Yonsei Med J.  2002 Apr;43(2):183-192. 10.3349/ymj.2002.43.2.183.

Inducible Nitric Oxide Synthase (iNOS) Expression Is Increased in Lipopolysaccharide (LPS)-Stimulated Diabetic Rat Glomeruli: Effect of ACE Inhibitor and Angiotensin II Receptor Blocker

• Lee HY
• Noh HJ
• Gang JG
• Xu ZG
• Jeong HJ
• Kang SW
• Choi KH
• Han DS

• KMID: 1118967
• DOI: http://doi.org/10.3349/ymj.2002.43.2.183

Abstract

Previously, we reported that high glucose enhanced cytokine-induced nitric oxide (NO) production by rat mesangial cells (MCs), and that the enhanced expression of the iNOS pathway may promote extracellular matrix accumulation by MCs. The present study was designed to examine whether the iNOS pathway is pathologically altered in experimental diabetic nephropathy, and whether therapy with angiotensin converting enzyme (ACE) inhibitor (imidapril: I) or angiotensin II type I receptor (AT1) blocker (L-158,809: L), ameliorates these changes. Male Sprague-Dawley rats were injected with diluent (control: C) or streptozotocin. At sacrifice after 4, 8 and 12 weeks, rats underwent either a 4 hour placebo or an intraperitoneal lipopolysaccharide (LPS, 2 mg/kg) challenge. Systolic blood pressure (SBP) and urinary protein excretion (UPE) increased significantly in diabetic (D) rats compared with C. The basal expression of glomerular iNOS mRNA was increased in D rats compared with that of C rats, by reverse- transcription (RT)-polymerase chain reaction (PCR), whereas there was no significant difference in the level of protein by Western blot analysis. Upon LPS stimulation, the iNOS mRNA and protein expression was significantly elevated in D rats. In D rats, this up-regulation, of LPS-stimulated iNOS expression, was equally ameliorated both by I and L in mRNA and protein levels. From immunohistochemistry (IHC), there was a negative staining for the iNOS within the glomeruli of five C rats without LPS treatment, but one of four rats, with LPS treatment, showed minimal iNOS staining in the glomeruli. In D rats, the glomerular mesangium and podocytes were positive for iNOS in each of three out of five rats with, and without, LPS treatment. In conclusion, LPS-stimulated glomerular iNOS expression was enhanced in diabetic pnephropathy, and the activation of angiotensin II may play a role in this enhancement.

Keyword

iNOS; diabetic nephropathy; angiotensin II

MeSH Terms

Angiotensin-Converting Enzyme Inhibitors/pharmacology
Animal
Kidney Glomerulus/*drug effects/*enzymology
Lipopolysaccharides/*pharmacology
Male
Nitric-Oxide Synthase/*metabolism
Rats
Rats, Sprague-Dawley
Receptors, Angiotensin/antagonists & inhibitors