Effect of PKC-β Signaling Pathway on Expression of MCP-1 and VCAM-1 in Different Cell Models in Response to Advanced Glycation End Products (AGEs).

Abstract

Advanced glycation end products (AGEs) are compounds classified as uremic toxins in patients with chronic kidney disease that have several pro-inflammatory effects and are implicated in the development of cardiovascular diseases. To explore the mechanisms of AGEs-endothelium interactions through the receptor for AGEs (RAGE) in the PKC-β pathway, we evaluated the production of MCP-1 and VCAM-1 in human endothelial cells (HUVECs), monocytes, and a coculture of both. AGEs were prepared by albumin glycation and characterized by absorbance and electrophoresis. The effect of AGEs on cell viability was assessed with an MTT assay. The cells were also treated with AGEs with and without a PKC-β inhibitor. MCP-1 and VCAM-1 in the cell supernatants were estimated by ELISA, and RAGE was evaluated by immunocytochemistry. AGEs exposure did not affect cell viability, but AGEs induced RAGE, MCP-1, and VCAM-1 expression in HUVECs. When HUVECs or monocytes were incubated with AGEs and a PKC-β inhibitor, MCP-1 and VCAM-1 expression significantly decreased. However, in the coculture, exposure to AGEs and a PKC-β inhibitor produced no significant effect. This study demonstrates, in vitro, the regulatory mechanisms involved in MCP-1 production in three cellular models and VCAM-1 production in HUVECs, and thus mimics the endothelial dysfunction caused by AGEs in early atherosclerosis. Such mechanisms could serve as therapeutic targets to reduce the harmful effects of AGEs in patients with chronic kidney disease.

• Authors: Lisienny C T Rempel, Alessandra B Finco, Rayana A P Maciel, Bruna Bosquetti, Larissa M Alvarenga, Wesley M Souza, Roberto Pecoits-Filho, Andréa E M Stinghen
• Journal: Toxins (Toxins (Basel)) Vol. 7 Issue 5 Pg. 1722-37 (May 14 2015) ISSN: 2072-6651 [Electronic] Switzerland
• PMID: 26008233 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• CCL2 protein, human
• Chemokine CCL2
• Glycation End Products, Advanced
• Receptor for Advanced Glycation End Products
• Vascular Cell Adhesion Molecule-1
• Protein Kinase C beta

Topics

• Cell Survival (drug effects)
• Chemokine CCL2 (metabolism)
• Coculture Techniques
• Glycation End Products, Advanced (pharmacology)
• Human Umbilical Vein Endothelial Cells (drug effects, metabolism)
• Humans
• Protein Kinase C beta (metabolism)
• Receptor for Advanced Glycation End Products (metabolism)
• Signal Transduction (drug effects)
• U937 Cells
• Vascular Cell Adhesion Molecule-1 (metabolism)