Stanniocalcin-1 protects bovine intestinal epithelial cells from oxidative stress-induced damage

J Vet Sci. 2014 Dec;15(4):475-83. doi: 10.4142/jvs.2014.15.4.475. Epub 2014 Jun 20.

Abstract

Chronic enteritis can produce an excess of reactive oxygen species resulting in cellular damage. Stanniocalcin-1(STC-1) reportedly possesses anti-oxidative activity, the aim of this study was to define more clearly the direct contribution of STC-1 to anti-oxidative stress in cattle. In this study, primary intestinal epithelial cells (IECs) were exposed to hydrogen peroxide (H2O2) for different time intervals to mimic chronic enteritis-induced cellular damage. Prior to treatment with 200 µM H2O2, the cells were transfected with a recombinant plasmid for 48 h to over-express STC-1. Acridine orange/ ethidium bromide (AO/EB) double staining and trypan blue exclusion assays were then performed to measure cell viability and apoptosis of the cells, respectively. The expression of STC-1 and apoptosis-related proteins in the cells was monitored by real-time PCR and Western blotting. The results indicated that both STC-1 mRNA and protein expression levels positively correlated with the duration of H2O2 treatment. H2O2 damaged the bovine IECs in a time-dependent manner, and this effect was attenuated by STC-1 over-expression. Furthermore, over- expression of STC-1 up-regulated Bcl-2 protein expression and slightly down-regulated caspase-3 production in the damaged cells. Findings from this study suggested that STC-1 plays a protective role in intestinal cells through an antioxidant mechanism.

Keywords: Bcl-2; chronic enteritis; oxidative damage; stanniocalcin-1.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Animals, Newborn
  • Blotting, Western / veterinary
  • Caspase 3 / genetics*
  • Caspase 3 / metabolism
  • Cattle
  • Cattle Diseases / etiology
  • Cattle Diseases / genetics*
  • Cattle Diseases / metabolism
  • Duodenum / metabolism
  • Enteritis / etiology
  • Enteritis / genetics
  • Enteritis / metabolism
  • Enteritis / veterinary*
  • Epithelial Cells / metabolism
  • Gene Expression Regulation*
  • Glycoproteins / genetics*
  • Glycoproteins / metabolism
  • Hydrogen Peroxide / pharmacology
  • Male
  • Proto-Oncogene Proteins c-bcl-2 / genetics*
  • Proto-Oncogene Proteins c-bcl-2 / metabolism
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Real-Time Polymerase Chain Reaction / veterinary

Substances

  • Glycoproteins
  • Proto-Oncogene Proteins c-bcl-2
  • RNA, Messenger
  • teleocalcin
  • Hydrogen Peroxide
  • Casp3 protein, rat
  • Caspase 3