Proteomic Analyses of Contrast Media-Treated Mesangial Cell

Abstract

Contrast Media is used in angiography, urography and tomography. It has been reported that contrast media nephrotoxicity incidence is 50% higher in patients with diabetic nephropathy undergoing coronary angiography and approximately 15% of these patients developed renal failure. Proteomic analyses are a promising tool to study renal pathophysiology. In the future identification of biomarkers in renal diseases will develop therapeutic targets that will decrease the damage of acute renal failure. To identify proteins that were up or down-regulated in immortalized mesangial cells in a culture in the presence of contrast media we evaluated three groups: control, manitol used as control of osmolarity and Hexabrix TM . The cell homogenate from each group was submitted to the 2D-PAGE analysis, and proteins were identified by MALDI-TOF mass spectrometry. Some proteins were expressed only in the Hexabrix group (Proliferating Cell Nuclear Antigen and Masp 1); others were up-regulated in the Manitol group (Translationally Controlled Tumor Protein), a protein was expressed only in the Control and Manitol (Annexin A3) groups, and we identified proteins that were expressed in all three groups (Heat shock proteins 27 and 84, protein disulfide isomerase A3 precursor and Beta-actin 1). We believe that other substances present in Hexabrix, possibly the iodine, could be the regulatory factor of these proteins and the Hexabrix could be responsible for the nephrotoxicity observed in these cells.

Proteomic profiling of the rat hypothalamus

“…The selected spots were automatically excised, destained and digested, using Xcise TM spot picker (Shimadzu Biotech, Japan), as reported elsewhere [40]. Briefly, excised spots were destained in 100 mM ammonium bicarbonate and 50% acetonitrile (1:1) and digestion was performed in 30 μL of 10 ng/μL trypsin (Sigma Aldrich, Germany) in 25 mM ammonium bicarbonate, overnight at 34°C.…”