Alexandra Oudot

Combination of BAY 60-4552 and vardenafil exerts proerectile facilitator effects in rats with cavernous nerve injury: a proof of concept study for the treatment of phosphodiesterase type 5 inhibitor failure.

BACKGROUND Radical prostatectomy (RP) is frequently responsible for erectile dysfunction (ED). Post-RP patients often show a failure to respond to phosphodiesterase type 5 (PDE5) inhibitors. OBJECTIVE The acute effect of BAY 60-4552, the soluble guanylate cyclase (sGC) stimulator, and vardenafil were evaluated alone or in combination on erectile responses to electrical stimulation of the cavernous nerve (ES CN) in rats with cavernous nerve (CN) crush injury-induced ED. DESIGN, SETTING, AND PARTICIPANTS Male adult Sprague-Dawley rats underwent laparotomy (sham, n=10) or bilateral CN crush injury (n=56). After 3 wk of recovery, erectile function was evaluated under urethane anaesthesia following ES CN at different frequencies. MEASUREMENTS The acute effects of intravenous (IV) injection of vehicle, vardenafil 0.03 mg/kg, BAY 60-4552 0.03 mg/kg or 0.3 mg/kg, or a BAY 60-4552 0.03 mg/kg plus vardenafil 0.03 mg/kg combination were evaluated in CN-crushed rats. RESULTS AND LIMITATIONS Bilateral CN crush injury followed by a 3-wk recovery period decreased erectile responses to ES CN by about 50%. In CN-crushed rats, IV vardenafil 0.03 mg/kg and BAY 60-4552 (0.03 or 0.3 mg/kg) increased erectile responses to ES CN to the same extent: Δ intracavernosal pressure/mean arterial pressure (ICP/MAP) at 10 Hz ES CN was 21±1% after vehicle, 25±3% (p<0.001) after vardenafil, and 26±5% and 27±5% after BAY 60-4552 0.03 mg/kg (p<0.01) and 0.3 mg/kg (p<0.001), respectively. The combination of vardenafil with BAY 60-4552 in CN-crushed rats totally restored erectile responses to ES CN equivalent to sham rats (ΔICP/MAP at 10 Hz ES CN: 34±4% after BAY 60-4552/vardenafil combination vs 39±4% in sham rats; not significant). CONCLUSIONS The present study supports the concept that the combined administration of a sGC stimulator, BAY 60-4552, and vardenafil provides synergistic beneficial effects and might therefore salvage patients who experience treatment failures with PDE5 inhibitors after RP.

ORIGINAL RESEARCH—BASIC SCIENCE: Erectile Dysfunction in Hypercholesterolemic Atherosclerotic Apolipoprotein E Knockout Mice

INTRODUCTION Erectile dysfunction (ED) and cardiovascular diseases share the same risk factors. Although the use of hypercholesterolemic rabbit models has proven to be useful to illustrate the link between ED and hypercholesterolemia, the cost of daily maintenance of the animals and necessity for important amounts of drug have limited their use. AIM We aimed to develop a new model of atherosclerosis-associated ED in a well-known experimental model of atherosclerosis, the apolipoprotein E knockout (ApoE KO) mouse. METHODS Erectile function was evaluated by recording frequency-dependent increases in intracavernous pressure following electrical stimulation of the cavernous nerve in 26-, 32-, and 38-week-old ApoE KO mice fed a Western-type diet and age-matched C57BL6/J anesthetized mice. Atherosclerotic lesions were evaluated by planimetry in oil red O-stained aortas. RESULTS We found that in contrast to C57BL6/J mice, ApoE mice displayed atherosclerotic lesions covering 22% of the aortic luminal surface at 26 weeks of age and increasing to 27% and 35% at 32 weeks and 38 weeks of age, respectively. The amplitude of erectile responses to electrical stimulation of the cavernous nerve was markedly impaired in 26-week-old ApoE KO mice as compared with age-matched C57BL6/J mice. Impairment in erectile function persisted in ApoE KO mice 32 and 38 weeks of age. CONCLUSIONS The ApoE KO mouse, a well-characterized model to study disorders associated with hypercholesterolemia and atherosclerosis in cardiovascular research, could therefore be suitable for investigation of disease-modifying effects of new therapeutic strategies aiming to target both atherosclerosis and ED.

Impact of a Long-Term Sildenafil Treatment on Pressor Response in Conscious Rats With Insulin Resistance and Hypertriglyceridemia

BACKGROUND Insulin resistance constitutes a risk factor for endothelial dysfunction and subsequent cardiovascular diseases including hypertension. Daily treatment with phosphodiesterase type 5 (PDE5) inhibitors has beneficial effects on endothelial function in men with increased cardiovascular risk. Endothelium-dependent vasomotor function is ultimately linked to blood pressure (BP) regulation. We postulated that sildenafil would ameliorate BP and biological markers of endothelial function in fructose-fed rats (FFRs). METHODS Wistar rats were fed a standard chow or a 60% fructose-enriched diet containing 12% fat for 8 weeks (FFR). From week 6 through 8, sildenafil (twice a day subcutaneously, 20 mg/kg) was administered followed by a 1-week washout period. At the end of the washout period, BP was recorded using radiotelemetry following cumulative infusion of norepinephrine (from 50 to 400 ng/kg/min). RESULTS FFR displayed both an impaired glucose tolerance and elevated triglyceridemia. The latter was corrected by sildenafil treatment. Resting BP was similar in all rats, whereas pressor responses were significantly enhanced in FFR (maximal increase in mean BP to norepinephrine: 25.6 +/- 3.8 vs. 40.8 +/- 4.0 mm Hg, P < 0.05) and normalized by sildenafil treatment (24.9 +/- 5.3 mm Hg, not significant vs. control). Urinary levels of 8-isoprostanes and thromboxane B(2) were increased in FFR and corrected by sildenafil treatment. CONCLUSION Thus, chronic treatment with sildenafil normalized BP regulation in an experimental model of insulin resistance and hypertriglyceridemia while restoring normal excretion of urinary biological markers of oxidative stress and cyclooxygenase-derived vasoconstrictors. The modulation of ROS and cyclooxygenase-derived vasoconstrictors generation by a chronic treatment with sildenafil may represent an added benefit beyond PDE5 inhibition.

A new experimental rat model of erectile dysfunction and lower urinary tract symptoms associated with benign prostatic hyperplasia: the testosterone-supplemented spontaneously hypertensive rat

Whats known on the subject? and What does the study add?

How Does Chronic Sildenafil Prevent Vascular Oxidative Stress in Insulin-Resistant Rats?

INTRODUCTION Insulin resistance features both endothelial dysfunction and increased oxidative stress. Both disorders are targeted by a chronic treatment with sildenafil. However, the mechanism of action by which chronic sildenafil exerts its effects on reactive oxygen species sources is still largely unknown. AIM We therefore investigated how chronic sildenafil administration could impact vascular endothelial NO and superoxide release in a rat model of insulin resistance induced by fructose overload. METHODS Adult male Wistar rats were fed a fructose-enriched diet (fructose-fed rats [FFR]) for 9 weeks. From weeks 6-8, sildenafil was administered subcutaneously twice daily (20 mg/kg), followed by a 1-week washout. MAIN OUTCOME MEASURES Vascular endothelial NO and superoxide release were monitored in vitro in thoracic aortic segments using oxidative fluorescence. Specific inhibitors were used to distinguish the respective role of the main superoxide-producing systems within the vascular wall (i.e., mitochondrial respiratory chain and NADPH oxidases). The levels of expression of eNOS, Akt, and NADPH oxidase subunits were determined in the abdominal aorta. RESULTS Chronic sildenafil administration corrected hyperglycemia, hyperinsulinemia, and hypertriglyceridemia in FFR. Moreover, after 9 weeks of diet, while global unstimulated aortic endothelial NO and superoxide release were unchanged in FFR, the relative contribution of the mitochondrial respiratory chain and NADPH oxidases was modified. Chronic sildenafil treatment, even after the 1-week washout period, was able to increase endothelial NO release independently of Akt-dependent phosphorylation by up-regulating eNOS expression, and restored the relative contribution of each superoxide-producing system examined, yielding endothelial superoxide release. Finally, in vitro incubation of aortic segments with sildenafil markedly decreased the endothelial aortic superoxide release. CONCLUSIONS The present study showed that chronic sildenafil produced sustained vascular antioxidant effects in insulin-resistant rats by increasing NO release and regulating vascular superoxide release, supporting therefore further investigations using chronic sildenafil administration in preventing cardiovascular alterations associated with oxidative stress.

Experimental evidence that a combination of sGC stimulator BAY 60-4552 and PDE5 inhibitor vardenafil could salvage ED-patients with insufficient response to PDE5 inhibitors after cavernous nerve injury

Background Radical prostatectomy (RP) is frequently responsible for erectile dysfunction (ED). Post-RP patients often show insufficient response or treatment failures to PDE5 inhibitor therapy. This study was undertaken to evaluate the acute effects of the soluble guanylate cyclase (sGC) stimulator, BAY 60-4552 and vardenafil administered alone or in combination on erectile responses to electrical stimulation of the cavernous nerve (ES-CN) in rats with cavernous nerve (CN) crush injury-induced ED.

Chronic inhibition of cyclic GMP phosphodiesterase 5A prevents and reverses endothelial dysfunction and oxidative stress associated with the metabolic syndrome

Materials and methods Wistar rats (n = 10–14 per group) were fed a standard chow (CONT) or a 60% fructose/5% fat (% by weight)enriched diet for 8 weeks (FFR). From week 5 through 8, sildenafil was administered twice a day (sc, 20 mg/kg, FFR+SIL), thus reaching clinically relevant plasma concentrations circa 20 nM unbound known to give efficacy in man (Pfizer Inc., data on file), then a 1-week wash-out period from sildenafil was observed. Isometric tension studies were performed on isolated aortic and superior mesenteric arterial (SMA) rings precontracted with noradrenaline to build concentration-response curves (CRC) to endothelium-dependent (ACh and A23187) and -independent (SNP) relaxants in presence of indomethacin. Urinary 8-isoprostanes (IPT) and plasma levels of IL-6 and TNF-α were also evaluated. Results Relaxations to ACh were reduced in aortas of FFR (10-5 M: -102.6 ± -2.4% vs -89.2 ± 4.7, p < 0.001) while only slightly affected in SMA rings. Relaxations to A23187 were significantly reduced both in aortic and SMA rings of FFR. In aortas, sildenafil treatment restored normal endothelium-dependent relaxations to ACh (10-5 M: -104.2 ± 3.0%, p < 0.001) even after one week of wash-out from treatment. In SMA rings, a leftward shift of the CRC to ACh could be detected (pD2: -8.12 ± 0.11 vs -8.60 ± 0.08, p < 0.05). Relaxations to A23187 were also restored by sildenafil in both aortic and SMA rings of FFR. Enhanced compensatory endothelium-independent relaxations to SNP in FFR were not modified by sildenafil treatment. Neither IL-6 nor TNF-α were modified by the fructose or sildenafil treatment. Urinary IPT levels was normalized by the sildenafil treatment (FFR: 2.07 ± 0.36 vs CONT:0.95 ± 0.14 vs FFR+SIL: 0.88 ± 0.13 ng/ml/24 h, p < 0.05).

Daily treatment with sildenafil, an inhibitor of cGMP-dependent phosphodiesterase, normalizes the exaggerated hemodynamic response to norepinephrine in fructose-fed rats

Background Enhanced responsiveness of the vascular smooth muscle cell to vasoconstrictor stimuli has been described in insulin-resistant states. Fructose overload leads to the development of the metabolic syndrome and its associated cardiovascular disorders including hypertension. We postulated that chronic treatment with sildenafil would regulate the abnormally increased pressor response to a norepinephrine challenge in fructose-fed rats. Material and methods Wistar rats were fed a standard chow (CONT, n = 12) or a 60% fructose-enriched diet containing 5% fat (% by weight) for 8 weeks (FFR, n = 12). From week 5 through 8, sildenafil (twice a day sc, 20 mg/kg) was administered (FFR+SIL, n = 14), thus reaching clinically relevant plasma concentrations circa 20 nM unbound (Pfizer Inc., data on file), followed by a 1-week wash-out period (W9). On W9, in a sub-group of these conscious rats (n = 5) that had previously been implanted with telemetric devices, AP was recorded following cumulative infusion of noradrenaline (NA 50, 100, 200, 400 ng/kg/min). In all rats, urinary 8-isoprostane (IPT) and TxB2 levels were determined during a 24-hour period. Tissue ET-1 content was measured in segments of aortic and mesenteric artery. Results Fasting glycemia was unchanged in all rats while triglyceridemia were elevated in FFR compared to CONT (2.0 ± 0.4 mM vs 1.2 ± 0.2, p < 0.05) and corrected by sildenafil treatment (1.3 ± 0.2, NS vs CONT). Resting AP was similar in all rats. Pressor responses to NA were exacerbated in FFR (maximal increase from basal MAP: +22.8 ± 1.7 vs +38.0 ± 7.3 mmHg, respectively, p < 0.05) and normalized by sildenafil treatment (+24.9 ± 5.2 mmHg, NS vs CONT). Vascular ET-1 levels were not modified by the fructose nor sildenafil treatment whereas the increased production of urinary IPT and TxB2 in FFR versus CONT (2.07 ± 0.36 ng/ml/24 h vs 0.88 ± 0.13 and 12.86 ± 1.91 pg/ml/24 h vs 7.25 ± 0.59, respectively, p < 0.05) were corrected following sildenafil treatment (0.95 ± 0.14 ng/ ml/24 h and 8.74 ± 1.06 pg/ml/24 h, respectively, NS vs CONT).