Cyclic AMP reverses radiocontrast media-induced apoptosis in LLC-PK1 cells by activating a kinase/PI3 kinase

Takahisa Yano, Yoshinori Itoh, Toshiaki Sendo, Toshio Kubota, Ryozo Oishi

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Background. Radiographic contrast material is one of agents that are prone to cause nephropathy, although little is known about cellular mechanisms underlying contrast media-induced renal failure. The present study was designed to determine the role of caspase in contrast media-induced renal injury. The modulation by cyclic adenosine monophosphate (cAMP) of cell injury was subsequently examined. Methods. LLC-PK1 cells (a proximal renal tubular cell line of porcine origin) were exposed to diverse contrast media for 30 minutes followed by incubation for 24 hours in normal medium. Cell viability was assessed by mitochondrial enzyme activity and propidium iodide stain. Apoptosis was determined by DNA electrophoresis and annexin V stain. Caspase activity was measured fluorometrically. The mRNA for bax and bcl-2 was determined by reverse transcription-polymerase chain reaction (RT-PCR). Results. Iodinated and magnetic resonance contrast media reduced cell viability due to apoptosis. The cell damage induced by a non-ionic contrast medium ioversol was inhibited by specific inhibitors for caspase-3 and -9 but not caspase-8. Ioversol enhanced the activities of caspase-3 and -9, but to a lesser extent, caspase-8. The bax mRNA was enhanced, while bcl-2 mRNA was reduced, after exposure to ioversol. All of these actions of ioversol were reversed by dibutyl cAMP in the manner sensitive to a protein kinase A inhibitor H89 and a phosphatidylinositol 3 (PI3) kinase inhibitor wortmannin. Conclusion. We demonstrated for the first time that cAMP reversed caspase-dependent apoptotic renal cell damage caused by contrast media. Both protein kinase A and PI3 kinase might be involved in protective effect of cAMP.

Original languageEnglish
Pages (from-to)2052-2063
Number of pages12
JournalKidney International
Volume64
Issue number6
DOIs
Publication statusPublished - Dec 2003
Externally publishedYes

Fingerprint

Phosphatidylinositol 3-Kinase
LLC-PK1 Cells
ioversol
Cyclic AMP
Contrast Media
Phosphotransferases
Apoptosis
Caspases
Caspase 9
Caspase 8
Cyclic AMP-Dependent Protein Kinases
Kidney
Caspase 3
Messenger RNA
Cell Survival
Coloring Agents
Propidium
Annexin A5
Wounds and Injuries
Protein Kinase Inhibitors

Keywords

  • Apoptosis
  • Bax
  • bcl-2
  • cAMP
  • Caspase
  • Contrast media
  • PI3 kinase
  • PKA
  • Renal tubular cells

ASJC Scopus subject areas

  • Nephrology

Cite this

Cyclic AMP reverses radiocontrast media-induced apoptosis in LLC-PK1 cells by activating a kinase/PI3 kinase. / Yano, Takahisa; Itoh, Yoshinori; Sendo, Toshiaki; Kubota, Toshio; Oishi, Ryozo.

In: Kidney International, Vol. 64, No. 6, 12.2003, p. 2052-2063.

Research output: Contribution to journalArticle

Yano, Takahisa ; Itoh, Yoshinori ; Sendo, Toshiaki ; Kubota, Toshio ; Oishi, Ryozo. / Cyclic AMP reverses radiocontrast media-induced apoptosis in LLC-PK1 cells by activating a kinase/PI3 kinase. In: Kidney International. 2003 ; Vol. 64, No. 6. pp. 2052-2063.
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AU - Kubota, Toshio

AU - Oishi, Ryozo

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N2 - Background. Radiographic contrast material is one of agents that are prone to cause nephropathy, although little is known about cellular mechanisms underlying contrast media-induced renal failure. The present study was designed to determine the role of caspase in contrast media-induced renal injury. The modulation by cyclic adenosine monophosphate (cAMP) of cell injury was subsequently examined. Methods. LLC-PK1 cells (a proximal renal tubular cell line of porcine origin) were exposed to diverse contrast media for 30 minutes followed by incubation for 24 hours in normal medium. Cell viability was assessed by mitochondrial enzyme activity and propidium iodide stain. Apoptosis was determined by DNA electrophoresis and annexin V stain. Caspase activity was measured fluorometrically. The mRNA for bax and bcl-2 was determined by reverse transcription-polymerase chain reaction (RT-PCR). Results. Iodinated and magnetic resonance contrast media reduced cell viability due to apoptosis. The cell damage induced by a non-ionic contrast medium ioversol was inhibited by specific inhibitors for caspase-3 and -9 but not caspase-8. Ioversol enhanced the activities of caspase-3 and -9, but to a lesser extent, caspase-8. The bax mRNA was enhanced, while bcl-2 mRNA was reduced, after exposure to ioversol. All of these actions of ioversol were reversed by dibutyl cAMP in the manner sensitive to a protein kinase A inhibitor H89 and a phosphatidylinositol 3 (PI3) kinase inhibitor wortmannin. Conclusion. We demonstrated for the first time that cAMP reversed caspase-dependent apoptotic renal cell damage caused by contrast media. Both protein kinase A and PI3 kinase might be involved in protective effect of cAMP.

AB - Background. Radiographic contrast material is one of agents that are prone to cause nephropathy, although little is known about cellular mechanisms underlying contrast media-induced renal failure. The present study was designed to determine the role of caspase in contrast media-induced renal injury. The modulation by cyclic adenosine monophosphate (cAMP) of cell injury was subsequently examined. Methods. LLC-PK1 cells (a proximal renal tubular cell line of porcine origin) were exposed to diverse contrast media for 30 minutes followed by incubation for 24 hours in normal medium. Cell viability was assessed by mitochondrial enzyme activity and propidium iodide stain. Apoptosis was determined by DNA electrophoresis and annexin V stain. Caspase activity was measured fluorometrically. The mRNA for bax and bcl-2 was determined by reverse transcription-polymerase chain reaction (RT-PCR). Results. Iodinated and magnetic resonance contrast media reduced cell viability due to apoptosis. The cell damage induced by a non-ionic contrast medium ioversol was inhibited by specific inhibitors for caspase-3 and -9 but not caspase-8. Ioversol enhanced the activities of caspase-3 and -9, but to a lesser extent, caspase-8. The bax mRNA was enhanced, while bcl-2 mRNA was reduced, after exposure to ioversol. All of these actions of ioversol were reversed by dibutyl cAMP in the manner sensitive to a protein kinase A inhibitor H89 and a phosphatidylinositol 3 (PI3) kinase inhibitor wortmannin. Conclusion. We demonstrated for the first time that cAMP reversed caspase-dependent apoptotic renal cell damage caused by contrast media. Both protein kinase A and PI3 kinase might be involved in protective effect of cAMP.

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