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Cyclosporine induces different responses in human epithelial, endothelial and fibroblast cell cultures
C Esposito1, A Fornoni, F Cornacchia
1Division of Nephrology and Dialysis, IRCCS Policlinico San Matteo, University of Pavia, Italy. ciroe@pop.systemy.it
Background:
Nephrotoxicity, accelerated atherosclerosis, and graft vascular disease are common complications of cyclosporine long-term treatment characterized by a wide disruption of organ architecture with increased interstitial areas and accumulation of extracellular matrix (ECM). How cyclosporine induces these changes is not clear, but it is conceivable that they are the sum of changes induced at the cell level.
Methods:
We studied the effects of cyclosporine on human endothelial (HEC), epithelial (HK-2), and fibroblast (MRC5) cells. Cell proliferation was evaluated by cell counting, apoptosis and collagen production by enzyme-linked immunosorbent assay, and nitric oxide by measuring the concentration of nitrite/nitrate in the cell supernatant. (alpha1)I and (alpha2)IV collagen, matrix metalloprotease-9 (MMP9), and tissue inhibitors of metalloprotease-1 (TIMP-1) mRNA levels were measured by reverse transcription-polymerase chain reaction. Proteolytic activity was evaluated by zymography.
Results:
Cyclosporine showed a marked antiproliferative and proapoptotic effect on endothelial and epithelial cells. Fibroblast growth was not affected by cyclosporine. Nitric oxide was up-regulated by cyclosporine in epithelial cells and fibroblasts but not in endothelial cells. (alpha1)I and (alpha2)IV collagen synthesis was increased in cyclosporine-treated endothelial and epithelial cells, respectively. Proteolytic activity was increased in endothelial and epithelial cells. TIMP-1 mRNA was up-regulated by cyclosporine in fibroblasts.
Conclusions:
Our results demonstrate that cyclosporine exhibits an antiproliferative effect on endothelial and epithelial cells. This effect is associated with induction of apoptosis probably via nitric oxide up-regulation in epithelial cell cultures. Cyclosporine treatment induces ECM accumulation by increasing collagen synthesis in endothelial and epithelial cells and reducing its degradation by up-regulating TIMP-1 expression in fibroblasts. We conclude that cyclosporine affects cell types differently and that the disruption of organ architecture is the result of multiple effects at the cell level.
Insights
Cyclosporine hinders endothelial and epithelial cell growth, inducing apoptosis and extracellular matrix accumulation. These cellular changes, varying by cell type, contribute to organ damage from long-term cyclosporine use.
Area of Science:
- Cell Biology
- Immunology
- Pharmacology
Background:
- Long-term cyclosporine treatment can cause nephrotoxicity, accelerated atherosclerosis, and graft vascular disease.
- These complications involve organ architecture disruption and extracellular matrix (ECM) accumulation.
- The cellular mechanisms underlying cyclosporine-induced organ damage are not fully understood.
Purpose of the Study:
- To investigate the effects of cyclosporine on human endothelial cells (HEC), epithelial cells (HK-2), and fibroblasts (MRC5).
- To elucidate the cellular mechanisms by which cyclosporine may induce organ damage.
Main Methods:
- Assessed cell proliferation, apoptosis, and nitric oxide production.
- Quantified collagen synthesis and mRNA levels of collagen, MMP9, and TIMP-1.
- Evaluated proteolytic activity using zymography.
Main Results:
- Cyclosporine exhibited antiproliferative and proapoptotic effects on HEC and HK-2 cells, but not MRC5 cells.
- Nitric oxide was upregulated in HK-2 and MRC5 cells, but not HEC.
- Increased collagen synthesis and proteolytic activity were observed in HEC and HK-2 cells; TIMP-1 mRNA was upregulated in MRC5 cells.
Conclusions:
- Cyclosporine differentially affects cell types, with antiproliferative and proapoptotic effects on endothelial and epithelial cells.
- ECM accumulation is induced by increased collagen synthesis and reduced degradation via TIMP-1 upregulation in fibroblasts.
- The observed cellular effects provide insight into cyclosporine-induced organ architecture disruption.