Daniel Casellas

Preglomerular Sudanophilia in L-NAME Hypertensive Rats: Involvement of Endothelin

To characterize alterations of renal vessels occurring during systemic hypertension elicited in rats by 5, 10, and 25 days of treatment by the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME)(20 mg/kg daily), preglomerular vasculatures, consisting of arcuate arteries and their branches, interlobular arteries, and afferent arterioles, were isolated by HCl maceration. Blockade of nitric oxide synthase significantly increased tail-cuff systolic blood pressure by 21 +/- 2% and 42 +/- 3% after 5 and 25 days, respectively. Medias of hypertensive arcuate arterial branches and interlobular arteries but not of afferent arterioles had focal deposits of Sudan black-positive lipid droplets. At 25 days, vessel wall thickness increased by 72 +/- 6% along the sudanophilic areas. Immunostaining of sudanophilic lesions with a panel of antibodies unveiled medial cell proliferation, macrophage invasion, immunoreactive vascular cell adhesion molecule-1, and low-density lipoprotein. The frequency of sudanophilic lesions increased with time to affect 26 +/- 2% and 36 +/- 3% of arcuate arterial branches and interlobular arteries, respectively, at 25 days. Hypertensive L-NAME-treated rats developed glomerular injury probed by albuminuria and glomerular immunostaining for alpha-smooth muscle actin. Administration of the nonselective endothelin antagonist bosentan (30 mg/kg daily) blunted the development of sudanophilic lesions during L-NAME treatment without affecting arterial hypertension or degree of glomerular injury. Therefore, L-NAME hypertension leads to rapid development of focal, inflammatory, proliferative, and sudanophilic lesions along preglomerular vessels, suggesting atherosclerosis-like processes. Furthermore, endothelin is a likely mediator in the development of these lesions.

Pioglitazone Improves Aortic Wall Elasticity in a Rat Model of Elastocalcinotic Arteriosclerosis

Specific treatment of age-related aortic wall arteriosclerosis and stiffening is lacking. Because ligands for peroxisome proliferator–activated receptor &ggr; have beneficial effects on the arterial wall in atherosclerosis, via an antiinflammatory mechanism, we investigated whether long-term pioglitazone (Pio) treatment protects against another form of vascular wall disease, arteriosclerosis. We evaluated, in a rat model of elastocalcinotic arteriosclerosis (hypervitaminosis D and nicotine [VDN]), whether Pio (3 mg · kg−1 per day for 1.5 month PO) attenuated arteriosclerosis and its consequences: aortic wall rigidity, increased aortic pulse pressure, and left ventricular hypertrophy. In VDN rats, medial calcification was associated with monocyte/macrophage infiltration and induction of tumor necrosis factor &agr; and interleukin 1&bgr;. Pio increased nuclear peroxisome proliferator–activated receptor &ggr; immunostaining in the aortic wall, decreased tumor necrosis factor &agr; (P<0.05 versus VDN Pio−), tended to decrease interleukin 1&bgr; mRNA expression (P=0.08 versus VDN Pio−), blunted aortic wall calcification (271±69, P<0.05 versus VDN Pio− 562±87 &mgr;mol · g−1 dry weight) and prevented fragmentation of elastic fibers (segments per 10 000 &mgr;m2: 8.4±0.3; P<0.05 versus VDN Pio− 10.5±0.6). Pio reduced aortic wall stiffness (elastic modulus/wall stress: 4.8±0.6; P<0.05 versus VDN Pio− 10.0±1.6), aortic pulse pressure (30±2 mm Hg; P<0.05 versus VDN Pio− 39±4) and left ventricular hypertrophy (1.58±0.05 g · kg−1; P<0.05 versus VDN Pio− 1.76±0.06). In conclusion, long-term Pio treatment attenuates aortic wall elastocalcinosis and, thus, lowers aortic wall stiffness, aortic pulse pressure, and left ventricular hypertrophy.

Bosentan Prevents Preglomerular Alterations During Angiotensin II Hypertension

The present study was performed to characterize structurofunctional alterations of preglomerular vessels during chronic angiotensin II (Ang II)-induced hypertension (Ang II group: 400 ng x kg[-1] x min[-1], 10 days) and to assess the role of endothelin-1 in rats receiving Ang II and the mixed receptor antagonist bosentan (Ang II+B group: 30 mg x kg[-1] x d[-1], 10 days). Systolic blood pressure rose by 56+/-3 and 54+/-6 mm Hg in Ang II and Ang II+B rats, respectively. Albuminuria increased similarly in both Ang II-treated groups, reflecting glomerular barrier dysfunction. Preglomerular vessels were isolated after HCI maceration and comprised arcuate arteries and their branches, interlobular arteries (ILA), and afferent arterioles (AA). In the Ang II group, focal vascular lesions affected 36+/-6%, 20+/-5%, and 4+/-1% of arcuate arterial branches, ILA, and AA, respectively. They were characterized by 74% increased media thickness and accumulation of Sudan black-positive (SB+) lipid droplets, and media cell proliferation was documented through immunohistochemistry. The occurrence of SB+ lesions was strikingly reduced with bosentan. Autoregulatory responses (AR) were assessed along ILA and AA with the use of blood-perfused juxtamedullary nephron preparations. AR were elicited by raising blood perfusion pressure from 60 to 160 mm Hg and quantified through videomicroscopy as pressure-induced constrictions. AR were inhibited in Ang II-treated rats along ILA and AA; Ang II-induced AR changes were prevented by bosentan. Maximal relaxation induced by Mn2+ revealed equal basal tone in Ang II-treated, Ang II+B-treated, and control vessels. Chronic Ang II-induced hypertension is therefore associated with the development of SB+ lesions and selective impairment of AR in juxtamedullary nephrons. Endothelin-1 likely mediates the structurofunctional alterations of preglomerular vasculature during Ang II hypertension.

Renal adaptation to sodium deprivation. Effect of captopril in the rat.

Dietary sodium restriction is associated with a rapid decrease in urinary sodium excretion and achievement of a new sodium balance within three to five days. In addition, renal vasoconstriction and progressive activation of intrarenal systems with vasoconstrictor (renin-angiotensin) or vasodilating (kallikrein-kinin and prostaglandins) properties are observed. The relationship between sodium homeostasis and the renin-angiotensin system was assessed through the use of captopril in the rat. Treatment with captopril, before and during a six-day period after suppression of dietary sodium, was associated with sodium wasting (urinary sodium always exceeded sodium intake during the observation period); in addition, the normal increase in urinary aldosterone was blunted by about 80 percent. When captopril treatment was given for six days to rats maintained on long-term sodium restriction (at least four weeks) urinary sodium increased, although transiently; at the end of the study, renal vasodilatation together with a redistribution of glomerular blood flow to nonsuperficial glomeruli was observed. These studies indicate that captopril administration markedly blunts the renal and systemic adaptations to a reduced sodium intake in the rat. They suggest that the renin-angiotensin system is probably indispensable in preventing sodium loss when dietary sodium is suppressed.

Topology of Schwann cells and sympathetic innervation along preglomerular vessels: a confocal microscopic study in protein S100B/EGFP transgenic mice.

Schwann cells (Sc), associated axons, and nearby vascular endothelium constitute a functional trilogy of major importance during the development and regrowth of peripheral vascular nerves. The goal of the present study is to provide a technique of triple fluorescence confocal imaging of these cell types along renal preglomerular vessels. We took advantage of a protein S100B/EGFP transgenic mouse to visualize Sc. The endothelium was labeled with an intravenous injection of fluorescently tagged lectin, and after tissue processing, adrenergic nerves were revealed with an antibody against the marker protein synaptophysin. As a validation step, we found that EGFP-positive perivascular cells with prominent cell bodies and extensive, multidirectional cell processes were protein S100B positive. They were identified as Sc and indirectly assumed to be unmyelinated Sc. By contrast, we found strong EGFP expression in proximal epithelial cells and in the epithelium lining thin limbs of Henle. This epithelial fluorescence was not associated with immunoreactive protein S100B and thus corresponded to ectopic EGFP expressions in this mouse strain. Sc were organized in bundles or as a meshwork surrounding the preglomerular vasculature from arcuate arteries to afferent arterioles. No Sc were detected in the medulla. Although most Sc were closely apposed to adrenergic varicosities, many varicosities were not associated with detectable Sc processes. The present technique, and the capacity of confocal microscopy to yield three-dimensional imaging, allow the study of the microtopology of Sc and related sympathetic axons in the renal perivascular interstitium.

Influence of irbesartan and enalapril on changes of renal function associated with the established phase of l-NAME hypertension.

Objective Reversibility of the systemic and renal alterations induced by Nω-nitro-l-arginine-methyl ester (l-NAME) was assessed by treatment with irbesartan and enalapril (30 and 10 mg/kg per 24 h, respectively) alone or in combination. Design l-NAME (20 mg/kg per 24 h) was given to rats for 6 weeks and treatments were administered during the last 2 weeks. Glomerular filtration rate and renal plasma flow [GFR and RPF per g of kidney weight (KW)] were determined using the clearance technique. Results Arterial pressure was similarly reduced by treatments. GFR was lower in l-NAME-treated rats than in controls (552 ± 52 versus 1106 ± 78 μl/min per g KW), whereas RPF was reduced to a larger extent, thus resulting in an increase in filtration fraction. GFR was normalized by irbesartan but not enalapril or the combination (1042 ± 50, 790 ± 79 and 725 ± 38 μl/min per g KW, respectively). RPF returned to normal and filtration fraction fell markedly with the combination. All treatments reduced the lesions of preglomerular vessels and reversed l-NAME-induced albuminuria and cardiovascular hypertrophy. At a dose of 3 mg/kg per 24 h, irbesartan only reduced the lesions of the afferent arteriole. Conclusions Through its effects on AT1 receptors, angiotensin II may play an important role in the maintenance of l-NAME hypertension and associated alterations. The lower GFR and filtration fraction observed with enalapril in the presence of irbesartan suggests the intervention of non-angiotensin II-mediated mechanism, and at the postglomerular level, in the effect of angiotensin-converting enzyme (ACE) inhibitors.

Evidence for a postsynaptic effect of captopril in isolated perfused rabbit kidney

The influence of captopril (SQ 14225) on the vascular and norepinephrine-releasing responses to nerve stimulation (2, 5 and 10 Hz) was assessed in isolated blood-free perfused rabbit kidney. At a concentration of 0.23 and 0.46 mM in the perfusion medium, captopril markedly attenuated the vasoconstrictor response but did not influence the release of norepinephrine produced by nerve stimulation. These results suggest that captopril may act as an alpha-antagonist at a postjunctional level.

Renal function and structure in a rat model of arterial calcification and increased pulse pressure

Clinical studies suggest a strong link between tissue calcification and pressure hyperpulsatility in end stage renal disease patients. Using a Wistar rat model of arterial elastocalcinosis and hyperpulsatility [vitamin D and nicotine (VDN) treatment], we evaluated the relative importance of tissue calcification and hyperpulsatility in the etiology of renal failure. VDN rats showed significant increases in aortic wall calcium content (50 times; 992+/-171 vs. control 19+/-1 micromol/g dry wt) and pulse pressure (1.5 times; 61+/-4 vs. control 40+/-2 mmHg). Significant renal calcification (16 times; 124+/-27 vs. control 8.1+/-0.7 micromol/g dry wt) occurred mainly within the media of the preglomerular vasculature and in the areas of interstitial fibrosis in VDN. Extensive renal damages (5 times; 26+/-5% of collapsed-atrophic or sclerotic glomeruli, or glomerular cysts vs. control 5.2+/-0.3%; 28 times; 61+/-12% areas of focal, cortical areas exhibiting interstitial fibrosis per section vs. control 2.2+/-0.6%) were observed histologically. The glomerular filtration rate significantly decreased (880+/-40 vs. control 1,058+/-44 microl.min(-1).g kidney wt(-1)). Albuminuria increased six times (1.6+/-0.4 vs. control 0.27+/-0.04 mg/24 h). There were significant linear relationships between albuminuria and pulse pressure (r2=0.408; n=24) or renal calcium content (r2=0.328; n=24; P<0.05) and between glomerular filtration rate and pulse pressure (r2=0.168; n=27). To our knowledge, this study provides the first evidence of links between both 1) hyperpulsatility and renal dysfunction, and 2) renal calcification and renal dysfunction. Given the increasing frequency of end-stage renal disease, this model could prove useful for preclinical evaluation of drugs that prevent or attenuate hyperpulsatility and/or tissue calcification.

New Method for Imaging Innervation of the Renal Preglomerular Vasculature. Alterations in Hypertensive Rats

Objective: To develop a new method for viewing adrenergic innervation along renal preglomerular vessels; to assess nerve densities and vascular lesions along arcuate arteries (ArcA), arcuate arterial branches (ArcB), and interlobular arteries (ILA) in spontaneously hypertensive rats (SHR) and in angiotensin II (AngII) and in NG‐nitro‐l‐arginine methyl ester (l‐NAME) hypertensive rats.

IGF-I Enhances Cellular Proliferation and Apoptosis in Growing Rats with Progressive Chronic Renal Failure † 1790

IGF-I Enhances Cellular Proliferation and Apoptosis in Growing Rats with Progressive Chronic Renal Failure † 1790