Tail-cuff Systolic Blood Pressure Research Articles

3 Naturally Occurring Perinatal Growth Restriction Prevents the Development of Diet Induced Metabolic Syndrome in Mice.

Background: Epidemiological studies show low birth weight increases the risk of developing metabolic syndrome (obesity, glucose intolerance and hypertension). Identification of an analogous model within an inbred mouse strain would facilitate mechanistic understanding. Hypotheses: 1) Perinatal growth restricted (PGR) mice have altered body composition, impaired glucose tolerance and hypertension (compared to isogenic controls). 2) The effects of PGR are amplified follow provision of a high fat diet. Methods: PGR C57Bl6 pups were identified at weaning (3wk) and paired with same sex normally grown littermates (n = 8 per group). Growth and feed intake was recorded weekly for 34 wk (17 wk on standard diet, then 17 wk on high fat diet). Glucose tolerance tests (GTT) and tail cuff systolic blood pressure (SBP) measurements were performed at the end of each diet period. Results: PGR pups weighed significantly less than controls throughout the study, despite increased caloric intake (P<0.05). PGR adults gained less than half as much weight as controls while on the high fat diet (HF). While on HF, glucose tolerance was significantly better in PGR mice than controls. Although SBP was significantly higher in PGR mice at 20 wk, following 17 wk of HF, SBP was lower in the PGR mice (P<0.05). Conclusion: Naturally occurring perinatal growth restriction is associated with (1) impaired postnatal weight gain; (2) increased SBP while on standard diet; and (3) decreased propensity towards diet-induced metabolic syndrome while on a high fat diet.

Effect of lisinopril on rat liver tissues in L-NAME induced hypertension model

N(G)-Nitro-L-arginine methyl ester hydrochloride (L-NAME) is a non-specific nitric oxide (NO) inhibitor and it has been used to eliminate the role of NO in many studies like animal models for hypertension. In this study, we aimed to investigate whether lisinopril treatment has any biochemical and/or histopathological effect on rat liver tissue in a L-NAME-induced hypertension model. Forty-eight 6-weeks-old male Spraque-Dawley rats were used in the study. The animals used in the study were randomly divided into four equal groups. To induce hypertension, L-NAME was added to drinking water at a concentration of 600 mg/l and each rat was given 75 mg/kg/day of L-NAME for 6 weeks. Tail cuff systolic blood pressure (SBP) was measured at first, third, and sixth weeks. There was a significant difference between the experiment groups and controls. In only lisinopril given and L-NAME plus lisinopril administered groups, each rat was given 10 mg/kg of lisinopril for 6 weeks. At the end of the study, the animals were sacrificed. Blood and tissue samples were collected for biochemical and histopathological analysis. It has been observed that mean NO level was significantly decreased in L-NAME given group (p<0.05). Mean ALT levels were significantly increased in lisinopril and L-NAME plus lisinopril given groups, when compared with the control group (p<0.05). AST levels were in normal range in all groups (p>0.05). Hepatocyte degeneration was prominent in lisinopril given group, whereas mononuclear cell infiltration was significant in L-NAME given groups. Although the beneficial effects in L-NAME-induced hypertension treatment, lisinopril can lead to some unexpected results like hepatocyte degeneration, serum enzyme level elevation, and slight mononuclear cell infiltration.

Adult Hypertension in Intrauterine Growth-Restricted Offspring of Hyperinsulinemic Rats

In humans, intrauterine growth-restricted newborns are prone to develop hypertension as adults. We studied a rat model of pregnancy-induced hypertension associated with intrauterine growth restriction (IUGR) produced by chronic administration of insulin. Fetuses of hyperinsulinemic dams (HDs) were smaller than those of normal dams (5.1+/-0.4 g versus 5.6+/-0.1 g, respectively; P<0.05). At 16 weeks of age, tail-cuff systolic blood pressure was measured, the rats were placed in metabolic cages and euthanized, and the kidneys were examined. Male but not female offspring of HDs (n=9) had higher blood pressure than normal-pregnancy offspring (n=12; 148+/-11 mm Hg versus 118+/-14 mm Hg; P<0.004). In contrast to other models, there was no difference in ours in the number and volume of glomeruli. However, there were significantly greater glomerular, tubulointerstitial, and vascular damage indices in the kidneys of male HD offspring versus controls (2.01+/-0.34 versus 1.08+/-0.16, 1.80+/-0.34 versus 0.76+/-0.12, and 2.13+/-0.81 versus 0.78+/-0.16, respectively; P<0.0001), with similar tubulointerstitial findings in females. Increased expression of collagen type IV, a kidney damage marker indicating fibrosis, was found in the tubulointerstitium. This may be associated with downregulation of bone morphogenetic protein 6, a presumptive antifibrogenic agent, at the end of gestation. In conclusion, male offspring of HDs displayed IUGR and adult hypertension accompanied by several indices of renal fibrosing damage, mainly in the renal tubulointerstitium. Our findings suggest that there is >1 pathway of fetal programming leading from IUGR to development of hypertension in later life.

Effects of glitazones on blood pressure and vascular structure in mesenteric resistance arteries and basilar artery from genetically hypertensive rats

1. The effects of two of the glitazone (thiazolidinedione) class of drugs, namely rosiglitazone and pioglitazone, on blood pressure and vascular remodelling in the New Zealand genetically hypertensive (GH) rat model were investigated. 2. In the first study, a GH group given rosiglitazone (5 mg/kg per day) from the age of 7 to 12 weeks was compared with a GH control group. In the second study, GH rats were given either pioglitazone, simvastatin, valsartan or combinations of pioglitazone with simvastatin or valsartan (all drugs at a dose of 10 mg/kg per day). 3. Tail-cuff systolic blood pressure was measured weekly. At the end of the experiment, blood vessels were fixed by perfusion and samples of mesenteric resistance arteries (MRA), second-order branches and basilar artery were embedded in Technovit and serial sections were cut and stained with Giemsa for stereological analysis. Media width, medial cross-sectional area and lumen diameter were determined and the ratio of media width/lumen diameter was calculated. 4. Rosiglitazone significantly reduced blood pressure in GH rats. 5. In MRA, rosiglitazone had a hypotrophic effect on media, reduced lumen diameter and reduced media/lumen ratio (P<0.001). 6. In basilar artery, there was also a hypotrophic effect of rosiglitazone on media and reduced media/lumen ratio (P<0.001). 7. Pioglitazone slowed down the rate of blood pressure increase with age in GH rats and had a greater effect on blood pressure when given in combination with simvastatin. 8. Pioglitazone had a hypotrophic effect on the media of MRA and basilar artery. The hypotrophic effect was enhanced when pioglitazone was given in combination with simvastatin. The media/lumen ratio was reduced by pioglitazone; in MRA, combination treatment with simvastatin reduced the ratio further to normal and, with valsartan, to below normal. In basilar artery, the media/lumen ratio was reduced further by both combination treatments, but was lowest in the pioglitazone-valsartan combination group. 9. The significant effects on MRA and basilar artery structure (and, thus, haemodynamics) seen after rosiglitazone monotherapy and after pioglitazone, given alone and in combination with simvastatin or valsartan, may well indicate a glitazone class effect on vascular structure and, hence, cardiovascular function.

Male Gender and Not the Severity of Hypertension Is Associated With End-Organ Damage in Aged Stroke-Prone Spontaneously Hypertensive Rats

It is well-known that gender affects the progression of kidney failure. Male patients exhibit faster development of age-dependent renal disease than do women. In the present study, we examined arterial blood pressure (BP), proteinuria, and end-organ damage in male and female retired breeders from our colony of stroke-prone spontaneously hypertensive rats (SHRSP). Male (n = 7) and female (n = 11) SHRSP littermates maintained on Purina Laboratory Chow 5008 and water were studied starting at 53 weeks of age. Systolic BP was measured by tail-cuff plethysmography and 24-h urinary protein excretion was quantified while animals were housed in metabolic cages. Blood was obtained by retro-orbital bleeding. Mean arterial pressure (MAP) was then monitored by radiotelemetry. Organs were preserved for histopathologic assessment. Tail-cuff systolic BP did not differ between the sexes. Male SHRSP exhibited greater proteinuria (128 +/- 7 mg/d) than females (21 +/- 5 mg/d, P < .001). Blood urea nitrogen was higher in males (22 +/- 2 mg%) v females (15 +/- 1 mg%, P < .005). The MAP by radiotelemetry did not differ between the sexes (179 +/- 3 mm Hg in males v 192 +/- 6 mm Hg in females, 2 weeks after probe implantation). Stroke-related mortality was greater in males (83%) than females (10%). Renal vascular disease including thrombotic microangiopathy affecting glomeruli and microvessels and cardiac damage were more prominent in male SHRSP. These findings demonstrate that male gender is a major risk factor for multisystem end-organ damage associated with aging and hypertension in SHRSP, despite comparable degrees of hypertension among males and females.

Exercise Training Increases Beta Adrenergic Responsiveness In Hypertension

Cardiac responses to beta-adrenergic receptor (BAR) stimulation are depressed in chronic hypertension. We tested whether exercise training (ET) could improve BAR responsiveness in spontaneous hypertensive rats (SHR) and whether BAR kinase (GRK2) plays a role after exercise in SHR. Methods Female, SHR (age: 4mo) were placed into a treadmill running (SHR-TRD, n=12; 20m/min, 1hr/day, 5d/wk, 12 wks) or sedentary group (SHR-SED, n=12). Age-matched WKY rats (n=12) were controls. Tail cuff systolic and diastolic blood pressures were determined. Isovolumic, Langendorff performance was measured (4.5 Hz, LVEDP = 12mmHg, coronary flow=16 ml/min, 2mM Ca2+). Following baseline, 5-minute infusions of isoproterenol (ISO) were administered (10−10 - 10−7 mol/L). Western blot analysis, myocardial morphometry, and histomorphometry were performed. Results Mean blood pressure was higher in SHR vs. WKY (P < 0.05) and unaltered with ET. Myocardial wall thickness and cell surface area were greater in SHR than WKY (P < 0.05) and augmented with exercise training (P < 0.05). At baseline, LV developed pressure (LVDevP) was greater in SHR vs. WKY (WKY: 154 ± 23; SHR-SED: 173 ± 10; SHR-TRD: 175 ± 5 mmHg). The peak response to ISO was shifted rightward 100-fold in both SHR groups relative to WKY. The peak ISO LVDevP response was similar between WKY and SHR-TRD, while the peak response of SHR-SED was blunted (WKY: 38 ± 11%; SHR-SED: 19 ± 9%; SHR-TRD: 49 ± 7%, P < 0.05) (Figure 1A). L type Ca2+ channel (LTCC) and the Na+/Ca2+ exchanger (NCX) abundance were increased in both SHR groups. Training increased ryanodine phosphorylation (RyRp) with no difference in ryanodine receptor density (RyRt) among groups. GRK2 levels were increased in SHR-SED vs. SHR-TRD relative to WKY (SED: 97.65 ± 38%; TRD: 17.53 ± 19%, P < 0.05) (Figure 1B.).Figure 1AFigure 1BConclusion BAR responsiveness is improved with exercise training in SHR, an effect at least in part, secondary to a downregulation of GRK2. Supported by the American Heart Association, Mid-Atlantic Affiliate (JRL), and the National Institute of Health SRH (HL33921) and WJK (HL61690 and HL56205).

Optimizing microarray in experimental hypertension

Genetic noise between outbred animals can potentially be a major confounder in the use of microarray technology for gene expression profiling. The study of paired organs from the same animal offers an alternative approach (e.g., for studies of the kidney in experimental hypertension). The present study was undertaken to determine the level of genetic noise between outbred adult Sprague-Dawley (SD) rats, and to determine the effects of unilateral nephrectomy on changes in gene expression as a basis for the design of microarray studies in experimental hypertension. Male SD rats (approximately 130 g) were acclimatized before measurement of tail-cuff systolic blood pressure (SBP) for 6 control days and 4 days of saline treatment. Left kidney nephrectomy was performed, and the tissue snap-frozen in liquid nitrogen for subsequent RNA extraction. Two weeks later, SBP was measured over 4 control and 8 saline treatment days, and the remaining right kidney removed and frozen. Total RNA purification, preparation of cRNA, hybridization, and scanning of the Rat U34A Affymetrix arrays were performed, and data analyzed using MAS5 software Affymetrix Suite (v5), Bioconductor, as well as statistical methods motivated by relevant simulations. Gene expression profiles in the left control kidney were extremely consistent across animals. The expression profiles of pairs of kidneys from the same animal were, however, more similar than those of kidneys from different animals. Nephrectomy had little effect on the gene expression profiles in the time frame examined. Despite the outbred nature of the rats used in this study, they are useful for gene expression profiling comparisons. The use of paired organs from an individual animal ensures even further genetic identity, allowing determination of genes modified by the treatment of interest.

Stimulation of calcitonin gene-related peptide synthesis and release: mechanisms for a novel antihypertensive drug, rutaecarpine.

Previous investigations have demonstrated that capsaicin-sensitive primary sensory nerves play an important role in modulation of the peripheral resistance of the circulation system. The vanilloid receptor subtype 1 (VR1) is expressed almost exclusively in the primary sensory nerves and cell bodies of these sensory neurons. Rutaecarpine (Rut) can relax vascular smooth muscle via stimulation of calcitonin gene-related peptide (CGRP) release by activation of VR1. In the present study, we examined the depressor effect of Rut and the possible mechanisms in the phenol-induced hypertensive rats, in which hypertension was induced by injecting 50 microl of 10% phenol in the lower pole of the left kidney. Acute administration of Rut (30, 100 or 300 microg/kg, i.v.) caused a depressor effect concomitantly with an increase in the plasma concentration of CGRP in a dose-dependent manner, which was blocked by capsaicin (used to deplete the CGRP from sensory nerves) or capsazepine (a competitive VR1 antagonist), causing an approximately 85% and approximately 80% change in mean arterial pressure, respectively, and by either of them, causing an approximately 90% elevation of plasma CGRP. In the chronic study, Rut at a dose of 3 or 6 mg/kg per day significantly lowered tail-cuff systolic blood pressure to 159 +/- 8 and 136 +/- 10 mmHg, respectively, compared with hypertensive rats (179 +/- 8 mmHg), and caused a sustained hypotensive effect from day 6 on. Pretreatment with capsaicin blocked the depressor effect of Rut by approximately 65%. Treatment with Rut significantly increased the synthesis and release of CGRP, as shown by the increase in the levels of CGRP mRNA and peptide in the dorsal root ganglia, the density of CGRP immunoreactive nerve fibers in the mesenteric artery, the CGRP content in the spinal cord and the plasma concentration of CGRP, which was markedly attenuated by pretreatment with capsaicin. These results suggest, for the first time, that the hypotensive effect of Rut is mediated by stimulation of CGRP synthesis and release via activation of VR1 in the phenol-induced hypertensive rat.

Is there an association between level of adult blood pressure and nephron number or renal filtration surface area?

Reductions in renal filtration surface area (FSA) have been linked to development of hypertension. This study investigated whether there are direct relationships, in the adult rat, between levels of blood pressure and nephron number or total renal FSA. F1 and F2 offspring were generated from a spontaneously hypertensive rat (SHR)/Wistar Kyoto (WKY) rat cross. Tail-cuff systolic blood pressure was measured twice weekly from 5 to 15 weeks of age and mean arterial blood pressure determined prior to sacrifice. At 15 weeks of age, the rats were perfusion-fixed and glomerular (and thereby nephron) number, glomerular size, total length, and surface area of glomerular capillaries and total renal FSA were determined using unbiased stereologic techniques. In F1 offspring, blood pressure levels were midway between the SHR and WKY rats. Nephron number was significantly higher in the WKY rats compared to the SHR and F1 offspring. However, there was no difference in nephron number between the F1 rats and SHR and no difference in renal FSA between the three groups. In the F2 generation, where there is random segregation of the SHR and WKY genes, there was no significant correlation between either nephron number and adult blood pressure (r2= 0.16, P= 0.11) or total renal FSA and adult blood pressure (r2= 0.02, P= 0.58). There was a significant inverse correlation between nephron number and glomerular size (r2= 0.49, P= 0.0043). There is not a direct corollary between nephron number or renal FSA and level of blood pressure in this rat model.

Does a Nephron Deficit in Rats Predispose to Salt-Sensitive Hypertension?

Aim: This study tested the hypothesis that a nephron deficit predisposes rats to salt-sensitive hypertension in adulthood. Methods: Female Wistar-Kyoto rats were fed a low (9%) or a normal (20%) protein diet during pregnancy and lactation. Male, birth-weight-matched offspring were paired. One rat from each pair was perfusion fixed at 4 weeks of age and the other rat at 40 weeks of age. Kidneys were removed and nephron number and total renal filtration surface area (FSA) determined using unbiased stereological techniques. The rats that were allowed to grow to adulthood had tail-cuff systolic blood pressure and body weight determined twice weekly. Between 30 and 40 weeks of age, a normal or a high-salt diet was fed to the rats. Results: The offspring of rats fed the low-protein diet were significantly smaller at birth, and at 4 weeks of age they had a significant reduction in kidney volume, nephron number, and total renal FSA when compared to controls. Tail-cuff systolic blood pressure in the offspring from 4 to 29 weeks of age did not significantly differ between the two groups. Administration of a high-salt diet from 30 to 40 weeks of age led to a significant increase in blood pressure in both dietary treatment groups; however, it was not exacerbated in the rats exposed to the low-protein diet in utero. Conclusions: Maternal protein restriction in rats did not lead to salt-sensitive hypertension. Nephron endowment and FSA did not correlate with blood pressure in adulthood.

Nephron number and blood pressure in rat offspring with maternal high-protein diet.

This study investigated the effects of a high-protein diet during pregnancy on nephron endowment and subsequent levels of blood pressure in the offspring. Female WKY rats were fed either a normal (20%, NPD) or a high (54%, HPD) protein diet during pregnancy. Male offspring were paired at birth. At 4 weeks of age, 1 of the pair was randomly chosen for perfusion fixation, and total glomerular number, and thereby nephron number, was estimated using an unbiased stereological technique. The other rat of the pair was allowed to grow to 30 weeks of age, during which time tail cuff systolic blood pressure was monitored twice weekly. There was no effect of the HPD on birth weight (NPD 4.23+/-0.53 g, HPD 4.26+/-0.45 g, mean+/-SD), kidney weight (NPD 0.372+/-0.049 g, HPD 0.337+/-0.090 g), or total nephron number (NPD 27,191+/-3,512, HPD 26,738+/-4,735). Systolic blood pressure at 30 weeks was 170+/-14 mmHg in NPD and 169+/-14 in HPD offspring. These findings show that a HPD during pregnancy did not lead to an increase in birth weight, kidney weight, or nephron endowment, nor did the HPD affect adult blood pressure.

Fluvastatin remodels resistance arteries in genetically hypertensive rats, even in the absence of any effect on blood pressure.

1. The aims of the present study were, first, to determine whether, in the genetically hypertensive (GH) rat, fluvastatin would lower blood pressure and remodel mesenteric resistance arteries (MRA) and the basilar artery and, second, to see whether treatment with a combination of fluvastatin and the angiotensin receptor antagonist valsartan would have any extra beneficial effect on blood pressure and vascular remodelling. 2. Male GH rats had tail-cuff systolic blood pressure (SBP) monitored weekly from the age of 7 to 12 weeks. Groups (n = 12-14) were treated with fluvastatin (4 mg/kg per day), valsartan (5 mg/kg per day), both mixed in with chow, or a combination of fluvastatin 4 mg/kg per day + valsartan 5 mg/kg per day. Untreated GH and a group of normotensive Wistar (N) rats served as control groups. 3. At 12 weeks of age, intra-arterial (i.a.) blood pressure was measured by femoral cannulation and rats were then perfused (at the SBP of the animal) with Tyrode's solution containing heparin and papaverine followed by 2.5% glutaraldehyde in Tyrode's solution; MRA and basilar arteries were embedded in Technovit. Serial sections were cut and Giemsa stained and stereological methods used to obtain media width, lumen diameter, medial cross-sectional area (CSA) and the ratio of media width to lumen diameter. Hearts were weighed to determine left ventricular (LV) mass. 4. Fluvastatin had no effect on blood pressure or LV mass, whereas valsartan given alone or with fluvastatin significantly reduced both parameters. 5. In MRA, fluvastatin reduced medial CSA, increased lumen size and, therefore, probably decreased vascular resistance. The media/lumen ratio was reduced to a level below that seen with the combination treatment and to below that of the N group. 6. In the basilar artery, fluvastatin and valsartan showed similar outward remodelling of the lumen and reduction in the media/lumen ratio. The combination treatment group showed, in addition, a reduction in medial CSA and an even lower ratio than the GH group on fluvastatin or valsartan alone or the N group. 7. Although fluvastatin has no effect on blood pressure, it does cause significant remodelling of MRA and the basilar artery. These beneficial structural changes in a peripheral resistance artery bed and in an artery involved in regulating resistance in the brain are worthy of further study.

Effect of AT(1) receptor blockade on cardiac apoptosis in angiotensin II-induced hypertension.

Angiotensin II (ANG II) via AT(1) receptors induces apoptosis in cardiomyocytes in vitro. We tested the hypothesis that in vivo AT(1) receptor stimulation is accompanied by cardiac apoptosis and attempted to elucidate the molecular mechanisms involved in the death signaling pathway. Male Sprague-Dawley rats received ANG II (120 ng x kg(-1) x min(-1) sc) for 7 days with or without the AT(1) receptor antagonist losartan (10 mg x kg(-1) x day(-1) orally). Cardiac function was assessed by echocardiography. Apoptosis in the heart was detected and quantified by in situ TdT-mediated dUTP nick-end labeling (TUNEL) and radiolabeled DNA laddering. Expression of bax, bcl-2, caspase 3, and AT(1) and AT(2) receptors was examined by Western blot analysis. Activity of caspase 3 was also measured by a fluorometric immunosorbent enzyme assay. Tail cuff systolic blood pressure was elevated (P < 0.01, n = 6) in ANG II-infused rats (173 +/- 3 mmHg) versus controls (111 +/- 2 mmHg) and reduced by losartan (134 +/- 4 mmHg). Cardiac function was essentially unchanged in ANG II-infused rats. Increased internucleosomal DNA cleavage by TUNEL assay and radiolabeled DNA laddering showed results compatible with enhanced cardiomyocyte apoptosis in the hearts of ANG-II infused rats. The bax-to-bcl-2 ratio, expression of the active form of caspase 3 (17 kDa), and activity of caspase 3 in the hearts of the ANG II group increased more than twofold above controls. Protein expression of AT(1) and AT(2) receptors was significantly increased in ANG II-infused rats compared with control rats. Losartan-treated ANG II-infused rats exhibited normalized apoptosis, bax, caspase 3 activity, and AT(1) receptors. ANG II stimulation of AT(1) receptors in the heart in vivo is associated with an increased rate of apoptosis without major hemodynamic consequences. Bax and caspase 3 are involved in the apoptotic signaling pathway in this experimental paradigm.

Ramipril improves hemodynamic recovery but not microvascular response to ischemia in spontaneously hypertensive rats

Background Angiotensin converting enzyme (ACE) inhibition exerts positive effects on the microvasculature of normotensive animals, although this concept is not universally accepted. Recently, ACE inhibitors have been suggested to be useful for rescue in peripheral ischemia. Methods We investigated whether chronic treatment with the ACE inhibitor ramipril may have a positive impact on the defective healing response to ischemia that is typical of spontaneously hypertensive rats (SHR). Unilateral limb ischemia was induced in 20-week-old SHR by surgically removing the left femoral artery. Rats were allowed to regain consciousness and then were randomly allocated to treatment with ramipril (1 mg/kg body weight in drinking water) or vehicle for 28 days. Results The SHR failed to develop reparative angiogenesis in response to ischemia, thus having inadequate perfusion recovery. Ramipril reduced both tail-cuff systolic blood pressure (180 ± 7 v 207 ± 2 mm Hg in the vehicle group at 28 days, P < .05) and intra-arterial mean blood pressure (115 ± 6 v 135 ± 5 mm Hg in the vehicle group, P < .05). These effects were associated with increased responsiveness to endothelium-dependent vasodilatation by acetylcholine. Treatment with ramipril did not influence muscular capillary and arteriole density but accelerated the rate of perfusion recovery, leading to complete healing within 28 days after surgery. Conclusions These results indicate that ACE inhibition by ramipril may be useful for the treatment of peripheral vascular complications in hypertension.

Normal perivascular sensory dilator nerve function in arteries of Zucker diabetic fatty rats

Background Type II diabetes in humans is associated with pathology of both the cardiovascular and peripheral sensory nervous systems. Because abnormal vasodilator responses have been reported in animals of type II diabetes and perivascular sensory nerves are a source of vasodilator substances, we tested the hypothesis that sensory nerve-dependent relaxation is abnormal in arteries of the Zucker diabetic fatty (ZDF) rat model of type II diabetes. Methods The ZDF rats and genetic controls were studied at 26 weeks of age. Tail-cuff systolic blood pressure (BP) was measured, serum was obtained for chemical determinations, and mesenteric branch arteries were isolated for wire myograph analysis and confocal-based measurement of calcitonin gene-related peptide (CGRP) positive nerve density. Results No differences in BP were detected. Serum glucose, triglycerides, and cholesterol were significantly elevated in ZDF. Sensory nerve-dependent vasodilation was assessed by measuring relaxation of phenylephrine preconstricted arterial segments to cumulative addition of divalent calcium ion (Ca 2+) or capsaicin. Neither Ca 2+- nor capsaicin-induced relaxation were different in ZDF versus control (maximal ZDF response to Ca 2+ = 64% ± 2% v 59% ± 4%; ED 50 for Ca 2+ = 3.7 ± 0.5 mmol/L v 3.2 ± 0.5 mmol/L; n = 5, P = not significant [NS]; maximal ZDF response to capsaicin = 68% ± 9% v 74% ± 4%; ZDF ED 50 = 3.8 ± 0.5 nmol/L v 9.8 ± 7 nmol/L; n = 5, P = NS). In contrast, the maximal relaxation response to acetylcholine was impaired in ZDF (maximal ZDF response = 83% ± 5% v 94% ± 2%, n = 4, P = .039; ED 50 for acetylcholine = 8.1 ± 2.9 nmol/L for ZDF v 33.5 ± 18.2; n = 4 per group, P = .086). The CGRP positive nerve density was not different between groups. Conclusions Blood pressure, perivascular sensory nerve CGRP content, and dilator function is normal in the ZDF model of type II diabetes, whereas endothelium-dependent relaxation is impaired.

Effect of angiotensin-converting enzyme inhibition on renal filtration surface area in hypertensive rats

Angiotensin-converting enzyme (ACE) inhibitor treatment leads to protective effects on the cellular structure of the glomerulus and the kidney. The aim of this study was to determine whether ACE inhibition increases renal filtration surface area in the spontaneously hypertensive rat (SHR). SHR were treated with the ACE inhibitor perindopril at a high dose (3 mg/kg/day) or a low dose (0.1 mg/kg/day) during the period of hypertension development, from 7 to 14 weeks of age. Some animals were treated concomitantly with the bradykinin B2 receptor antagonist, S16118. Tail-cuff systolic blood pressure and body weights were measured twice weekly. At termination of treatment, glomerular number and volume, length, and surface area of glomerular capillaries and renal filtration surface area were estimated using unbiased stereological techniques. There were significant dose-related reductions in blood pressure with high- and low-dose perindopril treatment. Neither low- nor high-dose perindopril treatment had any effect on glomerular number or size or glomerular capillary length and surface area. Hence, there was no significant difference in total renal filtration surface area between any of the experimental groups (8721 +/- 610 mm2 in untreated SHR and 7879 +/- 338 mm2 and 8767 +/- 437 mm2 in the low and high dose perindopril-treated groups, respectively). Coadministration of the bradykinin antagonist did not affect any of the glomerular parameters. ACE inhibition during the period of hypertension development does not lead to an enhanced glomerular capillary growth or increases in total renal filtration surface area in this model.

Role of renin-angiotensin-aldosterone system in salt-sensitive hypertension induced by sensory denervation.

To define the role of the renin-angiotensin-aldosterone system in a novel salt-sensitive model, neonatal Wistar rats were given capsaicin (50 mg/kg sc) on the first and second days of life. After weaning, male rats were divided into the following six groups and treated for 3 wk with: control + normal sodium diet (CON-NS), CON + high-sodium diet (CON-HS), CON + HS + spironolactone (50 mg x kg(-1) x day(-1), CON-HS-SP), capsaicin pretreatment + NS (CAP-NS), CAP-HS, and CAP-HS-SP. Radioimmunoassay shows that plasma renin activity (PRA) and plasma aldosterone level (PAL) were suppressed by HS, but they were higher in CAP-HS than in CON-HS and CON-HS-SP (P < 0.05). Both tail-cuff systolic blood pressure and mean arterial pressure were higher in CAP-HS than in all other groups (P < 0.05). Urine water and sodium excretion were increased with HS intake, but they were lower in CAP-HS than in CON-HS (P < 0.05). Western blot did not detect differences in adrenal AT1 receptor content. Therefore, insufficiently suppressed PRA and PAL in response to HS intake by sensory denervation may contribute to increased salt sensitivity and account for effectiveness of spironolactone in lowering blood pressure in this model.

Candesartan prevents L-NAME-induced cardio-renal injury in spontaneously hypertensive rats beyond hypotensive effects.

Our goal was to assess the cardiovascular and renal protection afforded by angiotensin II type 1-receptor blockade against NG-nitro-L-arginine methyl ester (L-NAME)-exacerbated hypertension in young spontaneously hypertensive rats (SHR), in comparison with the antihypertensive drug, hydralazine. Male SHR were assigned to four groups (n=8 per group): no treatment (controls); L-NAME-treated group (20 mg/kg/day, 10 days, orally); co-treatment with L-NAME and hydralazine (15 mg/kg/day, by gavage); co-treatment with L-NAME and candesartan cilexetil (10 mg/kg/day, by gavage), i.e. at a dose that inhibited acute pressor responses to 5-20 ng angiotensin II. One animal died in the L-NAME group, and tail-cuff systolic blood pressure (SBP) increased significantly compared with controls to 201±5 mmHg. Albumin excretion increased 235-fold in L-NAME-treated rats. Heart weight index averaged 3.5±0.1 and 3.8±0.1 mg/g body weight (p<0.05) in control and L-NAME rats, respectively, indicating moderate cardiac hypertrophy induced by L-NAME. Preglomerular vascular lesions affected 63±6% of interlobular arteries and 10±2% of afferent arterioles (vs. 8±3 and 0.8±0.4% in controls, respectively). Hydralazine and candesartan cilexetil treatment similarly reduced SBP to 153±7, and 165±6 mmHg, respectively. However, candesartan provided more protection, in terms of no significant change in albuminuria (vs. 25-fold increase with hydralazine), regression of cardiac hypertrophy, frequency of vascular lesions and histological indices of renal injury maintained within control values. In conclusion, candesartan cilexetil prevented L-NAME-exacerbated hypertension and associated cardio-renal injury in young SHR, the beneficial effects exceeding those of hydralazine.

Development of salt-sensitive hypertension in a sensory denervated model: the underlying mechanisms.

We use a novel salt-sensitive hypertensive model recently developed in our laboratory. This model shows that neonatal degeneration of capsaicin-sensitive sensory nerves renders a rat responsive to a salt load with a significant rise in blood pressure (BP). To test the hypothesis that development of salt-sensitive hypertension in sensory denervated rats is mediated by abnormal regulation of both circulating and tissue renin-angiotensin systems (RAS), neonatal Wistar rats were given capsaicin, 50 mg/kg s.c., on the first and second days of life. Control rats were treated with vehicle solution. After the weaning period, male rats were divided into four groups and subjected to the following treatments for three weeks: control + high sodium diet (4%, CON-HS), capsaicin pretreatment + normal sodium diet (0.5%, CAP-NS), capsaicin pretreatment + high sodium diet (CAP-HS), and capsaicin pretreatment + high sodium diet + candesartan cilexetil (10 mg/kg/per day, CAP-HS-CAN). Radioimmunoassay shows that plasma renin activity (ng/ml/hr, PRA) was higher in CAP-NS (2.58±0.17) than in CON-HS (0.14±0.03) and CAP-HS (0.74±0.15), and it was higher in CAP-HS than in CON-HS (p<0.05). Western blot analysis shows that expression of the angiotensin II (Ang II) type 1 (AT1) receptor in both the renal cortex and outer medulla was higher in CAP-HS than in CON-HS and CAP-NS rats (p<0.05). Expression of the Ang II type 2 (AT2) receptor in the renal cortex was higher in both CAP-HS and CAP-NS than in CON-HS rats (p<0.05), but there was no difference in AT2-receptor expression in the renal medulla between CAP-HS, CAP-NS, and CON-HS rats. Likewise, there was no difference in AT1-receptor expression in mesenteric resistance arteries between CAP-HS, CAP-NS, and CON-HS rats. In contrast, mesenteric AT2-receptor expression was lower in CAP-HS than in CAP-NS and CON-HS rats (p<0.05). Tail-cuff systolic BP (mmHg) shows that blockade of the AT1-receptor with candesartan prevents the development of hypertension in CAP-HS rats (by the end of the experiment, CON-HS, 122±3; CAP-NS, 118±10; CAP-HS, 169±9; CAP-HS-CAN, 129±2, p<0.05). Thus, both circulating and tissue RAS in sensory-denervated rats are abnormally regulated in response to a high-salt intake, which may contribute to increased salt sensitivity and account for the effectiveness of candesartan in lowering BP in this model.

Expression of cell cycle proteins in blood vessels of angiotensin II-infused rats: role of AT(1) receptors.

Angiotensin II is an important modulator of cell growth through AT(1) receptors, as demonstrated both in vivo and in vitro. We investigated the role of proteins involved in the cell cycle, including cyclin D1, cyclin-dependent kinase 4 (cdk4), and cyclin-dependent kinase inhibitors p21 and p27 in blood vessels of angiotensin II-infused rats and the effect therein of the AT(1)-receptor antagonist losartan. Male Sprague-Dawley rats were infused for 7 days with angiotensin II (120 ng/kg per minute SC) and/or treated with losartan (10 mg/kg per day orally). DNA synthesis in mesenteric arteries was evaluated by radiolabeled (3)H-thymidine incorporation. The expression of cyclin D1, cdk4, p21, and p27, which play critical roles during the G(1)-phase of the cell cycle process, was examined by Western blot analysis. Tail-cuff systolic blood pressure (mm Hg) was elevated (P<0.01, n=9) in angiotensin II-infused rats (161.3+/-8.2) versus control rats (110.1+/-5.3) and normalized by losartan (104.4+/-3.2). Radiolabeled (3)H-thymidine incorporation (cpm/100 microgram DNA) showed that angiotensin II infusion significantly increased DNA synthesis (152+/-5% versus 102+/-6% of control rats, P<0.05). Expression of cyclin D1 and cdk4 was significantly increased in the angiotensin II group to 213.7+/-8% and 263.6+/-37% of control animals, respectively, whereas expression of p21 and p27 was significantly decreased in the angiotensin II group to 23.2+/-10.4% and 10.3+/-5.3% of control animals, respectively. These effects induced by angiotensin II were normalized in the presence of losartan. Thus, when AT(1) receptors are stimulated in vivo, DNA synthesis is enhanced in blood vessels by activation of cyclin D1 and cdk4. Reduction in cell cycle kinase inhibitors p21 and p27 may contribute to activation of growth induced by in vivo AT(1) receptor stimulation.