SERPINA3K Plays Antioxidant Roles in Cultured Pterygial Epithelial Cells through Regulating ROS System

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.

Proteome analysis of testis from infertile protein C inhibitor‐deficient mice reveals novel changes in serpin processing and prostaglandin metabolism

“…This latter serpin was first identified as a specific inhibitor of tissue kallikrein, which was also described as an interaction partner of PCI . SerpinA3K additionally has antiinflammatory, antifibrotic, and antioxidative influences on the retina []. The same pattern of high‐ and low‐molecular‐weight protein species of serpinA3K revealed here (Fig.…”

Allicin attenuates H2O2-induced cytotoxicity in retinal pigmented epithelial cells by regulating the levels of reactive oxygen species

“…The present study detected the expression levels of NOX4, which is the key enzyme of the ROS generation system (23,24), SOD, which is an important biomarker of oxidative stress (25), and NQO1, which is an important antioxidant enzyme (26). Treatment with allicin (20 µg/ml) significantly downregulated the mRNA and protein expression levels of NOX4 (mRNA, P=0.031; protein, P=0.003) following exposure of the cells to 250 µM H 2 O 2 for 24 h (Fig.…”

“…Therefore, the present study hypothesized that allicin may exert various biological abilities in order to protect against oxidative stress in various cell lines and tissues. Furthermore, NQO1 is an important enzyme, which has an essential antioxidant role (25). The protein expression levels of NQO1 were upregulated in H 2 O 2 -stimulated ARPE-19 cells following allicin pretreatment.…”

Protective Role of Kallistatin in Vascular and Organ Injury

“…61,62 In cultured endothelial cells, kallistatin via its heparin-binding site antagonized TNF-α and TGF-β-induced NADPH oxidase activity and expression, and thus ROS formation, 22,36,38 whereas kallistatin's active site was crucial for stimulating the synthesis of the antioxidant proteins eNOS, SIRT1 and FoxO1. 22 Furthermore, kallistatin exhibited antioxidant activity in cultured pterygium epithelial cells by increasing expression of superoxide dismutase 2 (SOD2), 63 which is mediated by FoxO1. 64 Many of kallistatin's antioxidant effects occur through NO, as it can inhibit NAD(P)H oxidase activity.…”