J Korean Med Sci.  2009 Jan;24(Suppl 1):S161-S169. 10.3346/jkms.2009.24.S1.S161.

Effects of Thiazide on the Expression of TRPV5, Calbindin-D28K, and Sodium Transporters in Hypercalciuric Rats

• Jang HR
• Kim S
• Heo NJ
• Lee JH
• Kim HS
• Nielsen S
• Jeon US
• Oh YK
• Na KY
• Joo KW
• Han JS

• KMID: 1778157
• DOI: http://doi.org/10.3346/jkms.2009.24.S1.S161

Abstract

TRPV5 is believed to play an important role in the regulation of urinary calcium excretion. We assessed the effects of hydrochlorothiazide (HCTZ) on the expression of TRPV5, calbindin-D28K, and several sodium transporters in hypercalciuric rats. Sprague- Dawley rats were divided into 4 groups; control, HCTZ, high salt, and high salt with HCTZ group in experiment 1; control, HCTZ, high calcium (Ca), and high Ca with HCTZ group in experiment 2. To quantitate the expression of TRPV5, calbindin- D28K, and sodium transporters, western blotting was performed. In both experiments, HCTZ significantly decreased urinary calcium excretion. TRPV5 protein abundance decreased in all hypercalciuric rats, and restored by HCTZ in both high salt with HCTZ and high Ca with HCTZ group. Calbindin-D28K protein abundance increased in the high salt and high salt with HCTZ groups, but did not differ among groups in experiment 2. Protein abundance of NHE3 and NKCC2 decreased in all hypercalciuric rats, and were restored by HCTZ in only high Ca-induced hypercalciuric rats. In summary, protein abundance of TRPV5, NHE3, and NKCC2 decreased in all hypercalciuric rats. The hypocalciuric effect of HCTZ is associated with increased protein abundance of TRPV5 in high salt or calcium diet-induced hypercalciuric rats.

Keyword

Hypercalciuria; Thiazides; Transporters

MeSH Terms

Animals
Biological Transport
Calcium/urine
Calcium Channels/chemistry
Calcium-Binding Protein, Vitamin D-Dependent/*biosynthesis
Hydrochlorothiazide/pharmacology
Hypercalciuria/*therapy
Male
Models, Biological
Rats
Rats, Sprague-Dawley
Sodium/*metabolism
Sodium-Hydrogen Antiporter/chemistry
Sodium-Potassium-Chloride Symporters/metabolism
TRPV Cation Channels/*biosynthesis/chemistry
Thiazides/*pharmacology

Figure

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  Fig. 1 Urine Ca/Cr ratio on day 7 in high salt diet-induced hypercalciuric rats. Urine Ca/Cr was significantly increased in the high salt group, and decreased in the high salt with HCTZ group when compared to the high salt group. *p<0.05 when compared to the control group; †p<0.05 when compared to the high salt group.

• 

  Fig. 2 TRPV5 expression in high salt diet-induced hypercalciuric rats. TRPV5 protein abundance was significantly increased in the HCTZ group, decreased in the high salt group, and increased in the high salt with HCTZ group when compared to the high salt group. *p<0.05 when compared to the control group; †p<0.05 when compared to the high salt group.

• 

  Fig. 3 Calbindin-D28K expression in high salt diet-induced hypercalciuric rats. Calbindin-D28K protein abundance was significantly increased in both high salt and high salt with HCTZ groups. *p<0.05 when compared to the control group.

• 

  Fig. 4 Protein abundance of sodium transporters in high salt diet-induced hypercalciuric rats. Protein abundance of NHE3 and NKCC2 was increased in the HCTZ group, but decreased in the high salt group. NCC protein abundance increased in the HCTZ group, but was not changed in the high salt group. SGLT1 protein abundance increased in both HCTZ group and high salt group. *p<0.05 when compared to the control group.

• 

  Fig. 5 Urine Ca/Cr ratio on day 7 in high calcium diet-induced hypercalciuric rats. Urine Ca/Cr was significantly increased in the high Ca group. In the high Ca with HCTZ group, urine Ca/Cr was decreased to about 47% of the high Ca group. *p<0.05 when compared to the control group; †p<0.05 when compared to the high Ca group.

• 

  Fig. 6 TRPV5 expression in high calcium diet-induced hypercalciuric rats. TRPV5 protein abundance was significantly decreased in the high Ca group, but restored in the high Ca with HCTZ group. *p<0.05 when compared to the control group; †p<0.05 when compared to the high Ca group.

• 

  Fig. 7 Calbindin-D28K expression in high calcium diet-induced hypercalciuric rats. Protein abundance of calbindin-D28K was not different among groups.

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  Fig. 8 Protein abundance of sodium transporters in high calcium diet-induced hypercalciuric rats. Protein abundance of NHE3, SGLT1, NKCC2, and NCC was decreased in the high Ca group. Protein abundance of NHE3, SGLT1, and NKCC2 was increased by HCTZ in the high Ca with HCTZ group when compared to the high Ca group. *p<0.05 when compared to the control group; †p<0.05 when compared to the high Ca group.

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