of Harbin Medical University (2017B013), the Major Program of Applied Technology Research and Development Plan of Heilongjiang Province (GY2016ZB0159), the Scientific Research Fund of Heilongjiang Provincial Education Department (12521262), the Scientific Research Fund of Heilongjiang Provincial Health Bureau (2011-031), the Special Fund for the Doctoral Program of Higher Education (20132307120035), the Natural Science Foundation of China (81300728), the Natural Science Foundation of Heilongjiang Province of China (QC2010113), and the Postdoctoral Grant of Heilongjiang Province (LBH-Q12038). Declaration of Interests: The authors have no conflicts of interest to declare. Ethics Approval Statement: All experiments were conducted in accordance with Declaration of Helsinki Principles, and all experiments were performed with the approval of the Internal Review Board of Harbin Medical University. We obtained the informed consents from all patients prior to mortality or their family members following mortality for inclusion of autopsy specimens. The rats were housed in the Harbin Medical University Experimental Animal Resources Center in accordance with National Institutes of Health guidelines and operated upon using a research protocol approved by the Institutional Animal Care and Use Committee.
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