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Effects of immunosuppressive treatment on protein expression in rat kidney
Karolina Kędzierska1, Katarzyna Sporniak-Tutak2, Krzysztof Sindrewicz2
1Department of Nephrology, Transplantology, and Internal Medicine, Pomeranian Medical University, Szczecin, Poland.
Abstract:
The structural proteins of renal tubular epithelial cells may become a target for the toxic metabolites of immunosuppressants. These metabolites can modify the properties of the proteins, thereby affecting cell function, which is a possible explanation for the mechanism of immunosuppressive agents' toxicity. In our study, we evaluated the effect of two immunosuppressive strategies on protein expression in the kidneys of Wistar rats. Fragments of the rat kidneys were homogenized after cooling in liquid nitrogen and then dissolved in lysis buffer. The protein concentration in the samples was determined using a protein assay kit, and the proteins were separated by two-dimensional electrophoresis. The obtained gels were then stained with Coomassie Brilliant Blue, and their images were analyzed to evaluate differences in protein expression. Identification of selected proteins was then performed using mass spectrometry. We found that the immunosuppressive drugs used in popular regimens induce a series of changes in protein expression in target organs. The expression of proteins involved in drug, glucose, amino acid, and lipid metabolism was pronounced. However, to a lesser extent, we also observed changes in nuclear, structural, and transport proteins' synthesis. Very slight differences were observed between the group receiving cyclosporine, mycophenolate mofetil, and glucocorticoids (CMG) and the control group. In contrast, compared to the control group, animals receiving tacrolimus, mycophenolate mofetil, and glucocorticoids (TMG) exhibited higher expression of proteins responsible for renal drug metabolism and lower expression levels of cytoplasmic actin and the major urinary protein. In the TMG group, we observed higher expression of proteins responsible for drug metabolism and a decrease in the expression of respiratory chain enzymes (thioredoxin-2) and markers of distal renal tubular damage (heart fatty acid-binding protein) compared to expression in the CMG group. The consequences of the reported changes in protein expression require further study.
Insights
Immunosuppressants alter kidney protein expression, particularly affecting drug metabolism and cellular structures. Tacrolimus-based regimens showed more significant changes than cyclosporine-based ones, impacting renal function markers.
Area of Science:
- Nephrology
- Pharmacology
- Proteomics
Background:
- Immunosuppressive drugs can exhibit nephrotoxicity through mechanisms potentially involving toxic metabolites affecting renal tubular epithelial cells.
- Understanding the impact of different immunosuppressive strategies on kidney protein expression is crucial for managing drug-induced toxicity.
Purpose of the Study:
- To investigate the effects of two distinct immunosuppressive strategies on the proteome of Wistar rat kidneys.
- To compare the protein expression profiles induced by cyclosporine, mycophenolate mofetil, and glucocorticoids (CMG) versus tacrolimus, mycophenolate mofetil, and glucocorticoids (TMG).
Main Methods:
- Kidney tissue homogenization and lysis.
- Protein concentration determination using a protein assay kit.
- Two-dimensional electrophoresis for protein separation, followed by Coomassie Brilliant Blue staining and image analysis.
- Mass spectrometry for identification of differentially expressed proteins.
Main Results:
- Both immunosuppressive regimens induced significant changes in protein expression, notably affecting proteins involved in drug, glucose, amino acid, and lipid metabolism.
- The TMG group showed increased expression of renal drug metabolism proteins and decreased expression of cytoplasmic actin and major urinary protein compared to controls.
- Compared to the CMG group, the TMG group exhibited higher expression of drug metabolism proteins, decreased respiratory chain enzymes (thioredoxin-2), and reduced markers of distal renal tubular damage (heart fatty acid-binding protein).
Conclusions:
- Immunosuppressive drug regimens significantly alter kidney proteomes, with notable impacts on metabolic pathways and structural proteins.
- The TMG regimen induced more pronounced changes in renal drug metabolism proteins and markers of tubular damage compared to the CMG regimen.
- Further research is needed to elucidate the clinical consequences of these observed protein expression changes on renal function and toxicity.
