Cheng Wen

l-Arginine Prevents Corticotropin-Induced Increases in Blood Pressure in the Rat

In this study we examined whether L-arginine treatment could prevent corticotropin (ACTH)-induced increases in blood pressure in the Sprague-Dawley rat. Sixty rats were randomly divided into six groups (n = 10): sham injection, ACTH injection (0.5 mg/kg per day in divided doses), L-arginine (0.6%) in food plus sham injection, L-arginine plus ACTH treatment, D-arginine (0.6%) in food plus sham injection, and D-arginine plus ACTH. Systolic pressure, water intake, urine volume, body weight, plasma and urinary electrolytes, and serum corticosterone concentrations were measured. ACTH increased systolic pressure (from 127 +/- 2 to 165 +/- 6 mm Hg, P < .001), water intake, and urine volume and decreased body weight body weight. L-Arginine reduced ACTH-induced blood pressure rises (130 +/- 3 mm Hg, P < .001) but had no effect on blood pressure in sham-treated rats. D-Arginine did not affect blood pressure in sham-treated rats, and systolic pressure in D-arginine+ACTH-treated rats was similar to that of ACTH-treated rats. L-Arginine decreased serum corticosterone concentrations in sham-treated rats (424 +/- 42 versus 238 +/- 25 ng/mL, P < .01), but D-arginine had no effect. However, both drugs decreased serum corticosterone concentrations in ACTH-treated rats (1071 +/- 117 versus 739 +/- 95 and 695 +/- 72 ng/mL for L- and D-arginine, respectively; both P < .05). As L-arginine but not D-arginine prevented ACTH-induced increases in blood pressure in Sprague-Dawley rats and both L- and D-arginine reduced serum corticosterone concentrations in ACTH-treated rats, the effects of L-arginine in preventing ACTH-induced hypertension were not simply a consequence of decreased corticosterone secretion.

Decreased Renal Expression of Nitric Oxide Synthase Isoforms in Adrenocorticotropin-Induced and Corticosterone-Induced Hypertension

Abstract—Administration of adrenocorticotropic hormone (ACTH) leads to the development of hypertension. Because glucocorticoids can affect the nitric oxide system at several sites, the present study tested the hypothesis that nitric oxide synthase (NOS) expression may be altered in ACTH-induced and corticosterone-induced hypertension in the rat. This was addressed by measuring Nos1, Nos2, and Nos3 mRNA in the kidney, adrenal gland, heart, and hypothalamus of 16 ACTH-treated and 16 vehicle-treated rats as well as in 10 corticosterone-treated and 10 control rats. In addition, in situ hybridization and immunohistochemistry were used to confirm changes by detection of Nos in RNA and NOS protein in tissues. Systolic blood pressure of ACTH and corticosterone rats was elevated (165±6 and 162±11 mmHg;P <0.001 versus control). Each Nos isoform mRNA was measured by reverse transcriptase-polymerase chain reaction technique. In ACTH rats, mRNA for Nos2 was reduced in renal cortex by 58±5% and in medulla by 68±7%; for Nos3, mRNA reductions of 59±6% and 51±11% were seen (P <0.001 after Hochberg correction for multiple comparisons). In corticosterone rats, Nos2 mRNA decreased in cortex by 68±5% and in medulla by 62±6%;Nos3 mRNA by 50±8% in cortex, and Nos1 by 29±7% in medulla (all P <0.001 after Hochberg correction). Reductions seen in kidney were supported by in situ hybridization and immunohistochemistry. Apart from a 62±2% decrease in Nos2 mRNA in adrenal of ACTH rats (corrected P <0.05), no significant changes were seen in the other nonrenal tissues for any isoform. In conclusion, we have shown for the first time that the physiological components of glucocorticoid action (ACTH and corticosterone) when given chronically in vivo reduce Nos2 and Nos3 expression in the kidney. Such changes are consistent with a role in hypertension for ACTH and corticosterone.

Validation of transonic small animal flowmeter for measurement of cardiac output and regional blood flow in the rat.

The objective of the present study was to validate a transonic flowmeter system with two probes (model 3SS for cardiac output (CO) and 1RB for organ flows) in Sprague-Dawley (SD) rats first by measuring blood flow through pump-infused isolated renal artery and ascending aorta, and then through measurements of CO and renal, mesenteric, and hindquarter blood flow (RBF, MBF, HBF) in vivo. We measured in vivo baseline flow and changes in flow induced by dopamine and propranolol for CO, prostaglandin E2 (PGE2), and angiotensin II (AII) for RBF and pentobarbital sodium for MBF and HBF. Correlations between meter and pump flow were linear (r = 0.999, p < 0.001) with close agreement both in ascending aorta and renal artery flow measurements. The baseline values were 15 +/- 0.7 ml/100 g/min for CO, 4 +/- 0.1 ml/100 g/min for RBF, 7 +/- 0.3 ml/100 g/min for MBF, and 6 +/- 0.3 ml/100 g/min for HBF, respectively. The system reliably detected increase and/or decrease in CO and regional blood flows. The transonic flowmeter system is accurate, highly reproducible, and compatible with other established techniques for measuring CO and regional blood flows in the rat.

Adrenocorticotrophin-induced hypertension: effects of mineralocorticoid and glucocorticoid receptor antagonism.

OBJECTIVE To examine whether the increase of blood pressure in adrenocorticotrophin-treated rats is mediated through mineralocorticoid or glucocorticoid receptors or corticosterone 6 beta-hydroxylation inhibition. DESIGN Rats were randomly allocated to 14 treatment groups for 10 days. The treatments included sham injection (n = 35), adrenocorticotrophin (5, 100, 500 micrograms/kg per day, subcutaneously, n = 5, 15 and 15, respectively), spironolactone (100 mg/kg per day, subcutaneously, n = 15), standard-dose or high-dose RU 486 (70 mg/kg every 3 days or 70 mg/kg per day, subcutaneously, n = 5 and 10, respectively), spironolactone + adrenocorticotrophin (100 micrograms/kg per day, n = 5, or 500 micrograms/kg per day, n = 10), standard-dose RU 486 + adrenocorticotrophin (500 micrograms/kg per day, n = 5), high-dose RU 486 + adrenocorticotrophin (100 micrograms/kg per day, n = 10), troleandomycin (40 mg/kg per day, subcutaneously, n = 5) and troleandomycin + adrenocorticotrophin (5 micrograms/kg per day, n = 5). Systolic blood pressure and metabolic parameters were measured every second day. RESULTS Adrenocorticotrophin treatment increased systolic blood pressure dose-dependently (5 micrograms/kg per day: +14 +/- 2 mmHg; 100 micrograms/kg per day: +20 +/- 2 mmHg; 500 micrograms/kg per day: +28 +/- 2 mmHg, all P < 0.001). Adrenocorticotrophin at 100 and 500 micrograms/kg per day increased plasma sodium and decreased plasma potassium concentrations. Spironolactone did not block adrenocorticotrophin-induced systolic blood pressure changes but did block changes in plasma sodium and potassium levels. Standard-dose RU 486 did not modify the adrenocorticotrophin-induced (500 micrograms/kg per day) systolic blood pressure rise but blocked the effect of adrenocorticotrophin on body weight. High-dose RU 486 partially blocked the adrenocorticotrophin-induced (100 micrograms/kg per day) systolic blood pressure increase (adrenocorticotrophin at 100 micrograms/kg per day: 143 +/- 3 mmHg; high-dose RU 486 + adrenocorticotrophin at 100 micrograms/kg per day: 128 +/- 5 mmHg, P < 0.001) and body-weight loss. Troleandomycin did not alter the development of adrenocorticotrophin-induced hypertension. CONCLUSIONS Spironolactone and standard-dose RU 486 did not modify adrenocorticotrophin-induced hypertension despite demonstrable antimineralocorticoid and antiglucocorticoid actions. High-dose RU 486 partially blocked adrenocorticotrophin-induced (100 micrograms/kg per day) hypertension, suggesting either a permissive effect of glucocorticoid on blood pressure or other antihypertensive actions of RU 486. Inhibition of glucocorticoid 6 beta-hydroxylation by troleandomycin did not modify adrenocorticotrophin-induced hypertension, suggesting that effects of corticosterone 6 beta-hydroxylation in adrenocorticotrophin-induced hypertension are negligible.

Adrenocorticotrophin dose-response relationships in the rat: haemodynamic, metabolic and hormonal effects.

Objective To determine adrenocorticotrophin dose–response relationships for increase of blood pressure and metabolic parameters of the Sprague–Dawley rat. Methods We injected 120 male Sprague–Dawley rats twice daily subcutaneously for 10 days with 0.5, 1, 5, 50, 100, 200 or 500 μg/kg synthetic adrenocorticotrophin per day (all n = 10) or subjected them to sham injection (0.9% NaCl; n = 50). Systolic blood pressure, 24 h food intake, water intake, urine volume and body weight were measured. Data from a further 45 rats treated with 500 μg/kg per day adrenocorticotrophin in previous studies were included in the blood pressure analyses. After we had killed these rats, their organ weights (kidney, heart, adrenal) and plasma electrolyte, adrenocorticotrophin and serum corticosterone concentrations were measured. Results On the final day of treatment systolic blood pressure of sham-injection control rats was 123 ± 1 mmHg (n = 50). Compared with sham treatment, a low dose of adrenocorticotrophin (1 μg/kg per day) increased systolic blood pressure from 122 ± 1 to 130 ± 2 mmHg (P < 0.001) without any metabolic effects, whereas a high dose of adrenocorticotrophin (500 μg/kg per day) increased systolic blood pressure from 121 ± 1 to 150 ± 2 mmHg (P < 0.001, n = 55) with increases in intake of water and urine volume (P < 0.001, n = 10) and a decrease in body weight (P < 0.001, n = 10). Plasma adrenocorticotrophin and serum corticosterone concentrations for the sham-injection control group were 162 ± 12 pg/ml (36 ± 3 pmol/l) and 376 ± 18 ng/ml (1038 ± 50 nmol/l), respectively. Plasma adrenocorticotrophin concentration was elevated by injections of 100 (P < 0.05), 200 (P < 0.01) and 500 μg/kg adrenocorticotrophin per day (P = 0.001). Serum corticosterone concentration was not significantly different from that of sham-injection rats with 0.5–5 μg/kg adrenocorticotrophin per day but was increased by injection of 50–500 μg/kg adrenocorticotrophin per day (P < 0.001). Conclusions These results define 1 μg/kg adrenocorticotrophin per day, administered subcutaneously, as the threshold dose for causing a rise in blood pressure in the rat. Thus administration of adrenocorticotrophin increases systolic blood pressure at doses that induce minimal adrenocorticotrophin metabolic effects. Administration of a low dose of adrenocorticotrophin to the rat is a suitable model for stress-induced hypertension. J Hypertens 16:593–600

Haemodynamic mechanisms of corticotropin (ACTH)-induced hypertension in the rat.

OBJECTIVES To investigate the roles of cardiac output and systemic and regional resistances in corticotropin (ACTH)-induced hypertension in the rat METHODS This study consisted of three series of experiments with eight groups of male Sprague-Dawley rats (n = 132). Series 1 comprised groups 1-4, where group 1 = sham (0.9% NaCl, subcutaneous (s.c.) injection); group 2 = ACTH (0.5 mg/kg per day, s.c.); group 3 = atenolol + sham; group 4 = atenolol + ACTH treatments. Series 2 comprised groups 5 and 6, where group 5 = minoxidil + sham and group 6 = minoxidil + ACTH treatments. Series 3 comprised groups 7 and 8, where group 7 = ramipril + sham and group 8 = ramipril + ACTH treatments. Systolic blood pressure, water and food intakes, urine volume, and body weight were measured every second day. After 10 days of treatment, mean arterial blood pressure was measured by intra-arterial cannulation, and cardiac output (CO), and renal, mesenteric and hindquarter blood flows (RBF, MBF and HBF) determined using transonic small animal flowmeters. RESULTS ACTH treatment increased blood pressure (P < 0.001) with a rise in CO (P < 0.01) and renal vascular resistance (RVR, P < 0.05), but did not affect total peripheral resistance (TPR). Atenolol blocked the rise in CO without affecting the rise in blood pressure produced by ACTH treatment Minoxidil lowered TPR, but did not prevent the rise in blood pressure or renal vascular resistance. Ramipril blunted the rise in RVR and blood pressure without significantly affecting TPR. CONCLUSION Neither preventing rise in CO nor lowering TPR altered the ACTH-induced rise in blood pressure in the rat However, both the hypertension and rise in RVR were prevented by ramipril. These data suggest that increase in RVR may play a role in the pathogenesis of ACTH-induced hypertension in the rat.

Hemodynamic profile of corticotropin-induced hypertension in the rat

Objectives To examine hemodynamic variables in corticotropin-induced hypertension in rats and the effects of reversal of the hypertension by L-arginine on the hemodynamic profile. Methods Sixty male Sprague–Dawley rats were randomly divided into four groups: sham treatment (0.9% NaCl, injected subcutaneously); 0.5 mg/kg corticotropin per day, subcutaneously; 0.6% L-arginine in food plus sham; and L-arginine plus corticotropin. Systolic blood pressure, water and food intakes, urine volume, and body weight were measured every second day. After 10 days mean arterial blood pressure was measured by intra-arterial cannulation, and cardiac output, and renal, mesenteric, and hindquarter blood flows were determined using transonic small animal flowmeters. Results Injection of corticotropin increased blood pressure, water intake, urine volume, and plasma sodium concentration, and decreased body weight and plasma potassium concentration. It increased cardiac output (P < 0.01), mesenteric blood flow (P < 0.05), and renal vascular resistance (P < 0.05), and decreased renal blood flow (P < 0.05), but did not change calculated total peripheral resistance, hindquarter blood flow, mesenteric or hindquarter vascular resistance. L-arginine prevented corticotropin-induced rises in blood pressure (P < 0.001) and renal vascular resistance (P < 0.05), and a fall in renal blood flow (P < 0.05), but did not affect other hemodynamic variables. Conclusion The hemodynamic profile of corticotropin-induced hypertension in the rat is characterized by a rise in cardiac output and renal vascular resistance, a fall in renal blood flow, but no change in total peripheral resistance, hindquarter blood flow, mesenteric vascular resistance, or hindquarter vascular resistance. L-arginine prevented corticotropin-induced rises both in blood pressure and in renal vascular resistance in the rat. These data suggest that the increase in renal vascular resistance might play a role in corticotropin-induced hypertension in the rat.

LONG-TERM OUABAIN ADMINISTRATION DOES NOT ALTER BLOOD PRESSURE IN CONSCIOUS SPRAGUE-DAWLEY RATS

1. We tested the ability of ouabain to cause chronic hyper tension by continuously infusing ouabain for 28 days (mini‐osmotic pump implantation; i.p.). The blood pressure and metabolic effects of sham (150 mmol/L NaCI; n= 12) or ouabain infusion (10 μg/kg per day; n= 14; 100 μg/kg per day; n = 14) were examined in conscious Sprague‐Dawley rats.

DEHYDROEPIANDROSTERONE DOES NOT PREVENT ADRENOCORTICOTROPHIN‐INDUCED HYPERTENSION IN CONSCIOUS RATS

1. We tested the hypothesis that dehydroepiandrosterone (DHEA), which prevents dexamethasone‐induced hypertension in rats, may block adrenocorticotrophin (ACTH) hypertension, which has been presumed due to corticosterone excess. The blood pressure and metabolic effects of DHEA (18 mg/kg per day) were examined in sham and ACTH‐treated (0.5 mg/kg per day) conscious Sprague‐Dawley rats (n= 20).

Systemic and regional hemodynamics in cyclosporin A hypertension in the rat

SUMMARY: This study examined the hemodynamic effects of 21 days oral administration of cyclosporin A (CyA) in the male Wistar rat. Forty rats were randomly divided into four groups (n=10). Group 1: Sham (olive oil 1 mL/kg/day by gavage) with cardiac output (CO) measurement. Group 2: CyA (CyA 15 mg/kg/day in olive oil by gavage) with CO measurement. Group 3: Sham as for group 1 with regional flow measurements. Group 4: CyA as for group 2 with regional flow measurements. Systolic blood pressure was measured every fourth day. After 21 days mean arterial blood pressure was measured by intra‐arterial cannulation, and CO, and renal, mesenteric, and hindquarter blood flows (RBF, MBF, and HBF) were determined using transonic small animal flowmeters. Total peripheral resistance (TPR) and regional resistances were calculated. Oral CyA produced a sustained rise in systolic blood pressure. Olive oil did not affect blood pressure. CyA increased TPR (P<0.05), renal vascular resistance (RVR, P<0.01) and MBF (P<0.01), decreased RBF (P<0.01), but did not change CO, HBF, mesenteric or hindquarter vascular resistance. We conclude that chronic oral administration of CyA produces hypertension with a hemodynamic profile characterized by rises in TPR and RVR without changes in CO or mesenteric or hindquarter vascular resistances in the Wistar rat.