The Appearance of a Leptin Effect on Glucose Absorption in Caco2 Cells Depends on Their Differentiation Level
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S. Karger AG
Abstract
Backdround/Aims: The aim of this work was to study the effect and mechanism of action of leptin added apically, on glucose absorption, using Caco-2 cells as a model. Methods: Cells were grown on inserts and treated with leptin, at different time points after confluence. Radiolabelled glucose was added to the upper chamber and samples from the lower chamber were collected and assayed for radioactivity. Results: Glucose absorption increased with an increase in the level of differentiation and was associated with an increase in the protein expression level of glucose transporters. Leptin reduced glucose absorption only by day 16 after confluence, the time at which apical leptin receptors started appearing. This inhibitory effect became higher the longer the post confluence period, and was prominent on day 23. The hormone effect was found to be mediated via a decrease in the number of glucose transporters (SGLT1 and GLUT2) and a decrease in the activity of the Na + /K + ATPase which was assayed by measuring the amount of inorganic phosphate liberated in presence and absence of enzyme activators. Conclusion: It was concluded that by day 23 post confluence, Caco-2 cells are differentiated and are appropriate to use as a model for intestinal transport studies. © 2015 S. Karger AG, Basel.
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Caco-2, Glut-2, Leptin, Leptin receptors, Na + /k + atpase, Sglt1, Caco-2 cells, Cell differentiation, Gene expression regulation, Glucose, Glucose transporter type 2, Humans, Intestinal absorption, Models, biological, Sodium-glucose transporter 1, Sodium-potassium-exchanging atpase, Adenosine triphosphatase (potassium sodium), Glucose transporter, Glucose transporter 2, Leptin receptor, Phosphate, Sodium glucose cotransporter 1, Slc2a2 protein, human, Slc5a1 protein, human, Article, Basolateral membrane, Caco 2 cell line, Controlled study, Enzyme activity, Enzyme assay, Glucose absorption, Human, Human cell, Priority journal, Protein expression, Biological model, Drug effects, Intestine absorption, Metabolism