Session: MON 649-675-Central Regulation of Appetite & Feeding/GI Regulatory Peptides
Bench to Bedside
Poster Board MON-667
Using the Caco-2 human intestinal epithelial cells we investigated the effect of specific macronutrients on PYY and DPPIV gene expression. Cells were stimulated for 24 hours with 1.59 mM L-Leucine, 2.39 mM L-Arginine, L-Leu+L-Arg, Non Essential Amino Acids (NEAA) x 10 concentration and 20 nM glucose. The expression of DPP-IV and PYY1-36 genes was determined by SYBR-green real time PCR. Cell culture supernatant PYY 3-36 protein was measured using a human enzyme immunoassay (EIA) kit. The protein concentration was standardised against total protein content quantified using bicinchoninic assay (BCA). Caco-2 cells were also incubated for 3 hours at varying concentrations (20, 50, 100, 200 and 500 nM) of exogenous PYY1-36. PYY3-36 protein was measured in cell culture supernatant.
Our results showed no significant change to the DPP-IV and PYY1-36 gene and PYY3-36 protein expression on stimulation with either L-Leu or L-Arg in isolation. However, addition of both L-Leu+L-Arg in combination resulted in a 1.5 fold induction of DPP-IV (P=0.017) gene and 1.8 fold up-regulation of PYY1-36 (P=0.019) gene expression. Similarly, the cellular PYY3-36 protein was significantly up-regulated when compared to control (P = 0.04) on stimulation with both L-Leu+L-Arg. Glucose treatment also caused a 1.4 up-regulation of DPPIV (P=0.041) and a 3.3 up-regulation of PYY1-36 (P<0.0001) genes. This gene induction was reflected in a significant increase in cellular PYY3-36 protein (P=0.01). Furthermore, incubation of Caco-2 cells with varying concentrations of exogenous PYY1-36 resulted in a linear increased concentration of PYY3-36 (R2=0.32, P<0.0001).These data show for the first time that PYY gene is expressed in the human gut epithelial cell line- Caco-2 and that stimulation with specific macronutrients leads to an increase in PYY3-36 gene expression and protein release as well as an up regulation of DPPIV gene.
Nothing to Disclose: MGZ, AK, DR
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