Ciclosporin reduces paracellin-1 expression and magnesium transport in thick ascending limb cells

Chiz Tzung Chang, Cheng Chieh Hung, Ya Chung Tian, Chih Wei Yang, Mai Szu Wu

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Background. Renal magnesium (Mg 2+) wasting is one of the ciclosporin (CsA) tubular effects. The major site of Mg 2+ transport is the thick ascending limb (TAL), where 70% of the ultrafiltrable Mg 2+ is reabsorbed paracellularly. Paracellin-1 is a tight junction protein, which regulates the paracellular Mg 2+ transport in the TAL. We hypothesize that CsA reduces the expression and function of paracellin-1 and accounts for the observed renal Mg 2+ wasting. Methods. We established an immortalized cultured cortical TAL (cTAL) cell line from L-PK/Tag1 transgenic mice by microdissection. The cultured cells expressed paracellin-1 and the characteristics of cTAL cells. Real-time PCR and western blotting were used to test the CsA effects on paracellin-1 expression of cultured cTAL cells. Cytosolic-free Mg 2+ concentration [Mg 2+] i change with time in a single cTAL cell was used as an indicator of transcellular Mg 2+ transport and assessed by using fluorescence dye Mag-fura-2 AM. Paracellular Mg 2+ transport was measured by cells grown in porous filters. Results. The results showed that CsA significantly reduced paracellin-1 mRNA and protein expression in a dose-dependent manner. CsA (100 ng/ml) incubation for 24 h induced a decrease of paracellin-1 mRNA by 89.4% and paracellin-1 protein by 75.4%. CsA (100 ng/ml) did not change transcellular Mg 2+ transport, but paracellular Mg 2+ transport was decreased in CsA-treated cTAL cells by 74.4%. Conclusion. These results suggested that reduced PCLN-1 expression and paracellular Mg 2+ transport might play a role in the renal Mg 2+ wasting in the CsA tubular effect.

Original languageEnglish
Pages (from-to)1033-1040
Number of pages8
JournalNephrology Dialysis Transplantation
Volume22
Issue number4
DOIs
Publication statusPublished - Apr 1 2007
Externally publishedYes

Fingerprint

Magnesium
Cyclosporine
Extremities
Kidney
Tight Junction Proteins
Microdissection
Messenger RNA
Transgenic Mice
Real-Time Polymerase Chain Reaction
Cultured Cells
Proteins
Coloring Agents
Fluorescence
Western Blotting
Cell Line

Keywords

  • Ciclosporin
  • Magnesium transport
  • Paracellin-1
  • PCLN-1
  • Thick ascending limb

ASJC Scopus subject areas

  • Nephrology
  • Transplantation

Cite this

Ciclosporin reduces paracellin-1 expression and magnesium transport in thick ascending limb cells. / Chang, Chiz Tzung; Hung, Cheng Chieh; Tian, Ya Chung; Yang, Chih Wei; Wu, Mai Szu.

In: Nephrology Dialysis Transplantation, Vol. 22, No. 4, 01.04.2007, p. 1033-1040.

Research output: Contribution to journalArticle

Chang, Chiz Tzung ; Hung, Cheng Chieh ; Tian, Ya Chung ; Yang, Chih Wei ; Wu, Mai Szu. / Ciclosporin reduces paracellin-1 expression and magnesium transport in thick ascending limb cells. In: Nephrology Dialysis Transplantation. 2007 ; Vol. 22, No. 4. pp. 1033-1040.
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abstract = "Background. Renal magnesium (Mg 2+) wasting is one of the ciclosporin (CsA) tubular effects. The major site of Mg 2+ transport is the thick ascending limb (TAL), where 70{\%} of the ultrafiltrable Mg 2+ is reabsorbed paracellularly. Paracellin-1 is a tight junction protein, which regulates the paracellular Mg 2+ transport in the TAL. We hypothesize that CsA reduces the expression and function of paracellin-1 and accounts for the observed renal Mg 2+ wasting. Methods. We established an immortalized cultured cortical TAL (cTAL) cell line from L-PK/Tag1 transgenic mice by microdissection. The cultured cells expressed paracellin-1 and the characteristics of cTAL cells. Real-time PCR and western blotting were used to test the CsA effects on paracellin-1 expression of cultured cTAL cells. Cytosolic-free Mg 2+ concentration [Mg 2+] i change with time in a single cTAL cell was used as an indicator of transcellular Mg 2+ transport and assessed by using fluorescence dye Mag-fura-2 AM. Paracellular Mg 2+ transport was measured by cells grown in porous filters. Results. The results showed that CsA significantly reduced paracellin-1 mRNA and protein expression in a dose-dependent manner. CsA (100 ng/ml) incubation for 24 h induced a decrease of paracellin-1 mRNA by 89.4{\%} and paracellin-1 protein by 75.4{\%}. CsA (100 ng/ml) did not change transcellular Mg 2+ transport, but paracellular Mg 2+ transport was decreased in CsA-treated cTAL cells by 74.4{\%}. Conclusion. These results suggested that reduced PCLN-1 expression and paracellular Mg 2+ transport might play a role in the renal Mg 2+ wasting in the CsA tubular effect.",
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T1 - Ciclosporin reduces paracellin-1 expression and magnesium transport in thick ascending limb cells

AU - Chang, Chiz Tzung

AU - Hung, Cheng Chieh

AU - Tian, Ya Chung

AU - Yang, Chih Wei

AU - Wu, Mai Szu

PY - 2007/4/1

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N2 - Background. Renal magnesium (Mg 2+) wasting is one of the ciclosporin (CsA) tubular effects. The major site of Mg 2+ transport is the thick ascending limb (TAL), where 70% of the ultrafiltrable Mg 2+ is reabsorbed paracellularly. Paracellin-1 is a tight junction protein, which regulates the paracellular Mg 2+ transport in the TAL. We hypothesize that CsA reduces the expression and function of paracellin-1 and accounts for the observed renal Mg 2+ wasting. Methods. We established an immortalized cultured cortical TAL (cTAL) cell line from L-PK/Tag1 transgenic mice by microdissection. The cultured cells expressed paracellin-1 and the characteristics of cTAL cells. Real-time PCR and western blotting were used to test the CsA effects on paracellin-1 expression of cultured cTAL cells. Cytosolic-free Mg 2+ concentration [Mg 2+] i change with time in a single cTAL cell was used as an indicator of transcellular Mg 2+ transport and assessed by using fluorescence dye Mag-fura-2 AM. Paracellular Mg 2+ transport was measured by cells grown in porous filters. Results. The results showed that CsA significantly reduced paracellin-1 mRNA and protein expression in a dose-dependent manner. CsA (100 ng/ml) incubation for 24 h induced a decrease of paracellin-1 mRNA by 89.4% and paracellin-1 protein by 75.4%. CsA (100 ng/ml) did not change transcellular Mg 2+ transport, but paracellular Mg 2+ transport was decreased in CsA-treated cTAL cells by 74.4%. Conclusion. These results suggested that reduced PCLN-1 expression and paracellular Mg 2+ transport might play a role in the renal Mg 2+ wasting in the CsA tubular effect.

AB - Background. Renal magnesium (Mg 2+) wasting is one of the ciclosporin (CsA) tubular effects. The major site of Mg 2+ transport is the thick ascending limb (TAL), where 70% of the ultrafiltrable Mg 2+ is reabsorbed paracellularly. Paracellin-1 is a tight junction protein, which regulates the paracellular Mg 2+ transport in the TAL. We hypothesize that CsA reduces the expression and function of paracellin-1 and accounts for the observed renal Mg 2+ wasting. Methods. We established an immortalized cultured cortical TAL (cTAL) cell line from L-PK/Tag1 transgenic mice by microdissection. The cultured cells expressed paracellin-1 and the characteristics of cTAL cells. Real-time PCR and western blotting were used to test the CsA effects on paracellin-1 expression of cultured cTAL cells. Cytosolic-free Mg 2+ concentration [Mg 2+] i change with time in a single cTAL cell was used as an indicator of transcellular Mg 2+ transport and assessed by using fluorescence dye Mag-fura-2 AM. Paracellular Mg 2+ transport was measured by cells grown in porous filters. Results. The results showed that CsA significantly reduced paracellin-1 mRNA and protein expression in a dose-dependent manner. CsA (100 ng/ml) incubation for 24 h induced a decrease of paracellin-1 mRNA by 89.4% and paracellin-1 protein by 75.4%. CsA (100 ng/ml) did not change transcellular Mg 2+ transport, but paracellular Mg 2+ transport was decreased in CsA-treated cTAL cells by 74.4%. Conclusion. These results suggested that reduced PCLN-1 expression and paracellular Mg 2+ transport might play a role in the renal Mg 2+ wasting in the CsA tubular effect.

KW - Ciclosporin

KW - Magnesium transport

KW - Paracellin-1

KW - PCLN-1

KW - Thick ascending limb

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