Renin Excretion Research Articles

Urinary excretion of renin and angiotensinogen in hypertensive children and adolescents.

IntroductionIn recent years hypertension has become an emerging condition in the young population. It has been proposed that the renin–angiotensin system plays an important role in regulation of blood pressure. We assessed whether activation of the intrarenal renin-angiotensin system occurs in hypertensive children and adolescents and what better reflects its activity: urine angiotensinogen (AGT) or urine renin (REN).Material and methodsThe study was conducted on a sample of 58 subjects with primary hypertension (HT) and 29 normotensive children and adolescents. We measured urine REN and AGT excretion and assessed the values in relation to blood pressure (BP) and other clinical parameters. Both REN and AGT values were calculated by urine creatinine: REN/cr. and AGT/cr., respectively.ResultsWe observed higher urine REN/cr. values in hypertensive subjects in comparison to the reference group (6.99 vs. 2.93, p = 0.003). Hypertensive participants showed positive correlations between urine REN/cr. and diastolic 24-hour BP (r = 0.42, p = 0.002) as well as between urine REN/cr. and urine AGT/cr. (r = 0.266, p = 0.044, respectively).ConclusionsIncreased urine REN/cr. in hypertensive children and adolescents and its positive correlation with BP may indicate its important role in the pathogenesis of HT. Perhaps urine REN/cr. could be a marker of intrarenal renin-angiotensin system activity. Nevertheless, further research should be undertaken to confirm this observation.

In PICU acute kidney injury stage 3 or mortality is associated with early excretion of urinary renin.

Urinary renin is proposed to be a novel prognostic biomarker of acute kidney injury (AKI) in adults. The intention of our study was to evaluate the early predictive value of urinary renin for AKI and pediatric intensive care unit (PICU) mortality in critically ill children. The first available urine sample during the first 24 h after admission was collected upon PICU admission for the measurement of renin using ELISA. Urinary renin concentrations were corrected for urinary creatinine (urinary renin-to-creatinine ratio, uRenCR). AKI was defined based on KDIGO criteria. Of the 207 children, 22 developed AKI, including 6 with stage 1, 6 with stage 2, and 10 with stage 3, and 14 died during PICU stay. There was a significant difference in uRenCR between non-AKI children and those with AKI stage 3 (P = 0.001), but not with AKI stage 1 or 2. The uRenCR remained associated with AKI stage 3 and PICU mortality after adjustment for potential confounders. The area under the receiver operating characteristic curve of uRenCR for discrimination of AKI stage 3 was 0.805, and PICU mortality was 0.801. Urinary renin was associated with the increased risk for AKI stage 3 and PICU mortality in critically ill children. Urinary renin is proposed to be a novel prognostic biomarker of AKI in adult patients. There are some differences between children and adults in physiological and pathophysiological characteristics. This study demonstrated that urinary renin was associated with the increased risk for AKI stage 3 and PICU mortality in critically ill children. Accurate identification of patients with severe renal injury or at high risk for mortality early in the disease course could augment the efficacy of available interventions and improve patient outcomes.

The evolving complexity of the collecting duct renin-angiotensin system in hypertension.

The intrarenal renin-angiotensin system is critical for the regulation of tubule sodium reabsorption, renal haemodynamics and blood pressure. The excretion of renin in urine can result from its increased filtration, the inhibition of renin reabsorption by megalin in the proximal tubule, or its secretion by the principal cells of the collecting duct. Modest increases in circulating or intrarenal angiotensin II (ANGII) stimulate the synthesis and secretion of angiotensinogen in the proximal tubule, which provides sufficient substrate for collecting duct-derived renin to form angiotensin I (ANGI). In models of ANGII-dependent hypertension, ANGII suppresses plasma renin, suggesting that urinary renin is not likely to be the result of increased filtered load. In the collecting duct, ANGII stimulates the synthesis and secretion of prorenin and renin through the activation of ANGII type 1 receptor (AT1R) expressed primarily by principal cells. The stimulation of collecting duct-derived renin is enhanced by paracrine factors including vasopressin, prostaglandin E2 and bradykinin. Furthermore, binding of prorenin and renin to the prorenin receptor in the collecting duct evokes a number of responses, including the non-proteolytic enzymatic activation of prorenin to produce ANGI from proximal tubule-derived angiotensinogen, which is then converted into ANGII by luminal angiotensin-converting enzyme; stimulation of the epithelial sodium channel (ENaC) in principal cells; and activation of intracellular pathways linked to the upregulation of cyclooxygenase 2 and profibrotic genes. These findings suggest that dysregulation of the renin-angiotensin system in the collecting duct contributes to the development of hypertension by enhancing sodium reabsorption and the progression of kidney injury.

Effect of sodium bicarbonate supplementation on the renin-angiotensin system in patients with chronic kidney disease and acidosis: a randomized clinical trial

BackgroundAcidosis-induced kidney injury is mediated by the intrarenal renin-angiotensin system, for which urinary renin is a potential marker. Therefore, we hypothesized that sodium bicarbonate supplementation reduces urinary renin excretion in patients with chronic kidney disease (CKD) and metabolic acidosis.MethodsPatients with CKD stage G4 and plasma bicarbonate 15–24 mmol/l were randomized to receive sodium bicarbonate (3 × 1000 mg/day, ~ 0.5 mEq/kg), sodium chloride (2 × 1,00 mg/day), or no treatment for 4 weeks (n = 15/arm). The effects on urinary renin excretion (primary outcome), other plasma and urine parameters of the renin-angiotensin system, endothelin-1, and proteinuria were analyzed.ResultsForty-five patients were included (62 ± 15 years, eGFR 21 ± 5 ml/min/1.73m2, plasma bicarbonate 21.7 ± 3.3 mmol/l). Sodium bicarbonate supplementation increased plasma bicarbonate (20.8 to 23.8 mmol/l) and reduced urinary ammonium excretion (15 to 8 mmol/day, both P < 0.05). Furthermore, a trend towards lower plasma aldosterone (291 to 204 ng/L, P = 0.07) and potassium (5.1 to 4.8 mmol/l, P = 0.06) was observed in patients receiving sodium bicarbonate. Sodium bicarbonate did not significantly change the urinary excretion of renin, angiotensinogen, aldosterone, endothelin-1, albumin, or α1-microglobulin. Sodium chloride supplementation reduced plasma renin (166 to 122 ng/L), and increased the urinary excretions of angiotensinogen, albumin, and α1-microglobulin (all P < 0.05).ConclusionsDespite correction of acidosis and reduction in urinary ammonium excretion, sodium bicarbonate supplementation did not improve urinary markers of the renin-angiotensin system, endothelin-1, or proteinuria. Possible explanations include bicarbonate dose, short treatment time, or the inability of urinary renin to reflect intrarenal renin-angiotensin system activity.Graphic abstract

Chios mastic gum decreases renin levels and ameliorates vascular remodeling in renovascular hypertensive rats.

Chios mastic gum (CMG) exerts robust anti-inflammatory and antioxidant properties and it affects pathways that are implicated in the pathophysiology of endothelial and vascular inflammation. Aim of this study was to test the hypothesis that CMG administration lowers blood pressure (BP) and improves hypertension-induced target organ damage. 2-kidney, 1-clip (2K1C) hypertensive rats were treated with CMG (40 mg/kg body weight/day) for 2-weeks after the establishment of hypertension. Acute CMG administration lowered systolic, diastolic and mean arterial BP, while these hemodynamic effects were sustained throughout the 2-week administration period. CMG group also exhibited alleviated target organ damage as proposed by amelioration of biomechanical properties of the aorta -including cross-sectional area (CSA), aortic wall stiffness and thickness-, reversal of myocardial small vessel hypertrophy and maintenance of serum albumin levels. The anti-hypertensive effects of CMG are likely to be mediated by the decrease in renin serum levels. Regression analysis indicated that the effect of CMG on organ damage was BP-lowering dependent and was not associated with direct effects of renin or with its anti-inflammatory properties. We suggest a BP lowering effect of CMG via down-regulation of renin excretion associated with attenuation of target organ damage and inflammatory status. These observations provide profound evidence for the beneficial role of CMG in hypertension, which could possibly translate to further clinical research.

ACUTE AND CHRONIC EFFECTS OF CHIOS MASTIC GUM ON BLOOD PRESSURE IN A RODENT MODEL OF 2-KIDNEY 1-CLIP HYPERTENSION

Objective: Chios mastic gum (CMG), a Greek plant resin, has been studied for its protective role against endothelial and vascular inflammation by suppressing oxidative stress and downregulating the expression of pro-inflammatory mediators. Objective: We sought to investigate the effect of CMG administration on blood pressure (BP) and hypertension-induced target organ damage. Design and method: 16-week-old male Wistar rats were allocated into 3 groups: Control group; 2-kidney, 1-clip (2K1C) group; CMG group which was treated with CMG (40 mg/kg body weight /day) for 2-weeks after the establishment of hypertension. Results: Acute CMG administration led to a decrease in systolic, diastolic and mean arterial BP (153 vs 188 mmHg, 108 vs 141 mmHg and 122 vs 156 mmHg for CMG and 2K1C groups, respectively), while these hemodynamic effects were sustained throughout the 2-week administration period (136 vs 194 mmHg, 100 vs 141 mmHg and 112 vs 159 mmHg for CMG and 2K1C groups, respectively. CMG also attenuated target organ damage as proposed by amelioration of biomechanical properties of the aorta –including cross-sectional area (CSA), aortic wall stiffness and thickness-, reversal of myocardial small vessel hypertrophy and maintenance of serum albumin levels. Furthermore, CMG administration was associated with decreased interleukin-6 (IL-6) and C-reactive protein (CRP) levels. The BP lowering effects of CMG are likely to be mediated by the decrease in renin serum levels. Regression analysis showed that the amelioration of organ damage was BP-lowering dependent and was not correlated with direct effects of renin or with its anti-inflammatory properties. Conclusions: Our results indicate an anti-hypertensive effect of CMG via down-regulation of renin excretion and suppression of the inflammatory cascade associated with alleviation of target organ damage. These observations provide substantial evidence for the potentially beneficial role of CMG in treating hypertension, yet clinical translation studies are further required.

Abstract P252: The Effects of Sodium Reduction on Metabolism and Thirst

Background: Recent studies challenge the traditional understanding of sodium physiology. Some animal studies suggest that high sodium intake may induce catabolism, leading to weight loss. Other studies suggest that high sodium intake might reduce, rather than increase, thirst. Hypothesis: Higher sodium intake increases thirst, fluid intake, and sodium excretion without altering energy intake or lowering weight. Methods: In the DASH-Sodium feeding study, adults with pre- or stage 1 hypertension without antihypertensive medications, were randomly assigned to the DASH diet or a control diet. On their assigned diet, participants consumed each of three sodium levels for 4 weeks (randomized crossover design). Participants were provided all meals, but could drink water freely. Throughout the trial, calorie intake was adjusted to keep weight constant. The three sodium levels (at 2100 kcal/d) were: low (1150 mg), medium (2300 mg), and high (3450 mg). Weight, energy intake, self-reported thirst, urine volume, plasma renin, serum aldosterone, urine osmolality, and urine sodium excretion were assessed at the end of each period. Results: Among 412 participants (57% women, 57% black, mean age 48 yrs), weight increased slightly with higher sodium on the control diet, but not the DASH diet; energy intake did not vary across sodium levels in both diets ( P -trends &gt; 0.34) ( Table ). In contrast, participants reported more thirst with high vs low sodium (both diets P -trends &lt; 0.001) and potentially higher fluid intake (urine volume) during the control diet (1,566 vs 1,491 ml on high vs low sodium; P -trend = 0.07). On both diets, plasma renin and serum aldosterone were lower with higher sodium (each P -trend &lt; 0.001). Likewise, both urine osmolality and sodium excretion were higher with higher sodium intake (all P -trends &lt; 0.001). Conclusions: Higher sodium intake did not alter energy intake, but did increase thirst and sodium excretion. These findings are consistent with the traditional understanding of the physiology of excess dietary sodium intake.

Urinary renin-angiotensin markers in polycystic kidney disease.

In autosomal dominant polycystic kidney disease (ADPKD), activation of the renin-angiotensin aldosterone system (RAAS) may contribute to hypertension and disease progression. Although previous studies have focused on circulating RAAS components, preliminary evidence suggests that APDKD may increase urinary RAAS components. Therefore, our aim was to analyze circulating and urinary RAAS components in ADPKD. We cross-sectionally compared 60 patients with ADPKD with 57 patients with non-ADPKD chronic kidney disease (CKD). The two groups were matched by sex, estimated glomerular filtration rate (eGFR), blood pressure, and RAAS inhibitor use. Despite similar plasma levels of angiotensinogen and renin, urinary angiotensinogen and renin excretion were five- to sixfold higher in ADPKD (P < 0.001). These differences persisted when adjusting for group differences and were present regardless of RAAS inhibitor use. In multivariable analyses, ADPKD, albuminuria, and the respective plasma concentrations were independent predictors for urinary angiotensinogen and renin excretion. In ADPKD, both plasma and urinary renin correlated negatively with eGFR. Total kidney volume correlated with plasma renin and albuminuria but not with urinary renin or angiotensinogen excretions. Albuminuria correlated positively with urinary angiotensinogen and renin excretions in ADPKD and CKD. In three ADPKD patients who underwent nephrectomy, the concentrations of albumin and angiotensinogen were highest in plasma, followed by cyst fluid and urine; urinary renin concentrations were higher than cyst fluid. In conclusion, this study shows that, despite similar circulating RAAS component levels, higher urinary excretions of angiotensinogen and renin are a unique feature of ADPKD. Future studies should address the underlying mechanism and whether this may contribute to hypertension or disease progression in ADPKD.

Sodium butyrate suppresses angiotensin II-induced hypertension by inhibition of renal (pro)renin receptor and intrarenal renin-angiotensin system.

Butyrate, a short-chain fatty acid, is the end product of the fermentation of complex carbohydrates by the gut microbiota. Recently, sodium butyrate (NaBu) has been found to play a protective role in a number of chronic diseases. However, it is still unclear whether NaBu has a therapeutic potential in hypertension. The present study was aimed to investigate the role of NaBu in angiotensin II (Ang II)-induced hypertension and to further explore the underlying mechanism. Ang II was infused into uninephrectomized Sprague-Dawley rats with or without intramedullary infusion of NaBu for 14 days. Mean arterial blood pressure was recorded by the telemetry system. Renal tissues, serum samples, and 24-h urine samples were collected to examine renal injury and the regulation of the (pro)renin receptor (PRR) and renin. Intramedullary infusion of NaBu in Sprague-Dawley rats lowered the Ang II-induced mean arterial pressure from 129 ± 6 mmHg to 108 ± 4 mmHg (P < 0.01). This corresponded with an improvement in Ang II-induced renal injury, including urinary albumin, glomerulosclerosis, and renal fibrosis, as well as the expression of inflammatory mediators tumor necrosis factor α, interleukin 6. The renal expression of PRR, angiotensinogen, angiotensin I-converting enzyme and the urinary excretion of soluble PRR, renin, and angiotensinogen were all increased by Ang II infusion but decreased by NaBu treatment. In cultured innermedullary collecting duct cells, NaBu treatment attenuated Ang II-induced expression of PRR and renin. These results demonstrate that NaBu exerts an antihypertensive action, likely by suppressing the PRR-mediated intrarenal renin-angiotensin system.

Urinary Angiotensinogen and Renin Excretion are Associated with Chronic Kidney Disease

Background/Aims: Several studies sought to identify new biomarkers for chronic kidney disease (CKD). As the renal renin-angiotensin system is activated in CKD, urinary angiotensinogen or renin excretion may be suitable candidates. We tested whether urinary angiotensinogen or renin excretion is elevated in CKD and whether these parameters are associated with estimated glomerular filtration rate (eGFR). We further tested whether urinary angiotensinogen or renin excretion may convey additional information beyond that provided by albuminuria. Methods: We measured urinary and plasma angiotensinogen, renin, albumin and creatinine in 177 CKD patients from the Greifswald Approach to Individualized Medicine project and in 283 healthy controls from the Study of Health in Pomerania. The urinary excretion of specific proteins is given as protein-to-creatinine ratio. Receiver operating characteristic (ROC) curves, spearman correlation coefficients and linear regression models were calculated. Results: Urinary angiotensinogen [2,511 (196-31,909) vs. 18.6 (8.3-44.0) pmol/g, *P<0.01] and renin excretion [0.311 (0.135-1.155) vs. 0.069 (0.045-0.148) pmol/g, *P<0.01] were significantly higher in CKD patients than in healthy controls. The area under the ROC curve was significantly larger when urinary angiotensinogen, renin and albumin excretion were combined than with urinary albumin excretion alone. Urinary angiotensinogen (ß-coefficient -2.405, standard error 0.117, P<0.01) and renin excretion (ß-coefficient -0.793, standard error 0.061, P<0.01) were inversely associated with eGFR. Adjustment for albuminuria, age, sex, systolic blood pressure and body mass index did not significantly affect the results. Conclusion: Urinary angiotensinogen and renin excretion are elevated in CKD patients. Both parameters are negatively associated with eGFR and these associations are independent of urinary albumin excretion. In CKD patients urinary angiotensinogen and renin excretion may convey additional information beyond that provided by albuminuria.

Collecting Duct Renin Does Not Mediate DOCA-Salt Hypertension or Renal Injury

Collecting duct (CD)-derived renin is involved in the hypertensive response to chronic angiotensin-II (Ang-II) administration. However, whether CD renin is involved in Ang-II independent hypertension is currently unknown. To begin to examine this, 12 week old male and female CD-specific renin knock out (KO) mice and their littermate controls were subjected to uni-nephrectomy followed by 2 weeks of deoxycorticosterone acetate (DOCA) infusion combined with a high salt diet. Radiotelemetric blood pressure (BP) was similar between KO and control mice at baseline; BP increased in both groups to a similar degree throughout the 2 weeks of DOCA-salt treatment. Urinary albumin excretion and plasma blood urea nitrogen were comparable between the two groups after DOCA-salt treatment. Fibrosis as assessed by Masson’s Trichrome stain/Sirius Red stain and collagen-1 mRNA expression was similar between control and KO mice. Compared to baseline, DOCA-salt treatment decreased plasma renin concentration (PRC), urinary renin excretion and medullary renin mRNA expression in both floxed and CD renin KO mice with no detectable differences between the two groups. Further, in primary culture of rat inner medullary CD, aldosterone treatment did not change renin activity or total renin content. Taken together, these data suggest that CD derived renin does not play a role in DOCA-salt hypertension.

Circulating and intrarenal renin-angiotensin systems in healthy men and nonpregnant women.

The urinary excretion of renin–angiotensin system (RAS) proteins could reflect the activity of the intrarenal RAS. We hypothesized that the rates of excretion of RAS components into human urine are independent of circulating levels of these proteins and reflect the intrarenal RAS. There are no reports of the simultaneous measurement of prorenin, active renin, angiotensinogen (AGT), and angiotensin-converting enzyme (ACE) excretion in healthy individuals. Therefore, we measured plasma prorenin, ACE, and AGT and urinary renin (uRenin), prorenin (uProrenin), ACE (uACE), and AGT (uAGT) in men and nonpregnant women. Plasma (p) AGT was higher in women then men. Women who were taking estrogen had significantly higher pAGT. In women, pProrenin was negatively correlated with pAGT. There were no correlations between pProrenin, pAGT, and pACE and their urinary counterparts in either men or women. In men, uProrenin/creatinine ratios were lower than in women. There was no effect of estrogen use on urinary excretion of pProrenin, renin, AGT, and ACE. In men, there were significant correlations between uACE/creat and uRen/creat and uAGT/creat; uProrenin/creat and plasma cystatin C levels; and uRenin/creat and uNa/K were also positively correlated. No associations were found in women. In conclusion, urinary excretion of prorenin is sexually dimorphic and is not affected by estrogen use in women. Our data also suggest that the relationship between renal handling of sodium and urinary renin is sexually dimorphic. Since we found no associations between plasma RAS proteins and their urinary counterparts, and the ratio of uProrenin:pProrenin was strikingly different between men and women, levels of urinary RAS proteins in individuals with normal kidney function are most likely the result of tubular secretion, rather than ultrafiltration.

Collecting duct-specific knockout of renin attenuates angiotensin II-induced hypertension.

The physiological and pathophysiological significance of collecting duct (CD)-derived renin, particularly as it relates to blood pressure (BP) regulation, is unknown. To address this question, we generated CD-specific renin knockout (KO) mice and examined BP and renal salt and water excretion. Mice containing loxP-flanked exon 1 of the renin gene were crossed with mice transgenic for aquaporin-2-Cre recombinase to achieve CD-specific renin KO. Compared with controls, CD renin KO mice had 70% lower medullary renin mRNA and 90% lower renin mRNA in microdissected cortical CD. Urinary renin levels were significantly lower in KO mice (45% of control levels) while plasma renin concentration was significantly higher in KO mice (63% higher than controls) during normal-Na intake. While no observable differences were noted in BP between the two groups with varying Na intake, infusion of angiotensin II at 400 ng·kg(-1)·min(-1) resulted in an attenuated hypertensive response in the KO mice (mean arterial pressure 111 ± 4 mmHg in KO vs. 128 ± 3 mmHg in controls). Urinary renin excretion and epithelial Na(+) channel (ENaC) remained significantly lower in the KO mice following ANG II infusion compared with controls. Furthermore, membrane-associated ENaC protein levels were significantly lower in KO mice following ANG II infusion. These findings suggest that CD renin modulates BP in ANG II-infused hypertension and these effects are associated with changes in ENaC expression.

Overexpression of Renin in the Collecting Duct Causes Elevated Blood Pressure

Renin is synthesized in the collecting duct and is regulated differently than renin in the juxtaglomerular apparatus. However, the physiological relevance of collecting duct renin remains unknown, particularly with regard to its ability to regulate blood pressure. We used gene targeting to generate mice with overexpression of renin in the collecting duct. A conditional mutant mouse line was created with the mouse renin transcript distal to a "transcriptional stop sequence" such that gene expression only occurred when the stop sequence was excised. These mice were bred with mice transgenic for the aquaporin-2 promoter driving Cre recombinase in order to achieve collecting duct-specific overexpression of renin. RNA analysis confirmed kidney-specific recombination, and medullary renin mRNA levels were increased 5-fold in collecting duct renin mice. Blood pressure was recorded by telemetry and plasma and urine was collected in 24-hour metabolic cages on normal, high-, and low-Na+ diets. Although no significant differences in 24-hour urinary Na+ excretion between targeted and control mice were detected, renin overexpresser mice had elevated blood pressure compared with controls on a high-Na+ diet. Urinary renin excretion was 2-fold higher in targeted mice as compared with controls on normal and low-Na+ diets. Plasma renin concentration was significantly suppressed in targeted mice as compared with controls on normal and high-Na+ diets. Taken together, these results suggest that collecting duct-derived renin has the potential to modulate blood pressure independent of the systemic renin-angiotensin system.

Overexpression of renin in the collecting duct causes hypertension

The role of collecting duct (CD)‐derived renin in regulating arterial pressure (BP) is unknown. To examine proof of concept, gene targeting was used to obtain mice with renin overexpression in the CD. Mice were generated in which the sequence: loxP ‐ PGK promoter‐NeoR ‐ transcriptional stop sequence – loxP ‐ Ren1 cDNA – bovine growth hormone polyA was knocked in to the ROSA26 locus. These mice were crossed with mice transgenic for aquaporin 2 AQP2) promoter‐Cre recombinase, resulting in excision of the PGK‐Neo‐Stop and CD‐specific overexpression of renin (CD renin mice). Mice heterozygous for the knock‐in allele and AQP2‐Cre were studied. BP was recorded by telemetry and plasma and urine were collected in metabolic cages on normal, high and low Na diets. DNA recombination and PCR for ROSA‐renin mRNA confirmed kidney‐specific expression. Medullary renin mRNA was increased 5‐fold and urinary renin excretion was increased 2.5‐fold in CD renin mice. CD renin mice had salt‐sensitive hypertension that was most apparent on a high Na diet (mean BP 105 ± 2 mmHg vs. 95 ± 2 mmHg in wild type). Plasma renin concentration (PRC) was reduced by ~50% and ~80% in CD renin mice on a normal or high Na diet, respectively, as compared to WT animals. There were no differences in BP or PRC between CD renin and WT mice fed a low Na diet. These findings indicate that CD‐derived renin has the potential to modulate BP and that this effect is due to local, and not systemic, renin activity.

The effects of calcitriol on albuminuria in patients with type-2 diabetes mellitus.

The renin-angiotensin system has a major role in the development of diabetic nephropathy (DN). It is reported that vitamin D analogues are able to suppress renin excretion. Thus, this study was conducted to determine whether there is any correlation between albuminuria as a marker of DN with vitamin D levels in diabetic patients. Also, an assessment was made on the effects of vitamin D therapy on albuminuria in this group of patients. We conducted this cross-sectional study on 119 outpatients with type-2 diabetes. The serum levels of 25-hydroxy vitamin D [25 (OH) D] and the albumin to creatinine ratio were assessed in all the study patients. Patients with vitamin D deficiency/insufficiency received calcitriol therapy for eight weeks, following which the laboratory tests were repeated. The mean age of the study patients was 55.3 ± 11.2 years, 43 (36.13%) had vitamin D insufficiency [25 (OH) D <25 ng/mL] and 31 (26.1%) had vitamin D deficiency [25 (OH) D <15 ng/mL]. We found a significant correlation between 25 (OH) D levels and presence of microalbuminuria (P = 0.04) in patients with vitamin D deficiency. Therapy with calcitriol had a beneficial effect on the albumin excretion rate, although this change was not significant (P = 0.22). However, the effects of calcitriol on reduction of diastolic blood pressure (P = 0.004), glycosylated hemoglobin (P = 0.014) and levels of total cholesterol (P = 0.019), low-density lipoprotein (0.04) and high-density lipoprotein (P = 0.001) was significant. Our study suggests that vitamin D deficiency has a negative effect on albuminuria in diabetic patients, and its replacement may be associated with a beneficial effect on the risk factors of DN, such as hyperlipidemia and hypertension.

Abstract P264: Interaction of Collecting Duct-Derived Prorenin and Soluble Prorenin Receptor Increases Intraluminal Renin Activity and Augments Intratubular Angiotensin II Formation in Ang II-Dependent Hypertensive Rats

Upregulation of collecting duct (CD)-derived renin (CD renin) in angiotensin II (Ang II)-dependent hypertension may provide a pathway for intratubular Ang II formation by acting on angiotensinogen (AGT) delivered from proximal tubule segments. Recently, a prorenin/renin receptor (PRR) has been cloned and shown to enhance renin and prorenin activation. The soluble form of the PRR (sPRR) is augmented in the renal inner medulla of chronic Ang II-infused rats. The present study was performed to determine if renin is secreted into the lumen by the CD cells in chronic Ang II-infused rats and to establish the functional contribution of sPRR to the enhanced renin activity in distal nephron segments. Accordingly, urinary levels of renin ( uRen ) and Ang II ( uAngII ) were measured by RIA in chronic Ang II-infused male Sprague-Dawley rats [80 ng/min, SC minipumps for 14 d, n=10] and sham-operated rats [n=10]. Systolic blood pressure increased in the Ang II rats by Day 5 and continued to increase throughout the study (Day 13; Ang II: 175±10 vs. sham: 116±2 mmHg; p &lt;0.05). Although plasma renin activity (PRA) was suppressed in the Ang II-infused rats, renal medullary renin content was significantly augmented (12,605±1,343 vs. 7,956±765 ng Ang I/h/mg; p &lt;0.05). The excretion of uAngII was also increased (3,813±431 vs. 2,080±361 fmol/day; p &lt;0.05). In addition, renin and prorenin excretion rates increased progressively and were markedly augmented by Day 13 of Ang II infusion [renin (8.6±1.5 vs. 2.8±0.5x10 -6 Enzyme Units Excreted (EUE) /day; prorenin: 15.8 ± 2.8 vs. 2.6 ± 0.7x10 -3 EUE /day, p &lt;0.05). Renin and prorenin protein levels examined by Western Blot in the urine were similarly increased. Importantly, the incubation of urine samples of Ang II-infused rats with recombinant human prorenin showed increased Ang I formation compared to sham-operated rats. In conclusion, in chronic Ang II-infused rats, the presence of sPRR in the urine reflects augmented enzymatic activity of prorenin secreted by the principal cells of the CD, which increase intratubular Ang II de novo formation in the distal nephron segments thus contributing to enhanced sodium reabsorption during Ang II-dependent hypertension.

Urinary renin, but not angiotensinogen or aldosterone, reflects the renal renin–angiotensin–aldosterone system activity and the efficacy of renin–angiotensin–aldosterone system blockade in the kidney

To study which renin-angiotensin-aldosterone system (RAAS) component best reflects renal RAAS activity. We measured urinary and plasma renin, prorenin, angiotensinogen, aldosterone, albumin and creatinine in 101 diabetic and nondiabetic patients with or without hypertension. Plasma prorenin was elevated in diabetic patients. Urinary prorenin was undetectable. Urinary albumin and renin were higher in diabetic patients. Men had higher plasma renin/prorenin levels, and lower plasma angiotensinogen levels than women. Plasma creatinine and albumin were also higher in men. Urinary RAAS components showed no sexual dimorphism, whereas urinary creatinine and albumin were higher in men. Angiotensin-converting enzyme inhibitors and angiotensin II type 1 receptor blockers increased plasma renin and decreased plasma angiotensinogen, without altering plasma aldosterone. In contrast, in urine, these drugs decreased renin and aldosterone without affecting angiotensinogen. When analyzing all patients together, urinary angiotensinogen excretion closely mimicked that of albumin, whereas urinary angiotensinogen and albumin levels both were 0.05% or less of their concomitant plasma levels. This may reflect the identical glomerular filtration and tubular handling of both proteins, which have a comparable molecular weight. In contrast, urinary renin excretion did not correlate with urinary albumin excretion, and the urinary/plasma concentration ratio of renin was more than 200 times the ratio of albumin, despite its comparable molecular weight. Urinary aldosterone excretion closely followed urinary creatinine excretion. The increased urinary renin levels in diabetes and the decreased urinary renin levels following RAAS blockade, occurring independently of changes in plasma renin, reflect the activated renal RAAS in diabetes and the success of RAAS blockade in the kidney, respectively. Urinary renin, therefore, more closely reflects renal RAAS activity than urinary angiotensinogen or aldosterone.

Increased renin excretion is associated with augmented urinary angiotensin II levels in chronic angiotensin II-infused hypertensive rats

Renin expression in principal cells of collecting ducts (CD) is upregulated in angiotensin II (ANG II)-dependent hypertensive rats; however, it remains unclear whether increased CD-derived renin undergoes tubular secretion. Accordingly, urinary levels of renin (uRen), angiotensinogen (uAGT), and ANG II (uANG II) were measured in chronic ANG II-infused Sprague-Dawley rats (80 ng/min for 14 days, n = 10) and sham-operated rats (n = 10). Systolic blood pressure increased in the ANG II rats by day 5 and continued to increase throughout the study (day 13; ANG II: 175 ± 10 vs. sham: 116 ± 2 mmHg; P < 0.05). ANG II infusion increased renal cortical and medullary ANG II levels (cortical ANG II: 606 ± 72 vs. 247 ± 43 fmol/g; P < 0.05; medullary ANG II: 2,066 ± 116 vs. 646 ± 36 fmol/g; P < 0.05). Although plasma renin activity (PRA) was suppressed in the ANG II-infused rats (0.3 ± 0.2 vs. 5.5 ± 1.8 ng ANG I·ml(-1)·h(-1); P < 0.05), renin content in renal medulla was increased (12,605 ± 1,343 vs. 7,956 ± 765 ng ANG I·h(-1)·mg(-1); P < 0.05). Excretion of uAGT and uANG II increased in the ANG II rats [uAGT: 1,107 ± 106 vs. 60 ± 26 ng/day; P < 0.0001; uANG II: 3,813 ± 431 vs. 2,080 ± 361 fmol/day; P < 0.05]. By day 13, despite suppression of PRA, urinary prorenin content increased in ANG II rats [15.7 ± 3 vs. 2.6 ± 1 × 10(-3) enzyme units excreted (EUE)/day, P < 0.01] as was the excretion rate of renin (8.6 ± 2 × 10(-6) EUE/day) compared with sham (2.8 ± 1 × 10(-6) EUE/day; P < 0.05). Urinary renin and prorenin protein levels examined by Western blot were augmented ∼10-fold in the ANG II-infused rats. Concomitant AT(1) receptor blockade with candesartan prevented the increase. Thus, in ANG II-dependent hypertensive rats with marked PRA suppression, increased urinary levels of renin and prorenin reflect their augmented secretion by CD cells into the luminal fluid. The greater availability of renin and AGT in the urine reflects the capability for intratubular ANG II formation which stimulates sodium reabsorption in distal nephron segments.

Increased Urinary Renin Excretion Rate in Chronic Ang II‐infused Rats Fed a High Salt Diet leads to augmented urinary Ang II levels.

This study examined the urinary excretion of renin (uRen), angiotensinogen (uAGT), and angiotensin II (uAngII) in chronic AngII‐infused Sprague‐Dawley rats fed a high salt (HS) diet. Three groups were studied: 1) AngII‐infused [80 ng/min, 14 d, n=5]; 2) AngII‐infused plus HS [Ang II+HS; 8% NaCl; n=4]; 5) AngII‐infused, HS plus an AngII type 1 receptor blocker (ARB) [Ang II+HS+Candesartan, 25 mg/L, drinking water, 14 d, n=5]. Systolic blood pressures increased in AngII (175±10 mmHg) and were augmented by HS in AngII+HS rats (222±8 mmHg). By day 13, uRen, uAGT and uAngII excretion rates increased in AngII‐infused rats, and in AngII+HS rats: [uRen (AngII: 9±3 vs. AngII+HS: 10±4 ×10‐5 Enzyme Units/day); uAGT (AngII: 1107±106 vs. AngII+HS: 4363±1673 ng/day); uAngII: (HS:1,851±327; AngII: 3,813±431; AngII+HS: 8,570±889 vs. NS: 2080±361 fmol/day; respectively)]. ARB treatment prevented these increases (uRen: 5±3×10‐5 Enzyme Units/day; uAGT: 240±28 ng/day; uAngII: 3,796±1,059 fmol/day). Thus, HS exacerbates the increases in uRen, uAGT, and uAngII as well as the systolic blood pressure in AngII‐dependent hypertensive rats.NIH [IdeA Program (P20‐RR‐017659‐06) and the Tulane‐BIRCWH Program (K12HD043451)]; and the AHA (09PRE2209079 and 09BGIA2280440).