1.Stanniocalcin-1 protects bovine intestinal epithelial cells from oxidative stress-induced damage.
Li Ming WU ; Rui GUO ; Lin HUI ; Yong Gang YE ; Jing Mei XIANG ; Chun Yun WAN ; Miao ZOU ; Rui MA ; Xiao Zhuan SUN ; Shi Jin YANG ; Ding Zong GUO
Journal of Veterinary Science 2014;15(4):475-483
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 microM 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.
Animals
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Animals, Newborn
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Blotting, Western/veterinary
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Caspase 3/*genetics/metabolism
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Cattle
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Cattle Diseases/etiology/*genetics/metabolism
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Duodenum/metabolism
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Enteritis/etiology/genetics/metabolism/*veterinary
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Epithelial Cells/metabolism
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*Gene Expression Regulation
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Glycoproteins/*genetics/metabolism
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Hydrogen Peroxide/pharmacology
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Male
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Proto-Oncogene Proteins c-bcl-2/*genetics/metabolism
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RNA, Messenger/genetics/metabolism
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Real-Time Polymerase Chain Reaction/veterinary