J Korean Surg Soc.  2009 Mar;76(3):135-143. 10.4174/jkss.2009.76.3.135.

Preventive Effect of Pentoxifylline on Cyclosporine A-Induced Collagen Synthesis in Calf Pulmonary Artery Endothelial Cells

• Kim SE
• Sung YH
• Shin MS
• Kim CJ
• Park JH
• Lee BJ
• Yi JW
• Han SY

• KMID: 2096534
• DOI: http://doi.org/10.4174/jkss.2009.76.3.135

Abstract

PURPOSE
Cyclosporine A (CsA) is a potent immunosuppressive agent, and it has been used to prevent rejection of transplanted organs and to treat autoimmune diseases. Many side effects of CsA, including various types of endothelial dysfunction, have been reported. Pentoxifylline (PTX) is a non-selective phosphodiesterase inhibitor that is used for the treatment of peripheral vascular diseases. METHODS: We investigated the effect of CsA on collagen synthesis and clarified whether PTX has protective effects against CsA-induced arterial vasculopathy using calf pulmonary artery endothelial cells. This study was carried out using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcription- polymerase chain reaction (RT-PCR), Western blot analysis, nitric oxide (NO) detection, and cyclic guanosine monophosphate (cGMP) enzyme immunoassay. RESULTS: CsA treatment significantly increased the expression of collagen type I mRNA and protein and decreased the production of NO and cGMP. However, pre-treatment with PTX exerted anticollagen effect by suppressing the CsA-induced formation of collagen, but this effect of PTX was not modulated by NO and cGMP. CONCLUSION: Based on the present results, it is expected that PTX may have a protective effect against CsA-induced arterial vasculopathy, although the mechanism of PTX needs to be clarified in future studies.

Keyword

Cyclosporine A; Pentoxifylline; Collagen type I; Cyclic GMP; Nitric oxide

MeSH Terms

Autoimmune Diseases
Blotting, Western
Collagen
Collagen Type I
Cyclic GMP
Cyclosporine
Endothelial Cells
Guanosine Monophosphate
Immunoenzyme Techniques
Nitric Oxide
Pentoxifylline
Peripheral Vascular Diseases
Polymerase Chain Reaction
Pulmonary Artery
Rejection (Psychology)
RNA, Messenger
Tetrazolium Salts
Thiazoles
Transplants
Collagen
Collagen Type I
Cyclic GMP
Cyclosporine
Guanosine Monophosphate
Nitric Oxide
Pentoxifylline
RNA, Messenger
Tetrazolium Salts
Thiazoles

Figure

• 

  Fig. 1 Cytotoxic effect of cyclosporine A (CsA) on CPAE cells. (A) Control group. (B) 0.001 µM cyclosporine A (CsA)-treated group. (C) 0.01 µM CsA-treated group. (D) 0.1 µM CsA-treated group. (E) 1 µM CsA-treated group. (F) 10 µM CsA-treated group. The results are presented as the mean±standard error of the mean (SEM). *P<0.05 compared to the control group.

• 

  Fig. 2 Cytotoxic effects of pentoxifylline (PTX) on CPAE cells. (A) Control group. (B) 5 µM pentoxifylline (PTX)-treated group. (C) 10 µM PTX-treated group. (D) 50 µM PTX-treated group. (E) 100 µM PTX-treated group. (F) 500 µM PTX-treated group. (G) 1,000 µM PTX-treated group. The results are presented as the mean±standard error of the mean (SEM). *P<0.05 compared to the control group.

• 

  Fig. 3 Effects of cyclosporine A (CsA) and pentoxifylline (PTX) on the expression of collagen type I mRNA. (A) Control group. (B) 1 µM cyclosporine A (CsA)-treated group. (C) 1 µM CsA- and 100 µM pentoxifylline (PTX)-treated group. (D) 1 µM CsA- and 1,000 µM PTX-treated group. The results are presented as the mean±standard error of the mean (SEM). *P<0.05 compared to the control group. †P<0.05 compared to the 1 µM CsA-treated group.

• 

  Fig. 4 Effects of cyclosporine A (CsA) and pentoxifylline (PTX) on the expression of collagen type I protein. (A) Control group. (B) 1 µM cyclosporine A (CsA)-treated group. (C) 1 µM CsA- and 100 µM pentoxifylline (PTX)-treated group. (D) 1 µM CsA- and 1,000 µM PTX-treated group. The results are presented as the mean±standard error of the mean (SEM). *P<0.05 compared to the control group. †P<0.05 compared to the 1 µM CsA-treated group.

• 

  Fig. 5 Effects of cyclosporine A (CsA) and pentoxifylline (PTX) on nitric oxide (NO) production. (A) Control group. (B) 1 µM cyclosporine A (CsA)-treated group. (C) 1 µM CsA- and 100 µM pentoxifylline (PTX)-treated group. (D) 1 µM CsA- and 1,000 µM PTX-treated group. The results are presented as the mean†standard error of the mean (SEM). *P<0.05 compared to the control group. †P<0.05 compared to the 1 µM CsA-treated group.

• 

  Fig. 6 Effects of cyclosporine A (CsA) and pentoxifylline (PTX) on cyclic guanosine monophosphate (cGMP) production. (A) Control group. (B) 1 µM cyclosporine A (CsA)-treated group. (C) 1 µM CsA- and 100 µM pentoxifylline (PTX)-treated group. (D) 1 µM CsA- and 1,000 µM PTX-treated group. The results are presented as the mean±standard error of the mean (SEM). *P<0.05 compared to the control group. †P<0.05 compared to the 1 µM CsA-treated group.

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