Abstract
Copper (Cu) is widely used in the swine industry to improve the growth performance of pigs. However, high doses of copper will induce cell damage and toxicity. The aim of this study was to evaluate toxicity, bioavailability, and effects on metabolic processes of varying copper sources using porcine intestinal epithelial cells (IPEC-J2) as a model. The IPEC-J2 were treated with two doses (30 and 120 μM) of CuSO4, Cu Glycine (Cu-Gly), and Cu proteinate (Cu-Pro) for 10 h, respectively. Cell damage and cellular copper metabolism were measured by the changes in cell viability, copper uptake, oxidative stress biomarkers, and gene/protein expression levels. The results showed that cell viability and ratio of reduced and oxidized glutathione (GSH/GSSG) decreased significantly in all treatment groups; intracellular copper content increased significantly in all treatment groups; total superoxide dismutase (SOD) activity increased significantly in the 120 μM exposed groups; SOD1 protein expression levels were significantly upregulated in 30 μM Cu-Pro, 120 μM Cu-Gly, and 120 μM Cu-Pro treatment groups; intracellular reactive oxygen species (ROS) generation and malondialdehyde (MDA) content increased significantly in 30 μM treatment groups and 120 μM CuSO4 treatment group. CTR1 and ATP7A gene expression were significantly downregulated in the 120 μM exposed groups. While upregulation of ATOX1 expression was observed in the presence of 120 μM Cu-Gly and Cu-Pro. ASCT2 gene expression was significantly upregulated after 120 μM Cu-Glycine and CuSO4 exposure, and PepT1 gene expression was significantly upregulated after Cu-Pro exposure. In addition, CTR1 protein expression level decreased after 120 μM CuSO4 and Cu-Gly exposure. PepT1 protein expression level was only upregulated after 120 μM Cu-Pro exposure. These findings indicated that extra copper supplementation can induce intestinal epithelial cell injury, and different forms of copper may have differing effects on cell metabolism.
Highlights
Copper is an essential trace element in animals that is beneficial to hematopoiesis, growth, metabolism, and various important physiological activities of life [1]
These findings indicated that extra copper supplementation can induce intestinal epithelial cell injury, and different forms of copper may have differing effects on cell metabolism
Copper is an essential trace element that has been involved in various biological processes
Summary
Copper is an essential trace element in animals that is beneficial to hematopoiesis, growth, metabolism, and various important physiological activities of life [1]. Studies have shown that organic metals, such as proteinate, may be absorbed attached to PepT1, a brush border membrane-bound di- and tri-peptide transporter, which has been identified in all major livestock species. This transporter is not overly specific, as molecules other than di- and tri-peptides can be transported through it. We analyzed the effects of organic and inorganic copper sources on porcine small intestinal epithelial cell line (IPEC-J2) to evaluate their effect on cell growth, oxidative stress, copper absorption efficiency, and copper metabolism, to provide data support for the precise use of copper in swine production. Three sources of copper (copper sulfate—CuSO4 , copper glycinate—Cu-Gly, and copper proteinate—Cu-Pro) were compared
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