Effects of cyclosporine A on the rat liver and kidney protein pattern, and the influence of vitamin E and C coadministration

Abstract

The effects of cyclosporine A (CsA), a potent immunosuppressive drug, were examined in rat liver and kidney samples using two-dimensional electrophoretic protein analysis. Of a total of 370 liver and 336 kidney spots analyzed, 8% (29 spots) and 6% (19 spots), respectively, showed a significant drug-induced change (p < 0.01), which was predominantly reflected in increased protein abundance (62% and 74% of the changes, respectively). Of the 48 proteins changed in either organ, 14 were most probably common to both tissues and one of these was significantly increased in both the liver and the kidney. Most of the other 13 showed similar trends (either increases or decreases) in both organs. However, the most striking drug effect seen in this study concerned an unidentified protein present only in the kidney, which completely disappeared upon CsA treatment. It was also investigated whether the drug-induced changes could be prevented by the coadministration of the radical scavengers vitamin E and C with CsA. Spots changed by the administration of the drug were classified according to three different categories, based on their response profiles in rats treated with CsA in combination with the vitamins: (i) spots which were changed by CsA as well as by CsA in combination with the vitamins (12 liver and 4 kidney spots), (ii) spots which were changed by CsA and showed an additional increase of this change by CsA plus the vitamins (no liver and 4 kidney spots), and (iii) spots which were changed by CsA but not by CsA in combination with the vitamins (8 liver and 6 kidney spots). These results showed that in both organs the vitamins were able to prevent around 30% of the effects caused by CsA, and that two-dimensional gel electrophoresis is an excellent tool to demonstrate such drug interactions at the molecular level.

Induction of the Adipose Differentiation-Related Protein in Liver of Etomoxir-Treated Rats

“…The investigation was focused on the identification of proteins which were either induced or lost in the patterns of treated rats when compared to the controls. Similar studies have been undertaken previously to investigate the liver effects of drugs such as cyclosporine A (9) or methapyrilene (10). The effects on the liver protein pattern were compared to the treatment-related histopathological changes found in the liver of the animals.…”

Protein variability in male and female Wistar rat liver proteins

“…The spot maps were displayed in the standard orientation showing high molecular mass at the top and acidic proteins to the left. In order to facilitate comparison of the Wistarlivl with the F344MST3 pattern (master based on Fisher rat liver proteins [21]), spots present in both rat strains were given identical master numbers as described previously [22]. This was achieved by matching a Wistar object pattern to a copy of the F344MST3 master followed by the deletion or addition of spots absent or specific for the Wistar rat.…”

“…1. The master pattern, which has been described previously [22], consisted of more than 1000 protein spots numbered according to a Kepler specific spot numbering system and has been standardized for the relative p l in the gel X-coordinates and for the SDS molecular weight in the gel Y-coordinates. To correct for any differences in sample loading and gel staining intensities, scaling was performed on the set of manually matched spots by setting the total protein abundance in each set equal for all gels.…”

The role of toxicoproteomics in assessing organ specific toxicity

“…An unidentified protein present only in the kidney was uniquely down-regulated [98]. A subsequent 2D gel study of kidney homogenates identified a decrease in the 28-kDa kidney protein as calbindin-D using protein microsequencing.…”