J Korean Med Sci.  2010 Sep;25(9):1305-1312. 10.3346/jkms.2010.25.9.1305.

High Dose Vitamin D3 Attenuates the Hypocalciuric Effect of Thiazide in Hypercalciuric Rats

• Jang HR
• Lee JW
• Kim S
• Heo NJ
• Lee JH
• Kim HS
• Jung JY
• Oh YK
• Na KY
• Han JS
• Joo KW

• KMID: 1785909
• DOI: http://doi.org/10.3346/jkms.2010.25.9.1305

Abstract

Thiazide is known to decrease urinary calcium excretion. We hypothesized that thiazide shows different hypocalciuric effects depending on the stimuli causing hypercalciuria. The hypocalciuric effect of hydrochlorothiazide (HCTZ) and the expression of transient receptor potential vanilloid 5 (TRPV5), calbindin-D(28K), and several sodium transporters were assessed in hypercalciuric rats induced by high calcium diet and vitamin D3. Urine calcium excretion and the expression of transporters were measured from 4 groups of Sprague-Dawley rats; control, HCTZ, high calcium-vitamin D, and high calcium-vitamin D with HCTZ groups. HCTZ decreased urinary calcium excretion by 51.4% in the HCTZ group and only 15% in the high calcium-vitamin D with HCTZ group. TRPV5 protein abundance was not changed by HCTZ in the high calcium-vitamin D with HCTZ group compared to the high calcium-vitamin D group. Protein abundance of NHE3, SGLT1, and NKCC2 decreased in the hypercalciuric rats, and only SGLT1 protein abundance was increased by HCTZ in the hypercalciuric rats. The hypocalciuric effect of HCTZ is attenuated in high calcium and vitamin D-induced hypercalciuric rats. This attenuation seems to have resulted from the lack of HCTZ's effect on protein abundance of TRPV5 in severe hypercalciuric condition induced by high calcium and vitamin D.

Keyword

Hypercalciuria; TRPV5; Sodium Transporters; Cholecalciferol; Thiazides

MeSH Terms

Animals
Calcium/therapeutic use/urine
Calcium Channels/genetics/metabolism
Cholecalciferol/*toxicity
Hydrochlorothiazide/*therapeutic use
Hypercalciuria/chemically induced/*drug therapy
Rats
Rats, Sprague-Dawley
Sodium Chloride Symporter Inhibitors/*therapeutic use
Sodium-Glucose Transporter 1/genetics/metabolism
Sodium-Hydrogen Antiporter/genetics/metabolism
Sodium-Potassium-Chloride Symporters/genetics/metabolism
TRPV Cation Channels/genetics/metabolism

Figure

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  Fig. 1 Serial follow up of urine Ca/Cr ratio in high calcium diet and/or vitamin D-induced hypercalciuric rats. Calcitriol was injected into the peritoneal cavities of rats in the vitamin D group and the high Ca with vitamin D group on day 4. The urine Ca/Cr ratio of the high Ca with vitamin D group was highest on day 5. *P=0.01 when compared to the control group; †P=0.01 when compared to the high Ca group; ‡P=0.01 when compared to the vitamin D group. High Ca, high calcium diet group; vitamin D, vitamin D group; high Ca & vitamin D, high calcium diet with vitamin D group.

• 

  Fig. 2 Urine Ca/Cr ratio on day 6 in high calcium diet and vitamin D-induced hypercalciuric rats. Urine Ca/Cr was significantly decreased in the HCTZ group and increased in the high Ca-vitamin D group. In the high Ca-vitamin D with HCTZ group, urine Ca/Cr was decreased to about 85% of the high Ca-vitamin D group. *P=0.01 when compared to the control group. HCTZ, HCTZ (hydrochlorothiazide) group; Ca-VitD, high calcium diet and vitamin D group; Ca-VitD with HCTZ, high calcium diet and vitamin D with HCTZ group.

• 

  Fig. 3 TRPV5 expression in high calcium diet and vitamin D-induced hypercalciuric rats. (A) TRPV5 mRNA expression was significantly decreased in the high Ca-vitamin D group and increased by HCTZ administration. (B) TRPV5 protein abundance was significantly increased in the HCTZ group, but decreased in both high Ca-vitamin D and high Ca-vitamin D with HCTZ groups. *P=0.02 when compared to the control group; †P=0.02 when compared to the high Ca-vitamin D group. HCTZ, HCTZ (hydrochlorothiazide) group; Ca-VitD, high calcium diet and vitamin D group; Ca-VitD with HCTZ, high calcium diet and vitamin D with HCTZ group.

• 

  Fig. 4 Calbindin-D28K expression in high calcium diet and vitamin D-induced hypercalciuric rats. (A) Expression of calbindin-D28K mRNA was significantly decreased in the high Ca-vitamin D group and increased when HCTZ was administered. (B) Protein abundance of calbindin-D28K was not different among groups. *P=0.01 when compared to the control group; †P=0.02 when compared to the high Ca-vitamin D group. HCTZ, HCTZ (hydrochlorothiazide) group; Ca-VitD, high calcium diet and vitamin D group; Ca-VitD with HCTZ, high calcium diet and vitamin D with HCTZ group.

• 

  Fig. 5 Protein abundance of sodium transporters in high calcium diet and vitamin D-induced hypercalciuric rats. Protein abundance of NHE3 (A), SGLT1 (B), NKCC2 (C), and NCC (D) was decreased in the high Ca-vitamin D group. SGLT1 protein abundance was increased in the high Ca-vitamin D with HCTZ group compared to the high Ca-vitamin D group, but protein abundance of NHE3, NKCC2, and NCC was not changed by HCTZ. *P=0.01 when compared to the control group; †P=0.02 when compared to the high Ca-vitamin D group. HCTZ, HCTZ (hydrochlorothiazide) group; Ca-VitD, high calcium diet and vitamin D group; Ca-VitD with HCTZ, high calcium diet and vitamin D with HCTZ group.

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