Chronic kidney disease is accompanied by nitric oxide (NO) deficiency and oxidative stress, which contribute to progression. We investigated whether the antioxidant vitamin E could preserve renal function and NO bioavailability and reduce oxidative stress in the 5/6th nephrectomy (NX) rat model. We studied the following three groups of male Sprague-Dawley rats: sham (n = 6), 5/6 NX control (n = 6), and 5/6 NX treated with vitamin E (5,000 IU/kg chow; n = 5). The 5/6 NX group showed increased severity of glomerulosclerosis vs. sham, and this was ameliorated by vitamin E therapy. Both 5/6 NX groups showed similar elevations in plasma creatinine and proteinuria and decreased 24-h creatinine clearance compared with sham. There was increased NADPH-dependent superoxide production in 5/6 NX rats vs. sham that was prevented by vitamin E. Total NO production was similarly reduced in both 5/6 NX groups. There was unchanged abundance of endothelial nitric oxide synthesis (NOS) in renal cortex and medulla and neuronal (n) NOS in medulla. However, in kidney cortex, 5/6 NX rats had lower nNOS abundance than sham, which was restored by vitamin E. An increased plasma asymmetric dimethylarginine occurred with 5/6 NX associated with decreased renal dimethylarginine dimethylaminohydrolase activity and increased type 1 protein arginine methyltransferase expression.

The inbred obese Zucker (ZDF/Gmi, fa/fa) rat develops severe hyperglycemia and also exhibits severe renal disease. In this study, we compared the relative benefits of long-term treatment with angiotensin-converting enzyme inhibition (ACEI) to a peroxisome proliferator-activated receptor ␥ (PPAR␥) agonist. Four groups of obese inbred Zucker rats were studied over a 6-month observation period; untreated animals, rats treated with ACEI alone, rats treated with PPAR␥ agonist alone, and rats treated with a combination of ACEI and PPAR␥ agonist. PPAR␥ agonist treatment normalized plasma glucose and led to massive increases in body weight. Both ACEI and PPAR␥ agonist were effective in reducing the proteinuria and glomerular and tubular kidney damage. However, the PPAR␥ agonist exerted superior renal protection compared with ACEI, in this model of spontaneously occurring chronic renal disease in the diabetic, obese inbred Zucker rat. Of note, although ACEI lowered blood pressure, there was no difference in glomerular blood pressure in any group at the end of the study. The glomerular filtration rate (GFR) was improved by ACEI with a borderline effect of PPAR␥ agonist alone. A mild additive protection on GFR and tubulointerstitial damage was seen with the combination. Based on the literature it is likely that the superior protection by PPAR␥ agonist versus glomerular and tubular damage as well as proteinuria extends beyond glycemic and lipidmic control and also reflects direct, protective intrarenal actions of the PPAR␥ agonists.

Aims/hypothesis. In several other models of chronic renal disease, decreases in renal nitric oxide activity and nitric oxide synthase (NOS) protein abundance have been demonstrated. Here, we studied diabetic obese Zucker (ZDF Gmi fa/fa) rats that develop severe hyperglycaemia and renal disease, together with their lean control animals, to determine if renal nitric oxide deficiency also occurs in this model. Methods. Obese Zucker rats aged 10 to 12 weeks were maintained on Purina 5008 diet until 4, 8, or 11 months of age and compared with similarly maintained, 4-and 11-month-old lean Zucker rats. NOS activity and abundance of endothelial NOS (eNOS) and neuronal NOS (nNOS) were measured on homogenates of kidney cortex. Blood was analysed for glucose, lipids, creatinine, and blood urea nitrogen and kidney tissue was obtained for histology. Results. Obese rats exhibited severe hyperglycaemia from 4 months of age and developed increasing hyperlipidaemia, proteinuria, and decreasing renal function with age compared to lean counterparts. At 4 months cortical NOS activity and nNOS abundance were lower in obese rats than in lean ones. At 11 months NOS activity remained depressed and nNOS abundance had declined further in obese rats. Glomerulosclerosis in the obese rats was mild at 4 months, becoming severe by 11 months. Lean rats had only mild age-dependent increases in glomerular injury. Conclusions/interpretation. The chronic renal disease that occurs in hyperglycaemic, obese Zucker rats is associated with decreased renal cortical nitric oxide production and increasing renal injury, although the changes do not resemble those of diabetic nephropathy in man.Keywords Clearance · Creatinine · Endothelial nitric oxide synthase · Glomerulosclerosis · Hyperglycaemia · Kidney cortex · Neuronal nitric oxide synthase · Proteinuria · Type 2 diabetic nephropathy · Western blot Abbreviations: eNOS, endothelial nitric oxide synthase · ESRD, end-stage renal disease · nNOS, neuronal nitric oxide synthase · NOS, nitric oxide synthase · NO X , nitrite + nitrate ·

The Wistar-Furth (WF) rat is protected against chronic renal disease (CRD) following 5/6th ablation/infarction vs. the Sprague-Dawley (SD) rat, and protection was associated with preserved renal nitric oxide (NO) production. This study examined CRD induced with repeated administration of puromycin aminonucleoside (PAN). SD PAN developed nephrotic range proteinuria (>1 g/24 h), and at 15 wk severe renal injury developed and the glomerular filtration rate (GFR) was reduced to approximately 10% of sham. Total NO production, renal NO synthase (NOS) activity, and renal neuronal (n) and medullary endothelial (e)NOS abundance were reduced in the SD PAN. WF PAN exhibited less severe initial proteinuria (>400 mg/24 h), which abated within weeks, whereas GFR was normal and injury was minimal at 15 wk. Total NO production and renal NOS activity and abundance were significantly elevated compared with SD PAN. NOS mRNA (nNOS, eNOS, and inducible NOS) was not altered in WF, whereas SD showed significant increases in NOS gene expression with PAN. In conclusion, WF showed resistance to a second model of CRD with maintained renal NOS activity compared with SD.

Chronic nitric oxide synthase inhibition (NOSI) causes chronic kidney disease (CKD) in the Sprague Dawley (SD) rat. We previously showed that the Wistar-Furth (WF) rats are resistant to several models of CKD and maintain renal nitric oxide (NO) production compared with SD rats, whereas low-dose NOSI caused progression of CKD in WF rats. Here, we evaluate the impact of high-dose chronic NOSI in WF and SD rats, as well as intrarenal responses to an acute pressor dose of NOSI in the normal WF. Rats were given N(G)-nitro-l-arginine methyl ester (l-NAME) (150 and 300 mg/l for 6-10 wk) in the drinking water after an initial bolus tail vein injection. Both strains showed significant reductions in total NO production with chronic l-NAME. SD given 150 mg/l l-NAME for 6 wk developed proteinuria and renal injury, whereas WF rats receiving 150 mg/l l-NAME for 6-10 wk or 300 mg/l for 6 wk developed no proteinuria and minimal renal injury. Blood pressure was significantly elevated with chronic NOSI in both strains but was higher in the SD rat. There was little impact on renal nitric oxide synthase expression with l-NAME, except that cortical endothelial nitric oxide synthase abundance increased in WF after 6 wk (150 mg/l). Micropuncture experiments with acute pressor NOSI resulted in similar increases in systemic blood pressure in SD and WF rats, whereas WF rats showed a much smaller increment in glomerular blood pressure compared with SD rats. In conclusion, WF rats do not develop renal injury after chronic NOSI at, or above, a dose that causes significant injury in the SD rat. This protection may be associated with protection from glomerular hypertension.

ies show nitric oxide (NO) deficiency is both a cause and consequence of chronic kidney disease (CKD). Reduced renal neuronal NO synthase (nNOS) abundance and activity parallel development of CKD with different models in the Sprague-Dawley (SD) rats, whereas Wistar Furth (WF) rats are protected against CKD and show preserved renal NO production. In this study, we compared renal NO in response to DOCA/salt-induced injury between the WF and SD. Studies were conducted on sham WF (n ϭ 6) and SD (n ϭ 6) and uninephrectized (UNX)ϩ75 mg DOCAϩ1% NaCl (WF n ϭ 9; SD n ϭ 10) rats followed for 5 wk. Kidneys were harvested for Western blot, NOS activity, and histology. Other measurements included creatinine clearance and 24-h total NO production and urinary protein excretion. Absolute values of kidney weight were lower in WF than SD rats that showed similar percent increases with UNXϩDOCA/NaCl. Proteinuria and decreased creatinine clearance were present in the SD but not the WF rats following UNXϩDOCA/NaCl. Glomerular injury was mild in the WF compared with SD rats that showed many globally damaged glomeruli. Although renal nNOS abundance was decreased in both strains (higher baseline in WF), soluble NOS activity was maintained in the WF but significantly reduced in the SD rats. Renal endothelial NOS abundance and membrane NOS activity were unaffected by treatment. In summary, WF rats showed resistance to UNXϩDOCA/NaCl-induced CKD with maintained renal NO production despite mild reduction in nNOS abundance. Further studies are needed to evaluate how WF rats maintain renal NO production despite similar changes in abundance as the vulnerable SD strain. neuronal nitric oxide synthase; endothelial nitric oxide synthase; nitric oxide synthase activity; glomerulosclerosis; creatinine clearance RECENT EVIDENCE SUGGESTS that nitric oxide (NO) deficiency is both a cause and consequence of chronic kidney disease (CKD) (5). Clinical data show decreased total NO production in patients with CKD and end-stage renal disease (ESRD) (7,(21)(22)(23)31). In animal models of CKD, renal NO deficiency is evident irrespective of the initial insult (1, 9 -11, 20, 29, 30) and enhanced progression is seen with superimposed NO synthase (NOS) inhibition and protection with L-arginine supplementation (14, 16). Furthermore, chronic NOS inhibition alone leads to hypertension, proteinuria, and renal injury (33). Within the kidney, loss of the neuronal isoform of NOS always associates with injury in multiple models of CKD (9 -12, 20, 30) and correlates with level of damage and declining renal function (26).The Wistar Furth (WF) rat is remarkably resistant to CKD induced by chronic NOS inhibition, puromycin-aminonucleoside (PAN)-induced CKD, and 5/6th renal ablation/infarction (A/I) (8, 10, 12, 13) and total NO production and renal nNOS abundance were preserved compared with the progressing Sprague-Dawley (SD) rat (10, 12). Previous work showed that WF rats are resistant to mineralocorticoid-induced CKD (27). In this study, we used the uninephrectomy (UN...

Background: Plasma ADMA and oxidative stress are elevated in chronic kidney disease (CKD). We previously reported that 5/6 NX rats developed significant glomerulosclerosis which was attenuated by the antioxidant vitamin E (Vit E). The 5/6 NX group increased renal cortex NADPH‐dependent superoxide production vs. sham which was completely prevented by Vit E. Here we investigate the impact of 5/6 NX (by surgical excision of kidney tissue) on plasma ADMA and renal expression of PRMT1 and DDAH, as well as DDAH activity. We also investigated the impact of Vit E on ADMA and DDAH in 5/6 NX. Materials and Methods: We studied 3 groups of male SD rats: sham (n=6), 5/6 NX (n=6), and 5/6 NX treated with Vit E (2000 U/kg food) (n=5). Rats were sacrificed 15 wk after surgery. Results: Both 5/6 NX groups showed similar increases in plasma ADMA levels vs. sham. Renal cortex DDAH1 and DDAH2 protein expression was not different between 3 groups. However, the in vitro renal DDAH activity was decreased in 5/6 NX rats and this was not prevented by Vit E. Renal cortex PRMT1 expression was increased in 5/6 NX rats. Conclusions: ADMA levels rise in 5/6NX in association with increased PRMT protein and decreased DDAH activity in kidney. This reinforces the notion that kidney plays a significant role in regulation of ADMA levels. Unexpectedly, the antioxidant action of Vit E did not prevent the increase in ADMA and decrease in DDAH activity.