Cyclosporine induces different responses in human epithelial, endothelial and fibroblast cell cultures

Ciro Esposito, Alessia Fornoni, Flavia Cornacchia, Nicoletta Bellotti, Gianluca Fasoli, Annalisa Foschi, Iolanda Mazzucchelli, Tiziana Mazzullo, Luca Semeraro, Antonio Dal Canton

Research output: Contribution to journalArticle

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Abstract

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 call counting, apoptosis and collagen production by enzyme-linked immunosorbent assay, and nitric oxide by measuring the concentration of nitrite/nitrate in the cell supernatant. (α1)I and (α2)IV collagen, matrix metallapratease-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. (α1)I and (α2)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 upregulated 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.

Original languageEnglish
Pages (from-to)123-130
Number of pages8
JournalKidney International
Volume58
Issue number1
DOIs
StatePublished - Jul 15 2000
Externally publishedYes

Fingerprint

Cyclosporine
Endothelial Cells
Cell Culture Techniques
Fibroblasts
Epithelial Cells
Collagen
Nitric Oxide
Metalloproteases
Extracellular Matrix
Apoptosis
Messenger RNA
Tissue Inhibitor of Metalloproteinase-1
Nitrites
Vascular Diseases
Nitrates
Reverse Transcription
Atherosclerosis
Up-Regulation
Enzyme-Linked Immunosorbent Assay
Cell Proliferation

Keywords

  • Apoptosis
  • Cell proliferation
  • Matrix turnover
  • Nitric oxide
  • Transplantation

ASJC Scopus subject areas

  • Nephrology

Cite this

Cyclosporine induces different responses in human epithelial, endothelial and fibroblast cell cultures. / Esposito, Ciro; Fornoni, Alessia; Cornacchia, Flavia; Bellotti, Nicoletta; Fasoli, Gianluca; Foschi, Annalisa; Mazzucchelli, Iolanda; Mazzullo, Tiziana; Semeraro, Luca; Canton, Antonio Dal.

In: Kidney International, Vol. 58, No. 1, 15.07.2000, p. 123-130.

Research output: Contribution to journalArticle

Esposito, C, Fornoni, A, Cornacchia, F, Bellotti, N, Fasoli, G, Foschi, A, Mazzucchelli, I, Mazzullo, T, Semeraro, L & Canton, AD 2000, 'Cyclosporine induces different responses in human epithelial, endothelial and fibroblast cell cultures', Kidney International, vol. 58, no. 1, pp. 123-130. https://doi.org/10.1046/j.1523-1755.2000.00147.x
Esposito, Ciro ; Fornoni, Alessia ; Cornacchia, Flavia ; Bellotti, Nicoletta ; Fasoli, Gianluca ; Foschi, Annalisa ; Mazzucchelli, Iolanda ; Mazzullo, Tiziana ; Semeraro, Luca ; Canton, Antonio Dal. / Cyclosporine induces different responses in human epithelial, endothelial and fibroblast cell cultures. In: Kidney International. 2000 ; Vol. 58, No. 1. pp. 123-130.
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AU - Esposito, Ciro

AU - Fornoni, Alessia

AU - Cornacchia, Flavia

AU - Bellotti, Nicoletta

AU - Fasoli, Gianluca

AU - Foschi, Annalisa

AU - Mazzucchelli, Iolanda

AU - Mazzullo, Tiziana

AU - Semeraro, Luca

AU - Canton, Antonio Dal

PY - 2000/7/15

Y1 - 2000/7/15

N2 - 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 call counting, apoptosis and collagen production by enzyme-linked immunosorbent assay, and nitric oxide by measuring the concentration of nitrite/nitrate in the cell supernatant. (α1)I and (α2)IV collagen, matrix metallapratease-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. (α1)I and (α2)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 upregulated 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.

AB - 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 call counting, apoptosis and collagen production by enzyme-linked immunosorbent assay, and nitric oxide by measuring the concentration of nitrite/nitrate in the cell supernatant. (α1)I and (α2)IV collagen, matrix metallapratease-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. (α1)I and (α2)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 upregulated 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.

KW - Apoptosis

KW - Cell proliferation

KW - Matrix turnover

KW - Nitric oxide

KW - Transplantation

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