Journal List > Korean J Physiol Pharmacol > v.15(5) > 1025750

Bae, Kim, Ma, Lee, and Kim: Altered Regulation of Renal Nitric Oxide and Atrial Natriuretic Peptide Systems in Lipopolysaccharide-induced Kidney Injury


Nitric oxide (NO) and atrial natriuretic peptide (ANP) may induce vascular relaxation by increasing the production of cyclic guanosine monophosphate (cGMP), an important mediator of vascular tone during sepsis. This study aimed to determine whether regulation of NO and the ANP system is altered in lipopolysaccharide (LPS)-induced kidney injury. LPS (10 mg · kg–1) was injected in the tail veins of male Sprague-Dawley rats; 12 hours later, the kidneys were removed. Protein expression of NO synthase (NOS) and neutral endopeptidase (NEP) was determined by semiquantitative immunoblotting. As an index of synthesis of NO, its stable metabolites (nitrite/nitrate, NOx) were measured using colorimetric assays. mRNA expression of the ANP system was determined by real-time polymerase chain reaction. To determine the activity of guanylyl cyclase (GC), the amount of cGMP generated in response to sodium nitroprusside (SNP) and ANP was calculated. Creatinine clearance decreased and fractional excretion of sodium increased in LPS-treated rats compared with the controls. Inducible NOS protein expression increased in LPS-treated rats, while that of endothelial NOS, neuronal NOS, and NEP remained unchanged. Additionally, urinary and plasma NOx levels increased in LPS-treated rats. SNP-stimulated GC activity remained unchanged in the glomerulus and papilla in the LPS-treated rats. mRNA expression of natriuretic peptide receptor (NPR)-C decreased in LPS-treated rats, while that of ANP and NPR-A did not change. ANP-stimulated GC activity reduced in the glomerulus and papilla. In conclusion, enhancement of the NO/cGMP pathway and decrease in ANP clearance were found play a role in the pathogenesis of LPS-induced kidney injury.


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Fig. 1.
Semiquantitative immunoblotting of endothelial nitric oxide synthase (eNOS), inducible NOS (iNOS) and neuronal NOS (nNOS) in the kidney. p<0.05 compared with control.
Fig. 2.
Nitric oxide metabolites (nitrite/nitrate, NOx) in plasma and urine. p<0.05 compared with control.
Fig. 3.
(A) Semiquantitative immunoblotting of soluble guanylnyl cyclase (sGC) in the kidney. (B) cGMP production in response to sodium nitroprusside (SNP) in the glomerulus and papilla. Each point represents mean±SEM of experimental rats. p<0.05 compared with control.
Fig. 4.
The mRNA expression of atrial natriuretic peptide (ANP), natriuretic peptide receptor type A and C (NPR-A, NPR-C) in whole kidney. Columns show real-time PCR data representing control and LPS groups. p<0.05 compared with control.
Fig. 5.
(A) Semiquantitative immunoblotting of neutral endopeptidase (NEP) in the kidney. (B) cGMP production in response to atrial natriuretic peptide (ANP) in the glomerulus and papilla. Each point represents mean±SEM of experimental rats. p<0.05 compared with control.
Table 1.
Changes in blood pressure and renal functional data
  Control (n=8) LPS (n=9)
Body weight (g) 200±5.0 200±6.3
UO (ml/12 hr) 9.9±2.6 8.4±2.4
P-Cr (mg/dl) 0.25±0.07 0.73±015
Ccr (ml/min) 1.16±0.34 0.26±0.14
FENa (%) 0.47±0.18 1.36±0.19

Values are expressed as mean±SEM. These values are measured on the last day of experiments. LPS, lipopolysaccharide induced endotoxemia rat model; UO, urine output; P-Cr, plasma creatinine; Ccr, creatinine clearance; FENa, fractional excretion of sodium into urine.

p<0.05 compared with control.

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