SERPINA3K plays antioxidant roles in cultured pterygial epithelial cells through regulating ROS system

PLoS One. 2014 Oct 8;9(10):e108859. doi: 10.1371/journal.pone.0108859. eCollection 2014.

Abstract

We recently demonstrated that SERPINA3K, a serine proteinase inhibitor, has antioxidant activity in the cornea. Here we investigated the antioxidant effects of SERPINA3K on the pterygial, which is partially caused by oxidative stress in pathogenesis. The head part of primary pterygial tissue was dissected and then cultured in keratinocyte serum-free defined medium (KSFM). The cultured pterygial epithelial cells (PECs) were treated with SERPINA3K. The cell proliferation and migration of PECs were measured and analyzed. Western blot and quantitative real-time polymerase chain reaction (PCR) assay were performed. It showed that SERPINA3K significantly suppressed the cell proliferation of PECs in a concentration-dependent manner, compared with cultured human conjunctival epithelial cells. SERPINA3K also inhibited the cell migration of PECs. Towards its underlying mechanism, SERPINA3K had antioxidant activities on the PECs by significantly inhibiting NADPH oxidase 4 (NOX4), which is an important enzyme of ROS generation, and by elevating the levels of key antioxidant factors of ROS: such as NAD(P)H dehydrogenase (quinone 1) (NQO1), NF-E2-related factor-2 (NRF2) and superoxide dismutases (SOD2). Meanwhile, SERPINA3K down-regulated the key effectors of Wnt signaling pathway: β-catenin, nonphospho-β-catenin, and low-density lipoprotein receptor-related protein 6 (LRP6). We provided novel evidence that SERPINA3K had inhibitory effects on pterygium and SERPINA3K played antioxidant role via regulating the ROS system and antioxidants.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Cell Survival / drug effects
  • Cells, Cultured
  • Child
  • Child, Preschool
  • Epithelial Cells / cytology
  • Epithelial Cells / drug effects*
  • Epithelial Cells / metabolism*
  • Female
  • Humans
  • Male
  • Middle Aged
  • NAD(P)H Dehydrogenase (Quinone) / metabolism
  • NADPH Oxidase 4
  • NADPH Oxidases / metabolism
  • Pterygium / metabolism*
  • Reactive Oxygen Species / metabolism*
  • Serpins / pharmacology*
  • Superoxide Dismutase / metabolism

Substances

  • Reactive Oxygen Species
  • Serpins
  • kallistatin
  • Superoxide Dismutase
  • superoxide dismutase 2
  • NADPH Oxidase 4
  • NADPH Oxidases
  • NOX4 protein, human
  • NAD(P)H Dehydrogenase (Quinone)
  • NQO1 protein, human

Grants and funding

This project was supported by a grant from Natural Science Foundation of China (grant No. 81170818) and by a grant from National High Technology Research and Development Program of China (grant No. 2012AA020507). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.