Korean J Physiol Pharmacol.  2011 Oct;15(5):273-277. 10.4196/kjpp.2011.15.5.273.

Altered Regulation of Renal Nitric Oxide and Atrial Natriuretic Peptide Systems in Lipopolysaccharide-induced Kidney Injury

• Bae EH
• Kim IJ
• Ma SK
• Lee JU
• Kim SW

• KMID: 2071729
• DOI: http://doi.org/10.4196/kjpp.2011.15.5.273

Abstract

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.

Keyword

Lipopolysaccharide; Atrial natriuretic peptide; Nitric oxide; Guanylyl cyclase

MeSH Terms

Animals
Atrial Natriuretic Factor
Creatinine
Guanosine Monophosphate
Guanylate Cyclase
Humans
Immunoblotting
Kidney
Male
Neprilysin
Neurons
Nitric Oxide
Nitric Oxide Synthase
Nitric Oxide Synthase Type II
Nitroprusside
Plasma
Rats
Real-Time Polymerase Chain Reaction
Receptors, Peptide
Relaxation
RNA, Messenger
Sepsis
Sodium
Veins
Atrial Natriuretic Factor
Creatinine
Guanosine Monophosphate
Guanylate Cyclase
Neprilysin
Nitric Oxide
Nitric Oxide Synthase
Nitric Oxide Synthase Type II
Nitroprusside
RNA, Messenger
Receptors, Peptide
Sodium

Figure

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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.

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  Fig. 2. Nitric oxide metabolites (nitrite/nitrate, NOx) in plasma and urine. ∗p<0.05 compared with control.

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  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.

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  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.

Reference

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