Establishment of immortalized mouse intestinal epithelial cells line and study of effects of Arg-Arg on inflammatory response.
Primary mouse intestinal epithelial cells (MIEs) are not ideal models for long-term culture in vitro and a limited amount of approximate three generations. In addition, the mechanism that arginine-arginine dipeptide (Arg-Arg) regulates mouse intestinal inflammatory response remains unknown. Therefore, the aim of this study was to establish immortal MIEs and study the effects of Arg-Arg on inflammatory response after challenging the MIEs with lipopolysaccharide (LPS) or staphylococcal enterotoxin C (rSEC). Our data showed that immortalized MIEs could be cultured over 100 generations. The immortalized MIEs showed positive reaction against cytokeratine 18 antigen, E-cadherin, and peptide transporters (Pept1) using indirect immunofluorescence. Cytokeratine 18 and Pept1 can be expressed in immortalized MIEs by immunoblotting. Fatty acid-binding proteins (FABPs) and villin known as intestinal epithelial cell functional protein were constitutively expressed in immortalized MIEs. For inflammatory response, these results showed that Arg-Arg can decrease the LPS-induced expression of IL-1β and the rSEC-induced expression of TNF-α; however, it can upregulate the LPS-induced expression of IL-6 and TNF-α and the rSEC-induced expression level of IL-1β. In addition, in the MAPK signaling pathway, pSAPK/JNK and p-Erk1/2 in LPS with Arg-Arg treatment were upregulated than that in LPS treatment. p-p38 in LPS with Arg-Arg treatment was attenuated than that in LPS treatment. pSAPK/JNK and p-p38 in rSEC with Arg-Arg treatment were enhanced than that in rSEC treatment. Conversely, p-Erk1/2 in rSEC with Arg-Arg treatment was attenuated than that in rSEC treatment. These novel findings suggest that Arg-Arg dipeptide plays an important role for regulation of the immunologic balance in mouse intestinal inflammatory response.
College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
Kang
K
Zhan
College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
Maocheng
M
Jiang
College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
Yannan
Y
Sui
College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
Kang
K
Yan
College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China.
Miao
M
Lin
College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People's Republic of China. 1751234166@qq.com.
Guoqi
G
Zhao
D000818:Animals; D004151:Dipeptides / Q000494:pharmacology*; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects; D050556:Fatty Acid-Binding Proteins / Q000235:genetics; D018507:Gene Expression Regulation, Developmental / Q000187:drug effects; D007249:Inflammation / Q000139:chemically induced / Q000235:genetics*; D007422:Intestines / Q000166:cytology* / Q000187:drug effects / Q000254:growth & development; D008070:Lipopolysaccharides / Q000633:toxicity; D051379:Mice
D004151:Dipeptides; D050556:Fatty Acid-Binding Proteins; D008070:Lipopolysaccharides; C076685:arginylarginine
Arg-Arg; Cytokeratine 18; Immortalize; Mouse intestinal epithelial cells; Villin