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Butyrate upregulates proglucagon mRNA abundance through activation of the proglucagon gene promoter in NCI H716 cells
Author(s) -
Woodard Jennifer N.,
Tappenden Kelly A.
Publication year - 2008
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.22.2_supplement.135
Subject(s) - proglucagon , butyrate , biology , messenger rna , microbiology and biotechnology , endocrinology , medicine , chemistry , gene , glucagon like peptide 1 , biochemistry , fermentation , type 2 diabetes , diabetes mellitus
The proglucagon gene encodes for 5 peptides, including the intestinotrophic hormone glucagon‐like peptide 2 (GLP‐2). Previous in vivo work has shown butyrate to stimulate intestinal adaptation, increase plasma GLP‐2 and intestinal proglucagon mRNA levels. The aim of this study was to examine the transcriptional regulation of proglucagon by butyrate. Using NCI H716 cells, we hypothesized that proglucagon mRNA abundance increases in response to butyrate due to activation of the proglucagon promoter. NCI H716 cells received 1 of 6 doses of butyrate (0–20 mM) and were harvested at 4 time points (0.5–4 hours). Quantitative real‐time PCR revealed proglucagon mRNA abundance increased (p<0.05) with 7.5 mM butyrate following 2 hours of treatment (2.16‐fold increase compared to 0 mM). 2500 sequence‐confirmed base pairs of the human proglucagon promoter were isolated, cloned into the pGL4 reporter vector, and transiently transfected into NCI H716 cells. Following treatment with 7.5 mM butyrate, luciferase activity, as driven by the proglucagon promoter, was increased (p<0.05; 1.8‐fold increase compared to controls). In summary, we have shown butyrate to upregulate proglucagon mRNA abundance through an activation of the proglucagon gene promoter, thereby furthering our understanding of the molecular mechanisms wherein butyrate and GLP‐2 interact during intestinal adaptation. Supported by NIDDK 1 R01 DK 57682.