Altered Renal Expression of Acid-base Transporters in Rats with Glycerol-induced Tubular Injury

Seong Kwon Ma; Eun Hui Bae; JongUn Lee; Soo Wan Kim

doi:10.4068/cmj.2010.46.3.163

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Ma, Bae, Lee, and Kim: Altered Renal Expression of Acid-base Transporters in Rats with Glycerol-induced Tubular Injury

Original Article

Chonnam Medical Journal

Chonnam Medical Journal 2010; 46(3): 163-169.

Published online: 23 December 2010

DOI: https://doi.org/10.4068/cmj.2010.46.3.163

Altered Renal Expression of Acid-base Transporters in Rats with Glycerol-induced Tubular Injury

Seong Kwon Ma, Eun Hui Bae, JongUn Lee¹, Soo Wan Kim

Department of Internal Medicine, Chonnam National University Medical School, Gwangju, Korea.

¹Department of Physiology, Chonnam National University Medical School, Gwangju, Korea.

Corresponding author: skimw@chonnam.ac.kr

Received 8 November 2010 Accepted 16 November 2010

Abstract

The present study aimed to investigate the altered regulation of renal tubular acid-base transporters in rats with glycerol-induced tubular injury. Male Sprague-Dawley rats were used. Rats were injected with 50% glycerol in normal saline (7 mL/kg, i.m.) after water deprivation for 12 hours and were then sacrificed at 24 hours after the glycerol injection. The expression of Na,K-ATPase α1-subunit, type 3 Na⁺/H⁺ exchanger (NHE3), type 1 Na⁺:HCO₃^- (NBC1), and B1-subunit of apical H⁺-ATPase was determined in the kidney by semiquantitative immunoblotting and immunohistochemistry. In the experimental rats, creatinine clearance was decreased, whereas fractional sodium excretion was not changed. Urine pH and bicarbonate concentrations were decreased, although plasma pH and bicarbonate concentrations were not changed. In the experimental group, the protein expression of Na,K-ATPase α1-subunit was decreased in the cortex and outer stripe of outer medulla (cortex/OSOM) and inner stripe of outer medulla (ISOM) but increased in the inner medulla. The expression of NHE3 was decreased in the cortex/OSOM and ISOM, and that of NBC1 was decreased in the cortex/OSOM. By contrast, the expression of H⁺-ATPase was increased in the cortex/OSOM and inner medulla but was unchanged in the ISOM. Immunohistochemical analyses confirmed the immunoblotting data. In glycerol-induced tubular injury, the down-regulation of Na,K-ATPase α1-subunit, NHE3, and NBC1 may contribute to impaired tubular reabsorption of sodium and bicarbonate. The upregulation of H⁺-ATPase may play a preventive role against the development of metabolic acidosis.

Keywords: Sodium-hydrogen exchanger 3, Sodium, Bicarbonates, Membrain transport proteins

Figures and Tables

Fig. 1

(A) Semiquantitative immunoblotting of Na,K-ATPase α1-subunit in the cortex and outer stripe of outer medulla (cortex/OSOM) of the kidney. Exp, experimental. Each column represents mean±SEM of 8 rats. ^*p<0.05 compared with control kidney. (B) Immunoperoxidase microscopy of Na,K-ATPase α1-subunit in the cortex of the kidney. PT, proximal tubule.

Fig. 2

(A) Semiquantitative immunoblotting of type 3 Na⁺/H⁺ exchanger (NHE3) in the cortex/OSOM of the kidney. (B) Immunoperoxidase microscopy of NHE3 in the cortex of the kidney. Legends as in Figure 1.

Fig. 3

(A) Semiquantitative immunoblotting of type 1 Na⁺:HCO₃^- cotransporter (NBC1) in the cortex/OSOM of the kidney. (B) Immunoperoxidase microscopy of NBC1 in the cortex of the kidney. Legends as in Figure 1.

Fig. 4

(A) Semiquantitative immunoblotting of H⁺-ATPase (B1-subunit) in the cortex/OSOM of the kidney. (B) Immunoperoxidase microscopy of H⁺-ATPase (B1-subunit) in the kidney. CCD, cortical collecting duct. Legends as in Figure 1.

Table 1

Parameters of renal function and acid-base status

Values are mean±SEM. ^*p<0.05 compared with control rats. Exp, experimental; Cr, creatinine; Ccr, creatinine clearance; FE_Na, fractional excretion of sodium into urine.

Table 2

The protein expression of Na,K-ATPase and acid-base transporters

Values are mean±SEM. Exp, experimental; OSOM, outer stripe of outer medulla; ISOM, inner stripe of outer medulla; NHE3, type 3 Na⁺/H⁺ exchanger; NBC1, type 1 Na⁺:HCO₃^- cotransporter. ^*p<0.05 compared with control rats.

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