Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells

Mai Szu Wu, Chih Wei Yang, Marcelle Bens, Hsiao Mei Yu, Jeng Yi Huang, Chin Herng Wu, Chiu Ching Huang, Alain Vandewalle

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Background. Nitric oxide (NO) has been shown to play a role in cyclosporin (CsA) nephrotoxicity, but its mechanism of action is still unclear. As inducible NO synthase (iNOS) mRNA has been found to be expressed in rat medullary thick ascending limb (mTAL) cells, we investigated the effects of CsA on NO production in a model of mouse cultured mTAL cells. Materials and methods. The experiments were carried out on sub-cultured cells derived from isolated mTAL microdissected from the kidney of C57BL/6 mice. The identification of the iNOS mRNA in mTAL microdissected segment and cultured cell was confirmed by RT-PCR and RsaI digestion. Nitrite (NO2 -) released by mTAL cells was determined using the modified Griess reagent method and taken as an index of nitric oxide production. The cultured cells were treated with various concentrations of CsA and different signal transduction regulators to assess the effect and possible pathway(s) of action of CsA on NO production in mTAL cells. Results. The basal production of NO by mTAL cells increased by 1.8-fold following incubation with bacterial lipopolysaccaride (LPS). Both aminoguanidine and L-NAME inhibited NO production. CsA (10-300 ng/ml) also inhibited NO production in a dose-dependent manner and prevented its increase induced by LPS. Phorbol 12-myristate 13-acetate (PMA), a PKC stimulator, enhanced slightly the production of NO under basal conditions and prevented the inhibitory action of CsA on NO production. These results suggest that the NO secreted by mouse cultured mTAL cells is dependent on the PKC pathway. Conclusion. These results show that CsA may downregulate the production of NO by cultured mTAL cells expressing iNOS mRNA and that the PKC pathway is involved in this process.

Original languageEnglish
Pages (from-to)2814-2820
Number of pages7
JournalNephrology Dialysis Transplantation
Volume13
Issue number11
Publication statusPublished - Nov 1998
Externally publishedYes

Fingerprint

Cyclosporine
Cultured Cells
Nitric Oxide
Extremities
Nitric Oxide Synthase
Messenger RNA
NG-Nitroarginine Methyl Ester
Nitric Oxide Synthase Type II
Nitrites
Inbred C57BL Mouse
Digestion
Signal Transduction
Acetates
Down-Regulation
Kidney
Polymerase Chain Reaction

Keywords

  • Cultured cells
  • Cyclosporin
  • Kidney
  • Nephrotoxicity
  • Nitric oxide
  • Thick ascending limbs

ASJC Scopus subject areas

  • Nephrology
  • Transplantation

Cite this

Wu, M. S., Yang, C. W., Bens, M., Yu, H. M., Huang, J. Y., Wu, C. H., ... Vandewalle, A. (1998). Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells. Nephrology Dialysis Transplantation, 13(11), 2814-2820.

Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells. / Wu, Mai Szu; Yang, Chih Wei; Bens, Marcelle; Yu, Hsiao Mei; Huang, Jeng Yi; Wu, Chin Herng; Huang, Chiu Ching; Vandewalle, Alain.

In: Nephrology Dialysis Transplantation, Vol. 13, No. 11, 11.1998, p. 2814-2820.

Research output: Contribution to journalArticle

Wu, MS, Yang, CW, Bens, M, Yu, HM, Huang, JY, Wu, CH, Huang, CC & Vandewalle, A 1998, 'Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells', Nephrology Dialysis Transplantation, vol. 13, no. 11, pp. 2814-2820.
Wu, Mai Szu ; Yang, Chih Wei ; Bens, Marcelle ; Yu, Hsiao Mei ; Huang, Jeng Yi ; Wu, Chin Herng ; Huang, Chiu Ching ; Vandewalle, Alain. / Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells. In: Nephrology Dialysis Transplantation. 1998 ; Vol. 13, No. 11. pp. 2814-2820.
@article{9fac403da47042f5b562ce361515c422,
title = "Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells",
abstract = "Background. Nitric oxide (NO) has been shown to play a role in cyclosporin (CsA) nephrotoxicity, but its mechanism of action is still unclear. As inducible NO synthase (iNOS) mRNA has been found to be expressed in rat medullary thick ascending limb (mTAL) cells, we investigated the effects of CsA on NO production in a model of mouse cultured mTAL cells. Materials and methods. The experiments were carried out on sub-cultured cells derived from isolated mTAL microdissected from the kidney of C57BL/6 mice. The identification of the iNOS mRNA in mTAL microdissected segment and cultured cell was confirmed by RT-PCR and RsaI digestion. Nitrite (NO2 -) released by mTAL cells was determined using the modified Griess reagent method and taken as an index of nitric oxide production. The cultured cells were treated with various concentrations of CsA and different signal transduction regulators to assess the effect and possible pathway(s) of action of CsA on NO production in mTAL cells. Results. The basal production of NO by mTAL cells increased by 1.8-fold following incubation with bacterial lipopolysaccaride (LPS). Both aminoguanidine and L-NAME inhibited NO production. CsA (10-300 ng/ml) also inhibited NO production in a dose-dependent manner and prevented its increase induced by LPS. Phorbol 12-myristate 13-acetate (PMA), a PKC stimulator, enhanced slightly the production of NO under basal conditions and prevented the inhibitory action of CsA on NO production. These results suggest that the NO secreted by mouse cultured mTAL cells is dependent on the PKC pathway. Conclusion. These results show that CsA may downregulate the production of NO by cultured mTAL cells expressing iNOS mRNA and that the PKC pathway is involved in this process.",
keywords = "Cultured cells, Cyclosporin, Kidney, Nephrotoxicity, Nitric oxide, Thick ascending limbs",
author = "Wu, {Mai Szu} and Yang, {Chih Wei} and Marcelle Bens and Yu, {Hsiao Mei} and Huang, {Jeng Yi} and Wu, {Chin Herng} and Huang, {Chiu Ching} and Alain Vandewalle",
year = "1998",
month = "11",
language = "English",
volume = "13",
pages = "2814--2820",
journal = "Nephrology Dialysis Transplantation",
issn = "0931-0509",
publisher = "Oxford University Press",
number = "11",

}

TY - JOUR

T1 - Cyclosporin inhibits nitric oxide production in medullary ascending limb cultured cells

AU - Wu, Mai Szu

AU - Yang, Chih Wei

AU - Bens, Marcelle

AU - Yu, Hsiao Mei

AU - Huang, Jeng Yi

AU - Wu, Chin Herng

AU - Huang, Chiu Ching

AU - Vandewalle, Alain

PY - 1998/11

Y1 - 1998/11

N2 - Background. Nitric oxide (NO) has been shown to play a role in cyclosporin (CsA) nephrotoxicity, but its mechanism of action is still unclear. As inducible NO synthase (iNOS) mRNA has been found to be expressed in rat medullary thick ascending limb (mTAL) cells, we investigated the effects of CsA on NO production in a model of mouse cultured mTAL cells. Materials and methods. The experiments were carried out on sub-cultured cells derived from isolated mTAL microdissected from the kidney of C57BL/6 mice. The identification of the iNOS mRNA in mTAL microdissected segment and cultured cell was confirmed by RT-PCR and RsaI digestion. Nitrite (NO2 -) released by mTAL cells was determined using the modified Griess reagent method and taken as an index of nitric oxide production. The cultured cells were treated with various concentrations of CsA and different signal transduction regulators to assess the effect and possible pathway(s) of action of CsA on NO production in mTAL cells. Results. The basal production of NO by mTAL cells increased by 1.8-fold following incubation with bacterial lipopolysaccaride (LPS). Both aminoguanidine and L-NAME inhibited NO production. CsA (10-300 ng/ml) also inhibited NO production in a dose-dependent manner and prevented its increase induced by LPS. Phorbol 12-myristate 13-acetate (PMA), a PKC stimulator, enhanced slightly the production of NO under basal conditions and prevented the inhibitory action of CsA on NO production. These results suggest that the NO secreted by mouse cultured mTAL cells is dependent on the PKC pathway. Conclusion. These results show that CsA may downregulate the production of NO by cultured mTAL cells expressing iNOS mRNA and that the PKC pathway is involved in this process.

AB - Background. Nitric oxide (NO) has been shown to play a role in cyclosporin (CsA) nephrotoxicity, but its mechanism of action is still unclear. As inducible NO synthase (iNOS) mRNA has been found to be expressed in rat medullary thick ascending limb (mTAL) cells, we investigated the effects of CsA on NO production in a model of mouse cultured mTAL cells. Materials and methods. The experiments were carried out on sub-cultured cells derived from isolated mTAL microdissected from the kidney of C57BL/6 mice. The identification of the iNOS mRNA in mTAL microdissected segment and cultured cell was confirmed by RT-PCR and RsaI digestion. Nitrite (NO2 -) released by mTAL cells was determined using the modified Griess reagent method and taken as an index of nitric oxide production. The cultured cells were treated with various concentrations of CsA and different signal transduction regulators to assess the effect and possible pathway(s) of action of CsA on NO production in mTAL cells. Results. The basal production of NO by mTAL cells increased by 1.8-fold following incubation with bacterial lipopolysaccaride (LPS). Both aminoguanidine and L-NAME inhibited NO production. CsA (10-300 ng/ml) also inhibited NO production in a dose-dependent manner and prevented its increase induced by LPS. Phorbol 12-myristate 13-acetate (PMA), a PKC stimulator, enhanced slightly the production of NO under basal conditions and prevented the inhibitory action of CsA on NO production. These results suggest that the NO secreted by mouse cultured mTAL cells is dependent on the PKC pathway. Conclusion. These results show that CsA may downregulate the production of NO by cultured mTAL cells expressing iNOS mRNA and that the PKC pathway is involved in this process.

KW - Cultured cells

KW - Cyclosporin

KW - Kidney

KW - Nephrotoxicity

KW - Nitric oxide

KW - Thick ascending limbs

UR - http://www.scopus.com/inward/record.url?scp=0031756772&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0031756772&partnerID=8YFLogxK

M3 - Article

C2 - 9829483

AN - SCOPUS:0031756772

VL - 13

SP - 2814

EP - 2820

JO - Nephrology Dialysis Transplantation

JF - Nephrology Dialysis Transplantation

SN - 0931-0509

IS - 11

ER -