Posttranslational regulation of NO synthase activity in the renal medulla of diabetic rats (original) (raw)
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Diabetologia, 2000
Several investigations have shown that the renal medulla has a greater capacity to generate nitric oxide than the renal cortex. To further evaluate the changes of nitric oxide synthesis in the kidney, particularly in the outer medulla, in disorders involving fluid and electrolyte imbalances, we sought to determine renal nitric oxide synthase expression in the diabetic rats. We determined renal nitric oxide synthase mRNA and urinary nitrite/nitrate excretion in 12 normal and 12 streptozotocin-induced diabetic rats by reverse transcription-polymerase chain reaction with Southern blot hybridization and with Griess reaction, respectively. Nitric oxide synthase immunoreactivity was detected by immunohistochemistry in four normal and four diabetic rats. Neuronal and endothelial nitric oxide synthase mRNA were 3.5-fold and 1.8-fold increased in the outer medulla of 12 diabetic rats with no difference found in the cortex and inner medulla when compared with 12 normal rats. Urinary nitrite/nitrate excretion was significantly increased from the first week after diabetic induction. In normal rats, immunohistochemical studies showed positive neuronal and endothelial nitric oxide synthase immunostaining in almost all segments of renal tubules. Diabetic rats had the greatest enhancement of immunostaining for neuronal and endothelial nitric oxide synthase in the proximal straight tubule and medullary thick ascending limb. Our results indicate that increases in neuronal and endothelial nitric oxide synthase synthesis in the kidney, particularly in the outer medulla, possibly play an important part in the adaptation of renal function to hyperglycaemia and hyperosmolality in diabetes.
Chronic diabetic nephropathy: role of inducible nitric oxide synthase
Pediatric Nephrology, 2002
Nitric oxide (NO) is a multifunctional mediator that has been implicated in the short-term hemodynamic alterations that occur in acute streptozocin (STZ)-induced diabetes. We investigated the role of NO produced by inducible nitric oxide synthase (iNOS) in chronic STZ diabetic nephropathy. Diabetes was induced in C57BL/6 and iNOS knockout (KO) mice with two intraperitoneal injections of STZ, 100 mg/kg. Animals were maintained without insulin treatment for 40 weeks. There were no significant differences between the strains in blood urea nitrogen (BUN), serum creatinine or glucose concentration, or urinary protein excretion during the entire observation period. Urinary nitrite + nitrate excretion was significantly lower in iNOS KO mice compared to control animals at all time points; in C57 mice, urinary nitrite declined progressively with more prolonged duration of diabetes. Renal hypertrophy (kidney weight/body weight) was noted in both strains of mice. However, histopathological assessment of renal tissue specimens at 16 and 40 weeks demonstrated increased mesangial hypercellularity and expansion as well as more prominent tubulointerstitial fibrosis in iNOS KO versus C57 mice. These changes were accompanied by increased interstitial deposition of type I collagen at 16 and 40 weeks in iNOS KO mice. Glomerular basement membrane staining for type IV collagen was also increased at 40 weeks in diabetic iNOS KO mice. While iNOS protein was undetectable in any of the kidney specimens obtained from either strain, eNOS was present throughout the course of chronic STZ diabetes. Moreover, eNOS expression was significantly increased by approximately 40% at 16 and 40 weeks of observation in iNOS KO versus C57 mice. There was no difference in renal cortical malondialdehyde content between the strains early or late in the disease course. In time control animals, there was no evidence of renal histopathological damage in iNOS KO or C57 mice after 40 weeks. We conclude that iNOS-derived NO modulates glomerulosclerosis and tubulointerstitial fibrosis in chronic STZ nephropathy. This action is probably a result of the direct actions of NO on the synthesis and degradation of extracellular matrix proteins.
Diabetes, Obesity and Metabolism, 1999
Objective: Nitric oxide (NO) has been proposed to play a signi®cant role in renal function. In addition, NO production has been found to increase in diabetes mellitus. The present study aimed to clarify the mechanism responsible for NO action in renal function in rats with short (10 days) or prolonged periods (8 weeks) of diabetic induction. Methods: Male Wistar rats were induced to develop diabetes mellitus by intraperitoneal injection of streptozotocin (STZ) (65 mg/kg b.w.), whereas the age-matched control rats were given normal saline. After diabetic induction for 10 days or 8 weeks, the experiment was begun. Three consecutive periods of 30 min each, were designed consisting of one control period, the ®rst and the second period of L-arginine or L-NAME or insulin infusion. Mean arterial pressure (MAP) was determined every 15 min. Arterial blood and urine samples were collected to determine the plasma glucose level (PG), glomerular ®ltration rate (GFR), effective renal plasma¯ow (ERPF), urine¯ow rate (V), urinary protein excretion (Upro), fractional excretion of glucose (FEG) and fractional excretion of sodium (FENa) in each period. Results: No signi®cant differences of MAP were apparent between control rats and rats with diabetic induction. L-arginine infusion had no effect whereas L-NAME markedly increased MAP in normal rats and rats after the short period of diabetes induction. Pressor response to L-NAME in rats exposed to the prolonged period of diabetes induction was lower than that of age-matched control rats. During L-NAME infusion, the PG level signi®cantly declined from 394.9 6 13.1± 338.0 6 14.1 mg/dl and from 399.9 6 7.9±354.3 6 18.8 mg/dl in rats after short and prolonged periods of diabetic induction, respectively. GFR signi®cantly increased whereas ERPF slightly increased in diabetic rats. The elevation of GFR could be reversed by L-NAME or insulin infusion but it increased again after simultaneous infusion of insulin and glucose. Increases in V, the Upro and FEG without changes of FENa, were apparent in diabetic rats. Either L-arginine or L-NAME infusion could not reverse elevations of V, Upro and FEG. The rise of both V and Upro was reversed along with the attenuation of high FEG during insulin infusion, and it rose again close to the diabetic level during simultaneous infusion of insulin and glucose. Elevation of GFR, V and Upro appeared along with a rise of the PG level by » 300±350 mg/ dl in diabetic rats. Conclusions: Both NO and hyperglycaemia are involved in modulating renal hyper®ltration in diabetic rats. The elevations of urine¯ow rate and urinary excretion of both protein and glucose would be expected to represent the reduction of renal tubular reabsorption rather than renal hyper®ltration in diabetic rats. NO does not participate in the change of renal tubular function in diabetic rats. There was a parallel change of urine¯ow rate and urinary excretion of protein in diabetic rats. The rise of the PG level itself would account for the increases of GFR, V, Upro and FEG in diabetic rats. Glomerular hyper®ltration, diuresis and proteinuria in diabetic rats are not exhibited until the PG level rises to » 300±350 mg/ dl.
American Journal of Physiology-Renal Physiology, 2019
Na+-glucose cotransporter (SGLT)1 mediates glucose reabsorption in late proximal tubules. SGLT1 also mediates macula densa (MD) sensing of an increase in luminal glucose, which increases nitric oxide (NO) synthase 1 (MD-NOS1)-mediated NO formation and potentially glomerular filtratrion rate (GFR). Here, the contribution of SGLT1 was tested by gene knockout (−/−) in type 1 diabetic Akita mice. A low-glucose diet was used to prevent intestinal malabsorption in Sglt1−/−mice and minimize the contribution of intestinal SGLT1. Hyperglycemia was modestly reduced in Sglt1−/−versus littermate wild-type Akita mice (480 vs. 550 mg/dl), associated with reduced diabetes-induced increases in GFR, kidney weight, glomerular size, and albuminuria. Blunted hyperfiltration was confirmed in streptozotocin-induced diabetic Sglt1−/−mice, associated with similar hyperglycemia versus wild-type mice (350 vs. 385 mg/dl). Absence of SGLT1 attenuated upregulation of MD-NOS1 protein expression in diabetic Akita...
Role of Renal Nitric Oxide Synthase in Diabetic Kidney Disease during the Chronic Phase of Diabetes
Nephron Physiology, 2006
Background: Several studies have suggested that an early increase in renal nitric oxide (NO) production or activity mediates pathophysiologic and morphologic changes in diabetic nephropathy. To evaluate the role of NO in developing diabetic kidney disease, we studied the NO system in streptozotocin (STZ)-induced diabetic rats for a period of 8 weeks. Methods: Control rats, STZ-induced diabetic rats, and STZ-induced diabetic rats treated with insulin were monitored and sacrificed at 1, 2, and 8 weeks. Urinary cGMP was measured, and the levels and activity of NO synthase (NOS) isoforms in the kidney cortex were determined at specific times by immunoblotting and diaphorase staining. Results: Diabetic rats had increased kidney weight, urinary volume, glucose, sodium and potassium excretion, which was precluded by insulin treatment. Creatinine clearance was increased in the diabetic group and reversed by insulin treatment. Urinary cGMP decreased by 71, 93, and 92% at 1, 2, and 8 weeks of...
Nitric oxide system and diabetic nephropathy
Diabetology & Metabolic Syndrome, 2014
About 30% of patients with type 2 diabetes mellitus develop clinically overt nephropathy. Hyperglycemia is necessary, but not sufficient, to cause the renal damage that leads to kidney failure. Diabetic nephropathy (DN) is a multifactorial disorder that results from interaction between environmental and genetic factors. In the present article we will review the role of the nitric oxide synthase (NOS) in the pathogenesis of DN. Nitric oxide (NO) is a short-lived gaseous lipophilic molecule produced in almost all tissues, and it has three distinct genes that encode three NOS isoforms: neuronal (nNOS), inducible (iNOS) and endothelial (eNOS). The correct function of the endothelium depends on NO, participating in hemostasis control, vascular tone regulation, proliferation of vascular smooth muscle cells and blood pressure homeostasis, among other features. In the kidney, NO plays many different roles, including control of renal and glomerular hemodynamics. The net effect of NO in the kidney is to promote natriuresis and diuresis, along with renal adaptation to dietary salt intake. The eNOS gene has been considered a potential candidate gene for DN susceptibility. Three polymorphisms have been extensively researched: G894T missense mutation (rs1799983), a 27-bp repeat in intron 4, and the T786C single nucleotide polymorphism (SNP) in the promoter (rs2070744). However, the potential link between eNOS gene variants and the induction and progression of DN yielded contradictory results in the literature. In conclusion, NOS seems to be involve in the development and progression of DN. Despite the discrepant results of many studies, the eNOS gene is also a good candidate gene for DN.
Glucose stimulates O2 consumption, NOS, and Na/H exchange in diabetic rat proximal tubules
American Journal of Physiology-Renal …, 2002
Endothelial nitric oxide synthase (NOS) and neuronal NOS protein increased in proximal tubules of acidotic diabetic rats 3-5 wk after streptozotocin injection. NOS activity (citrulline production) was similar in nondiabetic and diabetic tubules incubated with low glucose (5 mM glucose ϩ 20 mM mannitol); but after 30 min with high glucose (25 mM), Ca-sensitive citrulline production had increased 23% in diabetic tubules. Glucose concentration did not influence citrulline production in nondiabetic tubules. High glucose increased carboxy-2-phenyl-4,4,5,5,-tetramethylimidazoline 1-oxyl-3-oxide (cpt10)-scavenged NO sevenfold in a suspension of diabetic tubules but did not alter NO in nondiabetic tubules. Diabetes increased ouabain-sensitive 86 Rb uptake (141 Ϯ 9 vs. 122 Ϯ 6 nmol ⅐ min Ϫ1 ⅐ mg Ϫ1) and oligomycin-sensitive O 2 consumption (Q O2; 16.0 Ϯ 1.7 vs. 11.3 Ϯ 0.7 nmol ⅐ min Ϫ1 ⅐ mg Ϫ1). Ethylisopropyl amiloride-inhibitable Q O2 (6.5 Ϯ 0.6 vs. 2.4 Ϯ 0.3 nmol ⅐ min Ϫ1 ⅐ mg Ϫ1) accounted for increased oligomycin-sensitive Q O2 in diabetic tubules. N Gmonomethyl-L-arginine methyl ester (L-NAME) inhibited most of the increase in 86 Rb uptake and Q O2 in diabetic tubules. L-NAME had little effect on nondiabetic tubules. Inhibition of Q O2 by ethylisopropyl amiloride and L-NAME was only 5-8% additive. Uncontrolled diabetes for 3-5 wk increases NOS protein in proximal tubules and makes NOS activity sensitive to glucose concentration. Under these conditions, NO stimulates Na-K-ATPase and Q O2 in proximal tubules.
CHARACTERIZATION AND LOCALIZATION OF NITRIC OXIDE SYNTHASE ISOFORMS IN THE BB/WOR DIABETIC RAT
The Journal of Urology, 2001
Purpose: Erectile dysfunction is a common pathological development in individuals with diabetes mellitus. Nitric oxide synthase (NOS) is essential for regulation of normal penile erection and NOS protein activity has been shown to be altered with diabetes. Several different isoforms and subtypes of NOS exist. However, little is known about how the distribution and abundance of these isoforms are altered with diabetes. We characterized the distribution and abundance of NOS isoforms and explored how they are altered with diabetes and result in erectile failure.
Role of nitric oxide and angiotensin II in diabetes mellitus-induced glomerular hyperfiltration
Journal of the American Society of Nephrology, 1996
The goal of this study was to determine what extent nitric oxide (NO) and/or angiotensin II (AngII) are involved in the hyperfiltration observed in rats with streptozotocin-induced diabetes mellitus. Studies were performed on anesthetized rats 7 to 10 days after the induction of diabetes. Nitro-L-arginine (LNA) was used to inhibit NO synthesis, and losartan was used to block AngII receptors. Three protocols were utilized: (i) control and diabetic rats treated with a constant infusion of LNA; (ii) control and diabetic rats treated first with a constant infusion of losartan and then LNA plus losartan; and (iii) nephrectomized control and diabetic rats treated with LNA (to evaluate the involvement of renal vasoactive factors other than AngII in the systemic response to LNA). Compared with controls, diabetics had a significantly elevated baseline GFR but the same mean arterial pressure (MAP). In Protocol i, LNA caused the same increase in MAP in both groups but only decreased the GFR in...