Studies Of The Regulation Of Glutamine Dipeptide On Cell Renewal And Protein Metabolism In Porcine Intestinal Epithelial Cell

The objective of this thesis was to preliminarily explore Glutamine dipeptide to regulate the porcine intestinal epithelial cell renewal and the Regulatory mechanism of protein metabolism. Use the way of culturing porcine intestinal epithelial cell (IPEC-J2) in vitro, with an equal dose concentration (2.5mM) of Ala-Gln and Gly-Gln insteading of Gin in DMEM-F12 medium.Take methods and techniques of Edu incorporation, Flow cytometric analysis, Isotopic tracing, RT-PCR and Western Blot to research Glutamine dipeptide on the regulatory role of cell proliferation, small peptide transporter's expression, protein turnover and the related signaling pathways.The results show:Compared with Gin, Ala-Gln has not significant effect on cell proliferation and cell cycle, Gly-Gln inhibited cell proliferation; Ala-Gln promotes small peptide transporter PepT1 gene expression levels, but has not significant effect on its transcription factor Spl; Ala-Gln-treated group and Gin-treated group have not significant effect on cellular protein turnover, but Gly-Gln significantly inhibit protein synthesis and promote protein degradation; Ala-Gln-treated cells mTOR phosphorylation levels,4EBP1 phosphorylation levels and S6K1 protein expression levels were significantly higher than Gln-treated cells, but these levels of Gly-Gln-treated cells were significantly lower than other treated cells; Ala-Gln-treated cells UBE3B mRNA expression levels were significantly higher than Gln-treated cells, but Gly-Gln-treated and Gln-treated cells have not significant effect on UBE3B mRNA expression levels.These results suggest that Ala-Gln can replace of Gin to play a role on promoting cell proliferation and protein synthesis culturing intestinal epithelial cells in vitro.