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237 Relaxant effect of ferulago syriaca root extract on human corpus cavernosum
237 RELAXANT EFFECT OF FERULAGO HUMAN CORPUS CAVERNOSUM Oztiirk B.‘, Giir S.>, Coskun Cetinkaya M.’
SYRIACA
M.3, Kosan M.4, Erdurak
ROOT
EXTRACT
ON
C5, Hafez G.6, Ozgunes
238 A S-HYDROXYTRYPTAMINE PONSE IN RABBIT CAVERNOSAL TO THE ERECTILE PROCESS
BIPHASIC RESMUSCLE: RELEVANCE
0.6, m’,
‘Ankara Numune Education and Research Hospital, 2nd Urology Clinic, Ankara, Turkey, *Faculty of Pharmacy, Ankara University, Department of Pharmacology, Ankara, Turkey, 3Faculty of Pharmacy, Ankara University, Department of Pharmaceutical Botany, Ankara, Turkey, 4Ankara Numune Education and Research Hospital, 32nd Urology Clinic, Ankara, Turkey, 5Faculty of Pharmacy, Ankara University, 2department of Pharmaceutical Botany, Ankara, Turkey, 6Faculty of Pharmacy, Ankara University, ldepartment of Pharmacology, Ankara, Turkey INTRODUCTION & OBJECTIVES: The effects of Fen&go which has been used for its aphrodisiacs properties in Turkey human corpus cavemosum.
(5-HT)-MEDIATED SMOOTH
s)~iaca root extract were investigated on
MATERIAL & METHODS: Strips of human corpus cavernosum were precontracted with phenylephrine (10 mM). Relaxations induced by extract between 1.8- 9.0 mg doses were studied in strips of corpus cavernosum in which tone was elicited by phenylephrine (10 mmoV1). After washing period tissues were incubated with NOsynthase inhibitor, L-NAME or selective soluble guanylate cyclase inhibitor, lH-1.2.4oxadiazolo [4,3-alquinoxalin-lone (ODQ) for 30 min. Furthermore, effects of extract on EFS, acetylcholine, forskolin and SNP- induced relaxation were also evaluated in human corpus cavemosum strips. RESULTS: Extract caused long-lived and dose-dependent relaxations (17.2 + 0.9%, 41.4 i 3.8 %, 60.5 i 3.2%, 86.7+ 2.8% at doses of 1.8, 3.6, 5.4 and 9 mg respectively) in precontracted corporal tissues. The nitric oxide synthesis inhibitor NG-nitro-Larginine methyl ester (L-NAME; 10 mM) produced significant inhibition at the low doses (12.7 + 1.5%, 25.5 & 7.1 at doses of 1.8 and 3.6) although there was no suppression at high doses of extract in the organ bath. The soluble guanylate cyclase inhibitor ODQ (30 mM) significantly affected the extract-induced relaxation (at 9 mg dose of extract, 86.7 +2.8% before, and 35.6 54.95% after ODQ treatment, p
Thompson
C2, Mikhailidis
D.2, Morgan R.3
‘Royal Free Hospital and University College Medical School, Royal Free Campus, Department of Urology and Clinical Biochemistry, Mitcham, United Kingdom, 2Royal Free Hospital and University College Medical School, Royal Free Campus, Department of Clinical Biochemistry, Mitcham, United Kingdom, 3Royal Free Hospital and University College
Medical School, Royal Free Campus, Department of Urology,
Mitcham, United Kingdom
INTRODUCTION & OBJECTIVES: It is of interest that Ketanserin, a S-HT, antagonist and Doxazosin. an aloha-l-blocker which has been shown to have a 5-HT inhihitorv action, were shbwn to ‘have a beneficial effect in the treatment of erectile dysfunction (ED)
(human studies). 5.HT was shown to cause contractions in penile arteries and veins of rat and human, respectively. It is likely that 5-HT might have a similar effect in corpus cavemosal smooth muscle. Therefore, 5-HT might have a role to play on the erectile process as well as the pathogen&s of ED as circulating 5-HT levels are raised in patients with diabetes or peripheral
vascular disease-two
conditions
associated with an increased risk of ED.
MATERIAL & METHODS: The effect of 5-HT on rabbit cavemosal tissues were assessed in organ bath studies. Ketanserin, Doxazosin, Ondansetron and Y-12530 (both 5-HT3 antagonists) were also used to further characterise the receptor profile. Results are presented as median and range (mg/mg=contractionlmg of tissue).
Consistent 5-HT-mediated (10m3M) contractions from baseline recordings were in rabbit cavernosal tissues (n=125). The median contraction was 68.8 mgimg.
RESULTS:
demonstrated
Ketanserin (I Oe5M) completely abolished the contractile response. These contractions were also inhibited by Doxazosin ( 10m4and 10-6M, n=8) in a concentration -dependent manner (1 O4M; 87% reduction, 10-6M; 63.3% reduction). The Doxazosin response was not attributable to
alpha blockade, since alpha-l and 2 antagonists (corynanthine
and yohimbine)
had no effect
on the 5-HT-induced contractions. A biphasic response of transient relaxation followed by substained contraction was observed in 30% of the 5-HT-mediated responses (38 strips of a total of 125). The transient relaxation (median = -20.9 mg/mg; the minus sign = relaxation) was completely abolished by Ondansetron and Y-12530. CONCLUSIONS: 5-HT may have a role to play on the erectile process via its 5-HT,mediated contractile and 5-HT.-mediated relaxant activities. The overall 5.HT induced contractions may represent an in&play between the two S-HT-mediated opposing forces. It is possible that the faster onset 5.HT,-mediated relaxant force modulates the contractile response mediated by activation of the 5-HT,,receptor subtype. The fact that the transient
relaxation is not always present suggests a varlatlon in the rabbit population relating to the erectile 5-HT-mediated response. Our findings further imply that Ketanserin and Doxazosin may be beneficial in the treatment of ED as part of a multi-therapy regime (when a monotherapy failed) via 5-HT dependent mechanisms (rather than exclusively via alpha-l-
adrenergic blockade in the case with Doxazosin).
239 IN VITRO EFFECT OF DIFFERENT DRUGS ON THE VIABILITY ENDOTHELIAL CELLS WA.‘,
INTRACAVERNOUS OF HUMAN
VASOACTIVE CAVERNOSAL
Schultheiss D.‘, Gabouev A.l, Schlote N.l, Wefer J.‘: Mertsching H.Z, Sohn M.3, Stief
240 POLYMORPHISM GENE IN PATIENTS
OF ENDOTHELIAL WITH ERECTILE
Erkan E.‘, Muslumanoglu Kadioglu A.3
A.Y.‘,
Merder
NITRIC OXIDE DYSFUNCTION E.‘, Tefekli
A.‘,
Akbas
SYNTHASE F.2, Ozbek
U.*,
C.G.‘, Jonas U.’
‘Hannover Medical School, Department ofurology, Biotechnology and Artificial Organs, Lebao, Department of Urology, Frankfurt, Germany
Haonover, Germany, 2Leibniz Laboratories for Hannover, Germany, ?St. Markus Hospital,
INTRODUCTION & OBJECTIVES: The endothelium of the corpus cavemosum plays an important role in signal transduction of penile erection. It is directly exposed to intracavemously injected vasoactive agents, which are then passed through the endothelial layer reaching the stromal smooth muscle cells. We investigated commercially available prostaglandin El (Cave+&‘), standard bi-mix of papaverineiphentolamine (AndroskatO) and tri-mix of these agents in physiological dilutions using cultured human cavemosal endothelial cells and fibroblasts. MATERIAL & METHODS: Primary corpus cavemosum endothelial and tibroblast cell cultures were obtained from potent patients (n=12) undergoing penile surgery. 5~10~ cells/well seeded in sixwell plates were exposed for 30min to physiological dilutions of 2Ob.g PGEl (=group P). 30mg papaverineilmg phentolamine (=group A) or t&mix of these agents (=group T) each dissolved in 5m1, lOm1, 2Oml or 5Oml NaCl (= P5 and P50 for the PGEl group etc.). Lactate dehydrogenase (LDH) release as a cytotoxicity marker was measured 6h after drug exposure. The metabolic cell activity was quantified after 48h with the tetrazolium salt-based (MTS) assay and viable cells were counted by performing a Liveil)eada staining. RESULTS: The release of LDH over the first 6h was elevated up to 5-fold (p
‘Haseki Teaching and Research Hospital, Urology, Istanbul, Turkey, University, Medical Faculty, The Institute of Research for Experimental, Turkey, Wanbul University, Medical Faculty, Urology, Istanbul, Turkey
INTRODUCTION & OBJECTIVES: Erectile dysfunction (ED) is a phenomenon which is strictly related to demographic and environmental factors, but the influence of genetic determinants are still not clear. In this trial, we tried to assess the possible role of the polymorphism of endothelial nitric oxide synthase (eNOS) gene on the development and severity of ED. MATERIAL & METHODS: Thirty patients, with a mean age of 56.457.8 (range: 3974) years, who had registered to our andrology policlinic with ED between Febmary 2003 and June 2003 were included in this trial. As the control group, 25 voluntary males, with a mean age of 58.7*9.9 (range: 47-72) years, were included. After a comprehensive physical examination and biochemical evaluation for the risk factors of ED, all cases were searched for the presence of polymorphism of eNOS gene. Detection of the polymorphism was performed in the 4th intron of 7th somatic chromosome which is encoding the eNOS gene, by polymerase chain reaction (PCR). The men with ED were assessed with pharmacopenile dupplex ultrasonography (PPDU) while the potency of the control group was established on the basis of International Index of Erectile Function (IIEF) query. The results of the two groups were compared. For statistical evaluation; chi-square and Student-t tests were used. RESULTS: mellitus and 0.001; 0.023 insufficiency insufficiency eNOS4aib
This effect was less pronounced in the tibroblast cultures where the decrease of viable cells in the more concentrated A and T arouos was less intense after 48h than in endothelial cells (o. The above described results w&e comparably seen in the cell metabolic activity ass& Esp.&ially papaverlneiphentolamine (A) and hi-mix (T) likewise provoked a decrease in activity. CONCLUSIONS: These in vitro data clearly demonstrate the decrease of endothelial cell viabilitv under the influence of intracavemous vasoactive agents, underlining a higher and dose-dependent cvtotoxic effect of oavaverinelohentolamine cornoared to PGEl. Cavemosal tibroblasts are much more resistant when exposed to papaverinelphentolamine than endothelial cells, which may have an importance for the rate of fibrosis during intracavemous pharmacotherapy. I
European
Urology
I
Supplements
3 (2004)
No. 2, pp. 62
21stanbul Istanbul,
The mean serum glucose level, IIEF score and incidences of diabetes coronary artery disease were statistically higher in ED group (p=O.O18; and 0.029 respectively). PPDU assessment resulted in penile arterial in 6 cases, venoocclusive insufficiency in 19 cases and mixed vascular in 5 cases. The distribution of the three eNOS genotypes (eNOS4b/b,
and eNOS4a/a)
between
ED patients
and controls
were
similar
(p>O.O5).
Among the demographic features and risk factors, eNOS4a/b genotype were statistically higher in diabetic patients (p=O.O42). Interestingly, 4 of the 5 patients with severe ED were found to have eNOS4aib genotype. Especially focusing on the determination of the high-risk groups for ED, we failed to find an association between the polymorphism of eNOS gene and ED.
CONCLUSIONS:
On the other hand, we observed
that diabetic
men with
ED have significantly
higher
incidence of eNOS4ti genotype. Considering the multifactorial etiology of ED, further studies with high number of subjects and simultaneous determination of nitric oxide and nitric oxide synthase molecules as well as eNOS genotyping are needed.
SYRIACA
M.3, Kosan M.4, Erdurak
ROOT
EXTRACT
ON
C5, Hafez G.6, Ozgunes
238 A S-HYDROXYTRYPTAMINE PONSE IN RABBIT CAVERNOSAL TO THE ERECTILE PROCESS
BIPHASIC RESMUSCLE: RELEVANCE
0.6, m’,
‘Ankara Numune Education and Research Hospital, 2nd Urology Clinic, Ankara, Turkey, *Faculty of Pharmacy, Ankara University, Department of Pharmacology, Ankara, Turkey, 3Faculty of Pharmacy, Ankara University, Department of Pharmaceutical Botany, Ankara, Turkey, 4Ankara Numune Education and Research Hospital, 32nd Urology Clinic, Ankara, Turkey, 5Faculty of Pharmacy, Ankara University, 2department of Pharmaceutical Botany, Ankara, Turkey, 6Faculty of Pharmacy, Ankara University, ldepartment of Pharmacology, Ankara, Turkey INTRODUCTION & OBJECTIVES: The effects of Fen&go which has been used for its aphrodisiacs properties in Turkey human corpus cavemosum.
(5-HT)-MEDIATED SMOOTH
s)~iaca root extract were investigated on
MATERIAL & METHODS: Strips of human corpus cavernosum were precontracted with phenylephrine (10 mM). Relaxations induced by extract between 1.8- 9.0 mg doses were studied in strips of corpus cavernosum in which tone was elicited by phenylephrine (10 mmoV1). After washing period tissues were incubated with NOsynthase inhibitor, L-NAME or selective soluble guanylate cyclase inhibitor, lH-1.2.4oxadiazolo [4,3-alquinoxalin-lone (ODQ) for 30 min. Furthermore, effects of extract on EFS, acetylcholine, forskolin and SNP- induced relaxation were also evaluated in human corpus cavemosum strips. RESULTS: Extract caused long-lived and dose-dependent relaxations (17.2 + 0.9%, 41.4 i 3.8 %, 60.5 i 3.2%, 86.7+ 2.8% at doses of 1.8, 3.6, 5.4 and 9 mg respectively) in precontracted corporal tissues. The nitric oxide synthesis inhibitor NG-nitro-Larginine methyl ester (L-NAME; 10 mM) produced significant inhibition at the low doses (12.7 + 1.5%, 25.5 & 7.1 at doses of 1.8 and 3.6) although there was no suppression at high doses of extract in the organ bath. The soluble guanylate cyclase inhibitor ODQ (30 mM) significantly affected the extract-induced relaxation (at 9 mg dose of extract, 86.7 +2.8% before, and 35.6 54.95% after ODQ treatment, p
Thompson
C2, Mikhailidis
D.2, Morgan R.3
‘Royal Free Hospital and University College Medical School, Royal Free Campus, Department of Urology and Clinical Biochemistry, Mitcham, United Kingdom, 2Royal Free Hospital and University College Medical School, Royal Free Campus, Department of Clinical Biochemistry, Mitcham, United Kingdom, 3Royal Free Hospital and University College
Medical School, Royal Free Campus, Department of Urology,
Mitcham, United Kingdom
INTRODUCTION & OBJECTIVES: It is of interest that Ketanserin, a S-HT, antagonist and Doxazosin. an aloha-l-blocker which has been shown to have a 5-HT inhihitorv action, were shbwn to ‘have a beneficial effect in the treatment of erectile dysfunction (ED)
(human studies). 5.HT was shown to cause contractions in penile arteries and veins of rat and human, respectively. It is likely that 5-HT might have a similar effect in corpus cavemosal smooth muscle. Therefore, 5-HT might have a role to play on the erectile process as well as the pathogen&s of ED as circulating 5-HT levels are raised in patients with diabetes or peripheral
vascular disease-two
conditions
associated with an increased risk of ED.
MATERIAL & METHODS: The effect of 5-HT on rabbit cavemosal tissues were assessed in organ bath studies. Ketanserin, Doxazosin, Ondansetron and Y-12530 (both 5-HT3 antagonists) were also used to further characterise the receptor profile. Results are presented as median and range (mg/mg=contractionlmg of tissue).
Consistent 5-HT-mediated (10m3M) contractions from baseline recordings were in rabbit cavernosal tissues (n=125). The median contraction was 68.8 mgimg.
RESULTS:
demonstrated
Ketanserin (I Oe5M) completely abolished the contractile response. These contractions were also inhibited by Doxazosin ( 10m4and 10-6M, n=8) in a concentration -dependent manner (1 O4M; 87% reduction, 10-6M; 63.3% reduction). The Doxazosin response was not attributable to
alpha blockade, since alpha-l and 2 antagonists (corynanthine
and yohimbine)
had no effect
on the 5-HT-induced contractions. A biphasic response of transient relaxation followed by substained contraction was observed in 30% of the 5-HT-mediated responses (38 strips of a total of 125). The transient relaxation (median = -20.9 mg/mg; the minus sign = relaxation) was completely abolished by Ondansetron and Y-12530. CONCLUSIONS: 5-HT may have a role to play on the erectile process via its 5-HT,mediated contractile and 5-HT.-mediated relaxant activities. The overall 5.HT induced contractions may represent an in&play between the two S-HT-mediated opposing forces. It is possible that the faster onset 5.HT,-mediated relaxant force modulates the contractile response mediated by activation of the 5-HT,,receptor subtype. The fact that the transient
relaxation is not always present suggests a varlatlon in the rabbit population relating to the erectile 5-HT-mediated response. Our findings further imply that Ketanserin and Doxazosin may be beneficial in the treatment of ED as part of a multi-therapy regime (when a monotherapy failed) via 5-HT dependent mechanisms (rather than exclusively via alpha-l-
adrenergic blockade in the case with Doxazosin).
239 IN VITRO EFFECT OF DIFFERENT DRUGS ON THE VIABILITY ENDOTHELIAL CELLS WA.‘,
INTRACAVERNOUS OF HUMAN
VASOACTIVE CAVERNOSAL
Schultheiss D.‘, Gabouev A.l, Schlote N.l, Wefer J.‘: Mertsching H.Z, Sohn M.3, Stief
240 POLYMORPHISM GENE IN PATIENTS
OF ENDOTHELIAL WITH ERECTILE
Erkan E.‘, Muslumanoglu Kadioglu A.3
A.Y.‘,
Merder
NITRIC OXIDE DYSFUNCTION E.‘, Tefekli
A.‘,
Akbas
SYNTHASE F.2, Ozbek
U.*,
C.G.‘, Jonas U.’
‘Hannover Medical School, Department ofurology, Biotechnology and Artificial Organs, Lebao, Department of Urology, Frankfurt, Germany
Haonover, Germany, 2Leibniz Laboratories for Hannover, Germany, ?St. Markus Hospital,
INTRODUCTION & OBJECTIVES: The endothelium of the corpus cavemosum plays an important role in signal transduction of penile erection. It is directly exposed to intracavemously injected vasoactive agents, which are then passed through the endothelial layer reaching the stromal smooth muscle cells. We investigated commercially available prostaglandin El (Cave+&‘), standard bi-mix of papaverineiphentolamine (AndroskatO) and tri-mix of these agents in physiological dilutions using cultured human cavemosal endothelial cells and fibroblasts. MATERIAL & METHODS: Primary corpus cavemosum endothelial and tibroblast cell cultures were obtained from potent patients (n=12) undergoing penile surgery. 5~10~ cells/well seeded in sixwell plates were exposed for 30min to physiological dilutions of 2Ob.g PGEl (=group P). 30mg papaverineilmg phentolamine (=group A) or t&mix of these agents (=group T) each dissolved in 5m1, lOm1, 2Oml or 5Oml NaCl (= P5 and P50 for the PGEl group etc.). Lactate dehydrogenase (LDH) release as a cytotoxicity marker was measured 6h after drug exposure. The metabolic cell activity was quantified after 48h with the tetrazolium salt-based (MTS) assay and viable cells were counted by performing a Liveil)eada staining. RESULTS: The release of LDH over the first 6h was elevated up to 5-fold (p
‘Haseki Teaching and Research Hospital, Urology, Istanbul, Turkey, University, Medical Faculty, The Institute of Research for Experimental, Turkey, Wanbul University, Medical Faculty, Urology, Istanbul, Turkey
INTRODUCTION & OBJECTIVES: Erectile dysfunction (ED) is a phenomenon which is strictly related to demographic and environmental factors, but the influence of genetic determinants are still not clear. In this trial, we tried to assess the possible role of the polymorphism of endothelial nitric oxide synthase (eNOS) gene on the development and severity of ED. MATERIAL & METHODS: Thirty patients, with a mean age of 56.457.8 (range: 3974) years, who had registered to our andrology policlinic with ED between Febmary 2003 and June 2003 were included in this trial. As the control group, 25 voluntary males, with a mean age of 58.7*9.9 (range: 47-72) years, were included. After a comprehensive physical examination and biochemical evaluation for the risk factors of ED, all cases were searched for the presence of polymorphism of eNOS gene. Detection of the polymorphism was performed in the 4th intron of 7th somatic chromosome which is encoding the eNOS gene, by polymerase chain reaction (PCR). The men with ED were assessed with pharmacopenile dupplex ultrasonography (PPDU) while the potency of the control group was established on the basis of International Index of Erectile Function (IIEF) query. The results of the two groups were compared. For statistical evaluation; chi-square and Student-t tests were used. RESULTS: mellitus and 0.001; 0.023 insufficiency insufficiency eNOS4aib
This effect was less pronounced in the tibroblast cultures where the decrease of viable cells in the more concentrated A and T arouos was less intense after 48h than in endothelial cells (o
European
Urology
I
Supplements
3 (2004)
No. 2, pp. 62
21stanbul Istanbul,
The mean serum glucose level, IIEF score and incidences of diabetes coronary artery disease were statistically higher in ED group (p=O.O18; and 0.029 respectively). PPDU assessment resulted in penile arterial in 6 cases, venoocclusive insufficiency in 19 cases and mixed vascular in 5 cases. The distribution of the three eNOS genotypes (eNOS4b/b,
and eNOS4a/a)
between
ED patients
and controls
were
similar
(p>O.O5).
Among the demographic features and risk factors, eNOS4a/b genotype were statistically higher in diabetic patients (p=O.O42). Interestingly, 4 of the 5 patients with severe ED were found to have eNOS4aib genotype. Especially focusing on the determination of the high-risk groups for ED, we failed to find an association between the polymorphism of eNOS gene and ED.
CONCLUSIONS:
On the other hand, we observed
that diabetic
men with
ED have significantly
higher
incidence of eNOS4ti genotype. Considering the multifactorial etiology of ED, further studies with high number of subjects and simultaneous determination of nitric oxide and nitric oxide synthase molecules as well as eNOS genotyping are needed.