Renal Microvessels Research Articles

2′,3′-cAMP, 3′-AMP, and 2′-AMP inhibit human aortic and coronary vascular smooth muscle cell proliferation via A2B receptors

Rat vascular smooth muscle cells (VSMCs) from renal microvessels metabolize 2',3'-cAMP to 2'-AMP and 3'-AMP, and these AMPs are converted to adenosine that inhibits microvascular VSMC proliferation via A(2B) receptors. The goal of this study was to test whether this mechanism also exists in VSMCs from conduit arteries and whether it is similarly expressed in human vs. rat VSMCs. Incubation of rat and human aortic VSMCs with 2',3'-cAMP concentration-dependently increased levels of 2'-AMP and 3'-AMP in the medium, with a similar absolute increase in 2'-AMP vs. 3'-AMP. In contrast, in human coronary VSMCs, 2',3'-cAMP increased 2'-AMP levels yet had little effect on 3'-AMP levels. In all cell types, 2',3'-cAMP increased levels of adenosine, but not 5'-AMP, and 2',3'-AMP inhibited cell proliferation. Antagonism of A(2B) receptors (MRS-1754), but not A(1) (1,3-dipropyl-8-cyclopentylxanthine), A(2A) (SCH-58261), or A(3) (VUF-5574) receptors, attenuated the antiproliferative effects of 2',3'-cAMP. In all cell types, 2'-AMP, 3'-AMP, and 5'-AMP increased adenosine levels, and inhibition of ecto-5'-nucleotidase blocked this effect of 5'-AMP but not that of 2'-AMP nor 3'-AMP. Also, 2'-AMP, 3'-AMP, and 5'-AMP, like 2',3'-cAMP, exerted antiproliferative effects that were abolished by antagonism of A(2B) receptors with MRS-1754. In conclusion, VSMCs from conduit arteries metabolize 2',3'-cAMP to AMPs, which are metabolized to adenosine. In rat and human aortic VSMCs, both 2'-AMP and 3'-AMP are involved in this process, whereas, in human coronary VSMCs, 2',3'-cAMP is mainly converted to 2'-AMP. Because adenosine inhibits VSMC proliferation via A(2B) receptors, local vascular production of 2',3'-cAMP may protect conduit arteries from atherosclerosis.

Microbubble-based synchrotron radiation phase contrast imaging: basic study and angiography applications

The purpose of the study was to evaluate the feasibility of microbubbles as phase contrast imaging (PCI) agents for angiography applications. The hypothesis was that the introduction of microbubbles into tissue produces a significant change in the refractive index and highlights the lumen of the vessel in PCI. The absorption and phase contrast images of commercially available microbubbles were obtained and compared in vitro. A further increase in contrast was observed in PCI. Microbubbles highlighted the lumen of the renal microvessels, acting as a positive contrast medium in ex vivo imaging. In addition, home-made microbubbles with larger diameters were introduced for image contrast enhancement in living tumor-bearing mice, demonstrating the feasibility of microbubble-based x-ray phase-contrast imaging for tumor vasculature in vivo.

Revascularization of swine renal artery stenosis improves renal function but not the changes in vascular structure

Renal revascularization by percutaneous transluminal angioplasty improves blood pressure and stenotic kidney function in selected groups of patients, but the reversibility of intrarenal and microvascular remodeling remains unknown. Here, we tested the hypothesis that renal angioplasty improves the function and structure of renal microcirculation in experimental chronic renal artery stenosis. Stenotic kidney function, hemodynamics, and endothelial function were assessed in vivo in pigs after 10 weeks of unilateral renal artery stenosis. Renal microvascular remodeling, angiogenic pathways, and fibrosis were measured ex vivo. Angioplasty and stenting carried out 4 weeks before measurement decreased blood pressure, improved glomerular filtration rate, and improved microvascular endothelial function. It also promoted the expression of angiogenic factors and decreased renal apoptosis due to stenosis, compared with a sham intervention. The spatial density of renal microvessels, however, was partially improved after angioplasty. Renal blood flow was incompletely restored compared with the kidneys of sham-treated animals, as was interstitial fibrosis. Renal microvascular media-to-lumen ratio remained unchanged by angioplasty. Thus, our study shows that revascularization of a stenotic renal artery restores the glomerular filtration rate and renal endothelial function 4 weeks later. Renal hemodynamics and structure, however, are incompletely resolved.

Impaired endothelial proliferation and mesenchymal transition contribute to vascular rarefaction following acute kidney injury.

Acute kidney injury induces the loss of renal microvessels, but the fate of endothelial cells and the mechanism of potential vascular endothelial growth factor (VEGF)-mediated protection is unknown. Cumulative cell proliferation was analyzed in the kidney of Sprague-Dawley rats following ischemia-reperfusion (I/R) injury by repetitive administration of BrdU (twice daily) and colocalization in endothelial cells with CD31 or cablin. Proliferating endothelial cells were undetectable for up to 2 days following I/R and accounted for only ∼1% of BrdU-positive cells after 7 days. VEGF-121 preserved vascular loss following I/R but did not affect proliferation of endothelial, perivascular cells or tubular cells. Endothelial mesenchymal transition states were identified by localizing endothelial markers (CD31, cablin, or infused tomato lectin) with the fibroblast marker S100A4. Such structures were prominent within 6 h and sustained for at least 7 days following I/R. A Tie-2-cre transgenic crossed with a yellow fluorescent protein (YFP) reporter mouse was used to trace the fate of endothelial cells and demonstrated interstititial expansion of YFP-positive cells colocalizing with S100A4 and smooth muscle actin following I/R. The interstitial expansion of YFP cells was attenuated by VEGF-121. Multiphoton imaging of transgenic mice revealed the alteration of YFP-positive vascular cells associated with blood vessels characterized by limited perfusion in vivo. Taken together, these data indicate that vascular dropout post-AKI results from endothelial phenotypic transition combined with an impaired regenerative capacity, which may contribute to progressive chronic kidney disease.

T-Type Ca Channel Blockade as a Determinant of Kidney Protection

Voltage-dependent Ca channels are classified into several subtypes based on the isoform of their α1 subunits. Traditional Ca channels blockers (CCBs), including nifedipine and amlodipine, act predominantly on L-type Ca channels, whereas novel CCBs such as efonidipine, benidipine and azelnidipine inhibit both L-type and T-type Ca channels. Furthermore, cilnidipine blocks L-type and N-type Ca channels. These CCBs exert divergent actions on renal microvessels. L-type CCBs preferentially dilate afferent arterioles, whereas both L-/T-type and L-/N-type CCBs potently dilate afferent and efferent arterioles. The distinct actions of CCBs on the renal microcirculation are reflected by changes in glomerular capillary pressure and subsequent renal injury: L-type CCBs favor an increase in glomerular capillary pressure, whereas L-/T-type and L-/N-type CCBs alleviate glomerular hypertension. The renal protective action of L-/T-type CCBs is also mediated by non-hemodynamic mechanisms, i.e., inhibition of the inflammatory process and inhibition of Rho kinase and aldosterone secretion. Finally, a growing body of evidence indicates that T-type CCBs offer more beneficial action on proteinuria and renal survival rate than L-type CCBs in patients with chronic kidney disease (CKD). Similarly, in CKD patients treated with renin-angiotensin blockers, add-on therapy with N-type CCBs is more potent in reducing proteinuria than that with L-type CCBs, although no difference is found in the subgroup with diabetic nephropathy. Thus, the strategy for hypertension treatment with CCBs has entered a new era: treatment selection depends not only on blood pressure control but also on the subtypes of CCBs.

Role of renal microcirculation in experimental renovascular disease

Renal artery stenosis (RAS) causes renal injury partly via microvascular (MV) endothelial dysfunction and damage. Vascular endothelial growth factor (VEGF) is crucial for preservation of microvasculature and promotes vascular proliferation and endothelial repair. We have previously shown that MV rarefaction is associated with decreased VEGF in the kidney exposed to chronic RAS, accompanied by deteriorated renal function and fibrosis. We hypothesized that preserving the renal microcirculation in the stenotic kidney will halt the progression of renal damage. Unilateral RAS was induced in 16 pigs. In eight, VEGF (0.05 micrograms/kg) was infused intra-renally at the onset of RAS. After 6 weeks, single-kidney haemodynamics and function were assessed using in vivo multi-detector computed tomography (CT). Renal microvessels, angiogenic pathways and morphology were investigated ex vivo using micro-CT, real-time PCR and histology. Blood pressure and degree of RAS was similar in RAS and RAS + VEGF pigs. Single-kidney renal blood flow (RBF) and glomerular filtration rate (GFR) were reduced in RAS compared to Normal (221.1 +/- 46.5 and 29.9 +/- 3.8 vs. 522.5 +/- 60.9 and 49.3 +/- 3.4 mL/min, respectively, P < 0.05), accompanied by decreased cortical MV density and increased renal fibrosis. Pre-emptive administration of VEGF preserved MV architecture, attenuated fibrosis and normalized RBF and GFR (510.8 +/- 50.9 and 39.9.1 +/- 4.1 mL/min, P = not significant vs. Normal). This study underscores the importance of the renal microcirculation in renovascular disease. Intra-renal administration of VEGF preserved renal MV architecture and function of the stenotic kidney, which in turn preserved renal haemodynamics and function and decreased renal fibrosis. These observations suggest that preventing renal MV loss may be a potential target for therapeutic approaches for patients with chronic renovascular disease.

Intrarenal Artery Delineation With Ultra High Resolution, Flat Panel Based, Volume Computerized Tomography: Outer Limits of Spatial Resolution

New methods of noninvasive high resolution imaging may improve the delineation of tumor microvessels and, thus, be of significant help in surgical planning and cost-effective monitoring of novel anti-angiogenic therapy. We determined the maximum delineation of intrarenal microvessels with a novel flat panel based volume computerized tomography system in an experimental setting. We prospectively evaluated 13 porcine renal specimens for intrarenal vessel delineation using a prototype gantry based, flat panel, cone beam computerized tomography system. The gantry incorporates an array of a 40 x 30 cm(2) CsI amorphous silicon flat panel detector consisting of a 2,048 x 1,536 matrix. After catheterizing the renal artery with a 5Fr end hole catheter a contrast enhanced scan was performed using BaS as contrast medium at a dilution of 200 mg/ml. The diameter of all definable arterial branches was determined using a software tool based on Medical Imaging and Interaction Toolkit, allowing semi-automatic segmentation of the vessel tree. In step 1 the vessel tree is segmented by a 3-dimensional region growing algorithm. Following its medial axis the vessel tree is extracted and converted to a representation, including the diameter of the vessels. In each kidney an average +/- SD of 47,454 +/- 22,382 arterial branches could be delineated. The diameter of the branches was 0.029 (mean 0.032 +/- 0.0025) to 3.444 mm (mean 1.813 +/- 0.6139) with a median of 0.263 mm. Of visible intrarenal arteries 2.7% had a vessel diameter of 0.029 mm. Flat panel based volume computerized tomography can visualize intrarenal microvessels down to a diameter of 0.03 mm. It may improve the assessment of renal microvessel architecture in healthy patients and in those with pathological conditions.

Protective effect of catechin on renal microvessels in 5/6 nephrectomized rats and its mechanism

To investigate the effects of catechin on angiotensin-converting enzyme (ACE) activity, angiotensin II(Ang II) content and microvessel density (MVD) in renal tissues of 5/6 nephrectomized rats. Sixty SD rats were divided into 3 groups: sham-operated group, untreated group and catechin group. Animal model was reproduced by 5/6 nephrectomy. After 8- and 12-week administration, rats were sacrificed. Renal MVD was measured by immunohistochemical method with CD34 marking. Activities of ACE in plasma and renal cortices were measured by ultraviolet spectrophotometer, and Ang II contents in plasma and renal cortices were measured by radioimmunoassay. Glomerular sclerosis index (GSI) and tubule interstitial score (TIS) were calculated by semiquantitative integration with hematoxylin and eosin staining and periodic acid-Schiff staining. After 8- and 12-week administration, the GSI and TIS in the catechin group were much less than those in the untreated group (P < 0.05, P < 0.01). MVDs around glomerulus and tubule at the end of the 8th and 12th week in the untreated group and the catechin group were much less than those in the sham-operated group (P < 0.01). The ACE activities and Ang II contents in plasma and renal cortices in the catechin group were much less than those in the untreated group (P < 0.01). By Pearson correlation analysis, we found that MVD had negative correlation with GSI, TIS, Ang II content, and ACE activity (P < 0.01), however, the ACE activity and Ang II content had positive correlation with GSI, TIS (P < 0.01). Catechin can prevent the 5/6 nephrectomized rats from decreasing of MVD and inhibit the progress of glomerular sclerosis and interstitial fibrosis by inhibiting the activity of ACE and reducing the production of Ang II.

Water extracts of cigarette smoke elicit smooth muscle dependent relaxation of rat renal arteries

The mechanisms responsible for cigarette smoking‐induced increases in glomerular filtration rate and microalbuminuria in healthy persons are not known. We hypothesized that they may be related to changes in renal vascular resistance elicited by the water‐soluble components of cigarette smoke (wCS). The acute effects of wCS on isometric tension of preconstricted isolated rat renal arteries were examined. Increasing doses of wCS (1‐5‐10% of saturated solution) elicited dose dependent relaxations of renal arteries (wCS‐1%: 41±15%, wCS‐5%: 79±9%, wCS‐10% 90±6%). Endothelium removal did not affect relaxations to wCS (wCS‐1%: 15 ± 9%, wCS‐5%: 62±15%, wCS‐10% 90±7%). Similar results were obtained with nicotine free cigarettes (wCS‐1%: 38 ± 3%, wCS‐5%: 77±15%, wCS‐10% 97±0,2%). Relaxation induced by wCS were not affected by inhibition of soluble guanylate cyclase by ODQ (wCS‐1%: 13 ± 4%, wCS‐5%: 58±15%, CS‐10% 94±4%). Thus water soluble components of cigarette smoke elicit relaxations of isolated renal arteries that are independent of endothelium‐, nicotine‐ and cGMP‐related vasomotor mechanisms (such as NO or CO). Although further studies are necessary to elucidate the mechanisms responsible for renal artery relaxation, we propose that cigarette smoking, by modulating directly the smooth muscle tone of renal microvessels, could increases glomerular filtration rate and microalbuminuria observed in healthy smoking men.(Grants: Hungarian SRF/OTKA T48376, K71591, F68638; HSC/ETT 364/2006, AHA F. Aff. 0855910D, USA)

VEGF is a mediator of the renoprotective effects of multipotent marrow stromal cells in acute kidney injury

Adult stem cell treatment of complex disorders is a promising therapeutic approach and multipotent marrow stromal cells (MSCs) have been shown to be effective in various animal models of diseases. Acute kidney injury (AKI) is a common and serious problem in hospitalized patients and bone marrow derived multipotent MSCs have been shown to be effective in different models of AKI. The mechanism of action of MSCs is complex but involves paracrine actions including growth factor secretion. Knockdown of vascular enthothelial growth factor (VEGF) by siRNA reduced effectiveness of MSCs in the treatment of ischemic AKI in a rat model. Animals treated with MSCs had increased renal microvessel density compared to VEGF knockdown MSC-treated and vehicle-treated animals. These results show that VEGF is an important mediator of the early and late phase of renoprotective action after AKI in the context of stem cell treatment.

Colfibrate attenuates blood pressure and sodium retention in DOCA-salt hypertension

Clofibrate, a peroxisome proliferator-activated receptor-alpha (PPAR alpha) agonist, increases renal tubular cytochrome P450 4a (Cyp4a) expression thereby increasing 20-hydroxyeicosatetraenoic acid (20-HETE) production. To determine if clofibrate affects blood pressure regulation we studied mice with DOCA-salt induced hypertension in wild-type and PPAR alpha knockout mice. Wild-type mice treated with DOCA-salt had higher mean arterial pressures and higher cumulative sodium balance, but lower renal 20-HETE production than did vehicle-treated mice. Treating DOCA-salt mice with clofibrate attenuated the increase in mean arterial pressure and cumulative sodium balance while increasing 20-HETE production and renal Cyp4a expression. In contrast the PPAR alpha knockout mice treated with clofibrate and DOCA-salt showed no attenuation in the increase of blood pressure, cumulative sodium balance, renal 20-HETE production or Cyp4a protein expression. Expression of the PPAR alpha protein was greater in proximal tubules than in renal microvessels. Our results show that PPAR alpha pathway induces renal tubular 20-HETE production which affects sodium retention and blood pressure regulation in DOCA-salt-treated mice.

VEGF-121 preserves renal microvessel structure and ameliorates secondary renal disease following acute kidney injury

Acute kidney injury induced by renal ischemia-reperfusion (I/R) compromises microvascular density and predisposes to chronic kidney disease (CKD) and sodium-dependent hypertension. VEGF-121 was administered to rats fed a standard (0.4%) sodium diet at various times following recovery from I/R injury for up to 35 days. VEGF-121 had no effect on the initial loss of renal function, as indicated by serum creatinine levels measured 24 h after injury. Serum creatinine levels declined thereafter, indicative of renal repair. Rats were then switched to an elevated (4.0%) sodium diet for an additional 28 days to induce CKD. The 4.0% sodium diet enhanced renal hypertrophy, interstitial volume, albuminuria, and cardiac hypertrophy relative to postischemic animals maintained on the 0.4% sodium diet. Administration of VEGF-121 from day 0 to 14, day 0 to 35, or day 3 to 35 after I/R suppressed the effects of sodium diet on CKD development, while delayed administration of VEGF-121 from day 21 to 35 had no effect. Endothelial nitric oxide synthase protein levels were upregulated in postischemic animals, and this effect was significantly increased by the 4.0% sodium diet but was not influenced by prior treatment with VEGF. Conversely, microvascular density was preserved in postischemic animals treated with VEGF-121 relative to vehicle-treated postischemic animals. These data suggest that early, but not delayed, treatment with VEGF-121 can preserve vascular structure after ischemia and influence chronic renal function in response to elevated sodium intake.

Identification of Novel Endogenous Cytochrome P450 Arachidonate Metabolites with High Affinity for Cannabinoid Receptors

Arachidonic acid is an essential constituent of cell membranes that is esterified to the sn-2-position of glycerophospholipids and is released from selected lipid pools by phospholipase cleavage. The released arachidonic acid can be metabolized by three enzymatic pathways: the cyclooxygenase pathway forming prostaglandins and thromboxanes, the lipoxygenase pathway generating leukotrienes and lipoxins, and the cytochrome P450 (cP450) pathway producing epoxyeicosatrienoic acids and hydroxyeicosatetraenoic acids. The present study describes a novel group of cP450 epoxygenase-dependent metabolites of arachidonic acid, termed 2-epoxyeicosatrienoylglycerols (2-EG), including two regioisomers, 2-(11,12-epoxyeicosatrienoyl)glycerol (2-11,12-EG) and 2-(14,15-epoxyeicosatrienoyl)glycerol (2-14,15-EG), which are both produced in the kidney and spleen, whereas 2-11,12-EG is also detected in the brain. Both 2-11,12-EG and 2-14,15-EG activated the two cannabinoid (CB) receptor subtypes, CB1 and CB2, with high affinity and elicited biological responses in cultured cells expressing CB receptors and in intact animals. In contrast, the parental arachidonic acid and epoxyeicosatrienoic acids failed to activate CB1 or CB2 receptors. Thus, these cP450 epoxygenase-dependent metabolites are a novel class of endogenously produced, biologically active lipid mediators with the characteristics of endocannabinoids. This is the first evidence of a cytochrome P450-dependent arachidonate metabolite that can activate G-protein-coupled cell membrane receptors and suggests a functional link between the cytochrome P450 enzyme system and the endocannabinoid system.

Effect of iron oxide perfusion on renal micro vessel morphology, and dilatory function and endothelial mRNA expression of mouse aorta

Renal microvessels can be selectively isolated using a combined iron oxide‐sieve technique. Here we investigate a potential damaging effect of the iron oxide perfusion on microvessel morphology and endothelial cells. C57Bl6 mice were perfused with PBS (1% albumin) or PBS (1% albumin) with 1% iron oxide. Histology of the kidney was investigated using thin and hemi‐thin sections. Endothelium mediated vasodilatation was tested in rings of abdominal aorta under isometric conditions. Relative mRNA content of endothelial nitric oxide synthase (eNOS), endothelin 1 (ET‐1), and the platelet endothelial cell adhesion molecule‐1 (PECAM‐1) was determined using quantitative PCR analysis in aorta. The histological investigation did not show damage of microvessels in iron oxide perfused kidneys. There was no difference in the dilatation to acetylcholine in aortas of both groups. The quantitative PCR revealed no differences in the expression of eNOS and ET‐1 comparing iron oxide perfused and PBS perfused vessels. PECAM‐1 expression was enhanced in iron oxide perfused vessels. The study shows that iron oxide perfusion may have a slight effect on the analysis of endothelial mRNA expression as indicated by significant changes in PECAM‐1 expression. Similar dilatory function of aortic rings and similar expression of eNOS and ET‐1 mRNA suggest integrity of endothelial cell layer after iron oxide perfusion in aorta.

Lead exposure effect on angiotensin II renal vasoconstriction

Low levels of chronic lead exposure can produce hypertension and endothelial dysfunction, which could be associated with oxidative stress, changes in vascular tone and an imbalance of endothelial-derived vasoconstriction and vasodilator factors. The aim was to investigate the effect of chronic lead-exposure on angiotensin II-induced vasoconstriction in isolated perfused kidney and microvessels. Male Wistar rats (230-250 g) were treated for 12 weeks with lead acetate (100 ppm, Pbgroup) or pure water (control group). We evaluated the vascular reactivity in the kidneys and renal microvessels in the presence and absence of N(omega)-nitro-L-arginine methyl ester (L-NAME) in both groups. The nitrite concentration in renal perfusate was measured as an index of NO released, renal abundance of 3-nitrotyrosine was measured as well as endothelial NO synthase (eNOS) expression. Oxidative stress was measured by using the oxidative fluorescence dye dihydroethidium (DHE) to evaluate in situ production of superoxide and identified by confocal microscopy. Lead-exposure significantly increased blood pressure, eNOS protein expression, oxidative stress and vascular reactivity to angiotensin II. L-NAME potentiated vascular response to angiotensin II in control group but had no effect on the Pb-group. Nitrites released from the kidney of lead-group was lower compared to the control group while 3-nitrotyrosine was higher. This data suggest that lead-induced hypertension could be caused partially by an altered NOsystem.

Paracrine stimulation of vascular smooth muscle proliferation by diadenosine polyphosphates released from proximal tubule epithelial cells

The purinergic receptor system plays an important role in the regulation of both vascular and tubular functions within the kidney; however, the release of purinergic agonists other than ATP by renal tissue is not known. In this investigation, we determine if kidney tissue is a source of diadenosine polyphosphates, which have high affinity for the P(2X) and P(2Y) receptors. Both diadenosine pentaphosphate and hexaphosphate were identified by matrix-assisted laser desorption ionization-mass spectrometry in extracts purified from both whole porcine kidney and from cloned cells of the LLC-PK1 cell line. Both polyphosphates in nanomolar concentrations were found to significantly stimulate the proliferation of vascular smooth muscle cells derived from rat thoracic aortas. The purinergic-receptor antagonist, suramin, did not significantly affect the growth-stimulatory properties of the polyphosphates. The growth stimulation of vascular smooth muscle cells by platelet-derived growth factor was potentiated by both diadenosine polyphosphates. We conclude that diadenosine polyphosphates are endogenous purinergic agonists of the kidney that have physiologic and pathophysiologic relevance. These epithelial cell metabolic products have vasoregulatory properties while linking the energy supply and tubular function.

Attenuated vasoconstrictor responses to endothelin in afferent arterioles during a high-salt diet.

Endothelin-1 (ET-1) is increased in rats on a high-salt (HS) diet and participates in salt-dependent hypertension. Afferent arterioles (AA) are important for long-term blood pressure control, and therefore we hypothesized that a HS diet would alter their responsiveness to ET-1. Sprague-Dawley rats were fed either a normal-salt (NS; 0.66% NaCl) or HS (8%) diet for 1 wk. Diameters of AA were determined in response to increasing concentrations of big ET-1, ET-1, sarafotoxin 6c (S6c), or norepinephrine (NE), using the blood-perfused juxtamedullary nephron technique. ET-1 responses were also determined during blockade of endothelin type A (ET(A)) or type B (ET(B)) receptors with 10 nM ABT-627 or 30 nM A-192621, respectively. Expression of ET(A) and ET(B) receptors was determined in renal microvessels. Responses of AA to big ET-1, ET-1, and S6c were significantly attenuated during a HS diet (e.g., response to 10(-10) M ET-1 in NS vs. HS rats: -52.5 +/- 10.2 vs. +5.6 +/- 11.3% of control diameter; P < 0.05), with no change in the responses to NE. ET(B), but not ET(A) receptor blockade abolished the different response to ET-1 between the NS and HS groups. ET(B) receptor expression in renal microvessels was increased in response to HS (17.7 +/- 2.4 vs. 6.6 +/- 3.0% of beta-actin, P = 0.02), whereas ET(A) receptor expression was unchanged. These results suggest that the reduced vasoconstrictor response of AA to endothelin peptides during a HS diet is mediated by increased vasodilatory function of endothelial ET(B) receptors. By preserving renal blood flow, this may be an important mechanism to restore sodium balance during a HS diet.

Epoxyeicosatrienoic Acid Inhibition Alters Renal Hemodynamics During Pregnancy

In this study we examined the expression of cytochrome P450 (CYP) 2C and CYP2J isoforms in renal proximal tubules and microvessels isolated from rats at different stages of pregnancy. We also selectively inhibited epoxyeicosatrienoic acid (EET) production by the administration of N-methanesulfonyl-6-(2-proparyloxyphenyl)hexanamide (MSPPOH 20 mg/kg/day iv) to rats during Days 14-17 of gestation and to age-matched virgin rats and determined the consequent effects on renal function. Western blot analysis showed that CYP2C11, CYP2C23, and CYP2J2 expression was significantly increased in the renal microvessels of pregnant rats on Day 12 of gestation. In the proximal tubules, CYP2C23 expression was significantly increased throughout pregnancy, while the expression of CYP2C11 was increased in early and late pregnancy and the expression of CYP2J2 was increased in middle and late pregnancy. MSPPOH treatment significantly increased pregnant rats' mean arterial pressure, renal vascular resistance, and sodium balance but significantly decreased renal blood flow, glomerular filtration rate, and urinary sodium excretion, as well as fetal pups' body weight and length. In contrast, MSPPOH treatment had no effect on renal hemodynamics or urinary sodium excretion in age-matched virgin rats. In pregnant rats, MSPPOH treatment also caused selective inhibition of renal cortical EET production and significantly decreased the expression of CYP2C11, CYP2C23, and CYP2J2 in the renal cortex, renal microvessels, and proximal tubules. These results suggest that upregulation of renal vascular and tubular EETs contributes to the control of blood pressure and renal function during pregnancy.

Endothelin-A Receptor Blockade Improves Renal Microvascular Architecture and Function in Experimental Hypercholesterolemia

Hypercholesterolemia (HC) may trigger early renal injury, partly by impairing the function or the structure of renal microvessels (MV). The endothelin (ET) system is upregulated in HC and can have an impact on the renal microcirculation by regulating MV tone, growth factors, and remodeling. It was hypothesized that ET-A blockade would protect the HC kidney by improving the function and attenuating the damage of intrarenal MV. Single-kidney function and hemodynamic responses to endothelium-dependent challenge were assessed in pigs after 12 wk of experimental HC, HC and chronic supplementation with the ET receptor A blocker ABT-627 (HC+ET-A, 0.75 mg/kg per d), and normal controls. Renal MV architecture then was studied ex vivo using three-dimensional microcomputed tomography imaging, and growth factors and remodeling pathways were explored in renal tissue. The HC kidney showed increased MV density compared with normal (77.68 +/- 5.1 versus 62.9 +/- 4.8 vessels/cm(2); P = 0.04) but blunted endothelial function. Chronic ET-A blockade in HC upregulated renal vascular growth factors, further increased renal MV density (139.9 +/- 8.4 vessels/cm(2); P = 0.001 versus normal and HC), and decreased renal tissue and MV remodeling. Furthermore, ET-A blockade in HC decreased MV tortuosity and improved MV endothelial function, suggesting accelerated stabilization and maturation of neo-vessels. Modulation of renal MV architecture and function in HC is mediated partly by the endogenous ET system. Notably, ET-A blockade enhanced the proliferation and facilitated the maturation of renal MV in the HC kidney and improved renal MV remodeling and function. This study suggests novel renoprotective effects of ET-A blockers and supports further exploration of strategies that target the ET pathway in HC and atherosclerosis.

Epoxyeicosatrienoic Acid Inhibition Alters Renal Hemodynamics During Pregnancy

In this study we examined the expression of arachidonic acid epoxygenases (CYP2C and 2J)in different renal tissues isolated from pregnant rats. We also selectively inhibited EET production by administering N-methanesulfonyl-6-(2-proparyloxyphenyl)hexanamide (MSPPOH, 20 mg/kg/day, iv) to pregnant rats during days 14–17 of gestation and to age-matched virgin rats, and determined the effects on renal function. Western blot analysis showed significant increases in the expression of CYP2C11 (97%), CYP2C23 (21%), and CYP2J2 (164%) in the renal microvessels of Day 12 pregnant rats, as well as that CYP2C23 expression was increased in the proximal tubules throughout pregnancy. MSPPOH-treated pregnant rats had higher mean arterial pressure (MAP) (117 ± 3 versus 97 ± 2 mmHg, P<0.05, n=6) and renal vascular resistance (RVR) (21 ± 1 versus 16 ± 1 mmHg mL -1 min-1, P<0.05, n=6), but lower renal blood flow (RBF) (3.8 ± 0.1 versus 5.9 ± 0.3 mL min-1, P<0.05, n=6) and glomerular filtration rate (GFR) (1.6 ± 0.05 versus 1.9 ± 0.08 mL min-1, P<0.05, n=6). In contrast, MSPPOH treatment had no effect on renal hemodynamics in virgin rats. In pregnant rats, MSPPOH treatment caused selective inhibition of renal cortical EET production and decreased the expression of epoxygenases in the renal cortex, renal microvessels, and proximal tubules. Conclusion: This study demonstrates that CYP epxoygenases are altered in renal microvessels and proximal tubules during pregnancy in rats. Our findings demonstrate that the selective inhibition of EET production causes abnormal renal hemodynamics, and suggest that upregulation of renal vascular and tubular EETs contributes to the control of blood pressure and renal function during pregnancy.