RESUMO
PURPOSE: To describe a technique for en bloc harvesting of the corpus cavernosum, cavernous artery and urethra from transplant organ donors and contraction-relaxation experiments with corpus cavernosum smooth muscle. MATERIALS AND METHODS: The corpus cavernosum was dissected to the point of attachment with the crus penis. A 3 cm segment (corpus cavernosum and urethra) was isolated and placed in ice-cold sterile transportation buffer. Under magnification, the cavernous artery was dissected. Thus, 2 cm fragments of cavernous artery and corpus cavernosum were obtained. Strips measuring 3 x 3 x 8 mm(3) were then mounted vertically in an isolated organ bath device. Contractions were measured isometrically with a Narco-Biosystems force displacement transducer (model F-60, Narco-Biosystems, Houston, TX, USA) and recorded on a 4-channel Narco-Biosystems desk model polygraph. RESULTS: Phenylephrine (1 microM) was used to induce tonic contractions in the corpus cavernosum (3-5 g tension) and cavernous artery (0.5-1 g tension) until reaching a plateau. After precontraction, smooth muscle relaxants were used to produce relaxation-response curves (10(-12) M to 10(-4) M). Sodium nitroprusside was used as a relaxation control. CONCLUSION: The harvesting technique and the smooth muscle contraction-relaxation model described in this study were shown to be useful instruments in the search for new drugs for the treatment of human erectile dysfunction.
Assuntos
Endotélio Vascular/fisiologia , Disfunção Erétil/cirurgia , Relaxamento Muscular/fisiologia , Músculo Liso/fisiologia , Pênis/cirurgia , Adulto , Idoso , Disfunção Erétil/fisiopatologia , Humanos , Masculino , Pessoa de Meia-Idade , Modelos Teóricos , Pênis/inervação , Pênis/fisiopatologiaRESUMO
PURPOSE: To describe a technique for en bloc harvesting of the corpus cavernosum, cavernous artery and urethra from transplant organ donors and contraction-relaxation experiments with corpus cavernosum smooth muscle. MATERIALS AND METHODS: The corpus cavernosum was dissected to the point of attachment with the crus penis. A 3 cm segment (corpus cavernosum and urethra) was isolated and placed in ice-cold sterile transportation buffer. Under magnification, the cavernous artery was dissected. Thus, 2 cm fragments of cavernous artery and corpus cavernosum were obtained. Strips measuring 3 x 3 x 8 mm3 were then mounted vertically in an isolated organ bath device. Contractions were measured isometrically with a Narco-Biosystems force displacement transducer (model F-60, Narco-Biosystems, Houston, TX, USA) and recorded on a 4-channel Narco-Biosystems desk model polygraph. RESULTS: Phenylephrine (1µM) was used to induce tonic contractions in the corpus cavernosum (3 - 5 g tension) and cavernous artery (0.5 - 1g tension) until reaching a plateau. After precontraction, smooth muscle relaxants were used to produce relaxation-response curves (10-12M to 10-4 M). Sodium nitroprusside was used as a relaxation control. CONCLUSION: The harvesting technique and the smooth muscle contraction-relaxation model described in this study were shown to be useful instruments in the search for new drugs for the treatment of human erectile dysfunction.
Assuntos
Adulto , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Endotélio Vascular/fisiologia , Disfunção Erétil/cirurgia , Relaxamento Muscular/fisiologia , Músculo Liso/fisiologia , Pênis/cirurgia , Disfunção Erétil/fisiopatologia , Modelos Teóricos , Pênis/inervação , Pênis/fisiopatologiaRESUMO
INTRODUCTION: Endothelial dysfunction characterized by endogenous nitric oxide (NO) deficiency made 56% of patients affected with erectile dysfunction decline treatment with PDE-5 inhibitors. New forms of treatment are currently being developed for this group of patients. MATERIALS AND METHODS: The study compared the effect of sodium nitroprusside (SNP) and two substances of the nitrosyl-ruthenium complex, cis-[Ru(bpy)2(SO3)(NO)]PF-6-9 ("FONO1") and trans-[Ru(NH3)4(caffeine)(NO)]C13 ("LLNO1") on relaxation of rabbit corpus cavernosum smooth muscle and aortic vascular endothelium. The samples were immersed in isolated baths and precontracted with 0.1 microM phenylephrine (PE) and the corresponding relaxation concentration/response curves were plotted. In order to investigate the relaxation mechanisms involved, 100 microM ODQ (a soluble guanylate cyclase-specific inhibitor), 3 microM or 10 microM oxyhemoglobin (an extracellular NO scavenger) or 1 mM L-cysteine (a nitrosyl anion-specific scavenger) was added to the samples. RESULTS: All the NO donors tested produced a significant level of relaxation in the vascular endothelium. In corpus cavernosum samples, FONO1 produced no significant effect, but LLNO1 and SNP induced dose-dependent relaxation with comparable potency (pEC50 = 6.14 +/- 0.08 and 6.4 +/- 0.14, respectively) and maximum effect (Emax = 82% vs. 100%, respectively). All NO donors were found to activate soluble guanylate cyclase, since the addition of the corresponding inhibitor (100 microM ODQ) completely neutralized the relaxation effect observed. The addition of oxyhemoglobin reduced the relaxation effect, but did not inhibit it completely. In aortic vascular endothelium 3 microM oxyhemoglobin decreased the relaxation effect by 26% on the average, while 10 microM oxyhemoglobin reduced it by over 52%. The addition of 100 microM L-cysteine produced no significant inhibiting effect. CONCLUSIONS: These results suggest that LLNO1 and FONO1 are potent vasodilators. LLNO1 was shown to induce a significant level of relaxation in rabbit corpus cavernosum. The substances tested were shown to activate soluble guanylate cyclase and release intracellular NO.
Assuntos
Aorta Torácica/efeitos dos fármacos , Músculo Liso/efeitos dos fármacos , Doadores de Óxido Nítrico/farmacologia , Nitroprussiato/farmacologia , Pênis/efeitos dos fármacos , Compostos de Rutênio/farmacologia , Animais , Endotélio Vascular/efeitos dos fármacos , Masculino , Relaxamento Muscular , CoelhosRESUMO
INTRODUCTION: Endothelial dysfunction characterized by endogenous nitric oxide (NO) deficiency made 56 percent of patients affected with erectile dysfunction decline treatment with PDE-5 inhibitors. New forms of treatment are currently being developed for this group of patients. MATERIALS AND METHODS: The study compared the effect of sodium nitroprusside (SNP) and two substances of the nitrosyl-ruthenium complex, cis-[Ru(bpy)2(SO3)(NO)]PF-6-9 ("FONO1) and trans-[Ru(NH3)4(caffeine)(NO)]C13 ("LLNO1) on relaxation of rabbit corpus cavernosum smooth muscle and aortic vascular endothelium. The samples were immersed in isolated baths and precontracted with 0.1 µM phenylephrine (PE) and the corresponding relaxation concentration/response curves were plotted. In order to investigate the relaxation mechanisms involved, 100 µM ODQ (a soluble guanylate cyclase-specific inhibitor), 3 µM or 10 µM oxyhemoglobin (an extracellular NO scavenger) or 1 mM L-cysteine (a nitrosyl anion-specific scavenger) was added to the samples. RESULTS: All the NO donors tested produced a significant level of relaxation in the vascular endothelium. In corpus cavernosum samples, FONO1 produced no significant effect, but LLNO1 and SNP induced dose-dependent relaxation with comparable potency (pEC50 = 6.14 ± 0.08 and 6.4 ± 0.14, respectively) and maximum effect (Emax = 82 percent vs. 100 percent, respectively). All NO donors were found to activate soluble guanylate cyclase, since the addition of the corresponding inhibitor (100 µM ODQ) completely neutralized the relaxation effect observed. The addition of oxyhemoglobin reduced the relaxation effect, but did not inhibit it completely. In aortic vascular endothelium 3 µM oxyhemoglobin decreased the relaxation effect by 26 percent on the average, while 10 µM oxyhemoglobin reduced it by over 52 percent. The addition of 100 µM L-cysteine produced no significant inhibiting effect. CONCLUSIONS: These results suggest that LLNO1...