Premium
Glucagon‐Like Peptide‐1 (GLP‐1) Receptor Expression on Human Umbilical Vein Endothelial Cells (HUVEC) in Response to Cellular Activation
Author(s) -
Noormohamed Nadia,
Kiel Dan
Publication year - 2016
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.30.1_supplement.1269.6
Subject(s) - receptor , nitric oxide , umbilical vein , microbiology and biotechnology , cell surface receptor , chemistry , intracellular , endothelial stem cell , tumor necrosis factor alpha , glucagon like peptide 1 receptor , nitric oxide synthase , endocrinology , biology , biochemistry , in vitro , agonist
Purpose The Glucagon‐Like Peptide‐1 (GLP‐1) receptor is the target for certain anti‐diabetic medications. The GLP‐1 receptor is a cell‐surface receptor that elicits a number of responses upon activation leading to reduced blood glucose and various other effects. A number of cell‐surface proteins have been shown to change expression level in response to various stimuli, such as nitric oxide and cytokines. Alterations in the level of cell‐surface receptors can increase or decrease responses through that receptor. The purpose of this study is to measure GLP‐1 receptor expression on human umbilical vein endothelial cells (HUVEC) in response to cellular activation by various compounds. Methods Nitric oxide and tumor necrosis factor alpha (TNF‐alpha) have been shown to induce changes in levels of cell‐surface receptors. Puerarin is an isoflavone found in a number of plants and herbs that stimulates endothelial nitric oxide synthase, leading to the generation of intracellular nitric oxide. Nitric oxide activates intracellular enzymes, such as guanylate cyclase, leading to the generation of second messengers. TNF‐alpha is a cytokine that increases transcription of various genes including those for cell surface adhesion molecules involved in the inflammatory process. HUVEC monolayers were treated with puerarin, TNF‐alpha, or vehicle. After 24 hours of treatment, cells were harvested. Equivalent amounts of cellular homogenates were separated via electrophoresis, and proteins transferred to a membrane. The membrane was probed with an antibody directed against the GLP‐1 receptor. Visualization of the band corresponding to the GLP‐1 receptor was achieved using chemiluminescence, and quantified using densitometry. Neither of the treatments produced large changes in GLP‐1 receptor expression. GLP‐1 receptor expression on HUVEC may be less responsive to cellular stimulation compared to other proteins on HUVEC.