Superoxide Anion Production by NADPH Oxidase Plays a Major Role in Erectile Dysfunction in Middle-Aged Rats: Prevention by Antioxidant Therapy

Fábio H. Silva; Fabíola Z. Mónica; Fernando R. Báu; Ana F. Brugnerotto; Fernanda B.M. Priviero; Haroldo A. Toque; Edson Antunes

doi:10.1111/jsm.12063

Superoxide Anion Production by NADPH Oxidase Plays a Major Role in Erectile Dysfunction in Middle-Aged Rats: Prevention by Antioxidant Therapy

Fábio H. Silva, Fabíola Z. Mónica, Fernando R. Báu, Ana F. Brugnerotto, Fernanda B.M. Priviero, Haroldo A. Toque, Edson Antunes

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41 Scopus citations

Abstract

Introduction.: Prevalence of erectile dysfunction (ED) increases progressively with aging, but the ED pathophysiology at its early stages is still poorly investigated. Aim.: This study aimed to evaluate the functional and molecular alterations of erectile function at middle age, focusing on the contribution of oxidative stress in erectile tissue for the ED. Methods.: Young (3.5-month) and middle-aged (10-month) male Wistar rats were used. Rat corpus cavernosum (RCC) was dissected free and mounted in 10-mL organ baths containing Krebs solution. Intracavernosal pressure (ICP) in anesthetized rats was evaluated. Main Outcome Measures.: Concentration-response curves to endothelium-dependent and endothelium-independent agents, as well as to electrical field stimulation (EFS), were obtained in RCC strips. Measurement of cyclic guanosine monophosphate (cGMP) and expressions of neuronal nitric oxide synthase (nNOS) and endothelial NOS (eNOS), gp91^phox and superoxide dismutase-1 (SOD-1) expressions in RCC were evaluated. Results.: ICP was significantly reduced in middle-aged compared with young rats. RCC relaxations to acetylcholine (10^-8 to 10^-2M), sodium nitroprusside (10^-8 to 10^-2M), sildenafil (10^-9 to 10^-5M), BAY 41-2272 (10^-9 to 10^-5M), and EFS (4-32Hz) were decreased in middle-aged group, which were nearly normalized by apocynin (NADPH oxidase inhibitor; 10^-4M) or SOD (75U/mL). Prolonged treatment with apocynin (85mg/rat/day, 4 weeks) also restored the impaired relaxations in middle-aged rats. Relaxations to 8-bromoguanosine 3′,5′-cyclic monophosphate sodium salt (8-Br-cGMP; 10^-8 to 3×10^-4M) remained unchanged between groups. Basal and stimulated cGMP production were lower in middle-aged group, an effect fully restored by apocynin and SOD. Protein expression of nNOS and phosphorylated eNOS (p-eNOS) (Ser-1177) reduced, whereas gp^91phox mRNA expression increased in RCC from middle-aged rats. Conclusions.: ED in middle-aged rats is associated with decreased NO bioavailability in erectile tissue due to upregulation of NADPH oxidase subunit gp91^phox and downregulation of nNOS/p-eNOS. Antioxidant therapies may be a good pharmacological approach to prevent ED at its early stages.

Original language	English (US)
Pages (from-to)	960-971
Number of pages	12
Journal	Journal of Sexual Medicine
Volume	10
Issue number	4
DOIs	https://doi.org/10.1111/jsm.12063
State	Published - Apr 2013

Keywords

Antioxidant Therapy
Apocynin
Corpus Cavernosum
Erectile Dysfunction
Oxidative Stress
Superoxide Dismutase

ASJC Scopus subject areas

Reproductive Medicine
Obstetrics and Gynecology
Urology

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10.1111/jsm.12063

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@article{2e43e79626fb4ed5bc54d7dba80eb3d2,

title = "Superoxide Anion Production by NADPH Oxidase Plays a Major Role in Erectile Dysfunction in Middle-Aged Rats: Prevention by Antioxidant Therapy",

abstract = "Introduction.: Prevalence of erectile dysfunction (ED) increases progressively with aging, but the ED pathophysiology at its early stages is still poorly investigated. Aim.: This study aimed to evaluate the functional and molecular alterations of erectile function at middle age, focusing on the contribution of oxidative stress in erectile tissue for the ED. Methods.: Young (3.5-month) and middle-aged (10-month) male Wistar rats were used. Rat corpus cavernosum (RCC) was dissected free and mounted in 10-mL organ baths containing Krebs solution. Intracavernosal pressure (ICP) in anesthetized rats was evaluated. Main Outcome Measures.: Concentration-response curves to endothelium-dependent and endothelium-independent agents, as well as to electrical field stimulation (EFS), were obtained in RCC strips. Measurement of cyclic guanosine monophosphate (cGMP) and expressions of neuronal nitric oxide synthase (nNOS) and endothelial NOS (eNOS), gp91phox and superoxide dismutase-1 (SOD-1) expressions in RCC were evaluated. Results.: ICP was significantly reduced in middle-aged compared with young rats. RCC relaxations to acetylcholine (10-8 to 10-2M), sodium nitroprusside (10-8 to 10-2M), sildenafil (10-9 to 10-5M), BAY 41-2272 (10-9 to 10-5M), and EFS (4-32Hz) were decreased in middle-aged group, which were nearly normalized by apocynin (NADPH oxidase inhibitor; 10-4M) or SOD (75U/mL). Prolonged treatment with apocynin (85mg/rat/day, 4 weeks) also restored the impaired relaxations in middle-aged rats. Relaxations to 8-bromoguanosine 3′,5′-cyclic monophosphate sodium salt (8-Br-cGMP; 10-8 to 3×10-4M) remained unchanged between groups. Basal and stimulated cGMP production were lower in middle-aged group, an effect fully restored by apocynin and SOD. Protein expression of nNOS and phosphorylated eNOS (p-eNOS) (Ser-1177) reduced, whereas gp91phox mRNA expression increased in RCC from middle-aged rats. Conclusions.: ED in middle-aged rats is associated with decreased NO bioavailability in erectile tissue due to upregulation of NADPH oxidase subunit gp91phox and downregulation of nNOS/p-eNOS. Antioxidant therapies may be a good pharmacological approach to prevent ED at its early stages.",

keywords = "Antioxidant Therapy, Apocynin, Corpus Cavernosum, Erectile Dysfunction, Oxidative Stress, Superoxide Dismutase",

author = "Silva, {F{\'a}bio H.} and M{\'o}nica, {Fab{\'i}ola Z.} and B{\'a}u, {Fernando R.} and Brugnerotto, {Ana F.} and Priviero, {Fernanda B.M.} and Toque, {Haroldo A.} and Edson Antunes",

note = "Funding Information: F{\'a}bio Henrique da Silva thanks Funda{\c c}{\~a}o de Amparo {\`a} Pesquisa do Estado de S{\~a}o Paulo (FAPESP) for the financial support. ",

year = "2013",

month = apr,

doi = "10.1111/jsm.12063",

language = "English (US)",

volume = "10",

pages = "960--971",

journal = "Journal of Sexual Medicine",

issn = "1743-6095",

publisher = "Wiley-Blackwell",

number = "4",

}

TY - JOUR

T1 - Superoxide Anion Production by NADPH Oxidase Plays a Major Role in Erectile Dysfunction in Middle-Aged Rats

T2 - Prevention by Antioxidant Therapy

AU - Silva, Fábio H.

AU - Mónica, Fabíola Z.

AU - Báu, Fernando R.

AU - Brugnerotto, Ana F.

AU - Priviero, Fernanda B.M.

AU - Toque, Haroldo A.

AU - Antunes, Edson

N1 - Funding Information: Fábio Henrique da Silva thanks Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) for the financial support.

PY - 2013/4

Y1 - 2013/4

N2 - Introduction.: Prevalence of erectile dysfunction (ED) increases progressively with aging, but the ED pathophysiology at its early stages is still poorly investigated. Aim.: This study aimed to evaluate the functional and molecular alterations of erectile function at middle age, focusing on the contribution of oxidative stress in erectile tissue for the ED. Methods.: Young (3.5-month) and middle-aged (10-month) male Wistar rats were used. Rat corpus cavernosum (RCC) was dissected free and mounted in 10-mL organ baths containing Krebs solution. Intracavernosal pressure (ICP) in anesthetized rats was evaluated. Main Outcome Measures.: Concentration-response curves to endothelium-dependent and endothelium-independent agents, as well as to electrical field stimulation (EFS), were obtained in RCC strips. Measurement of cyclic guanosine monophosphate (cGMP) and expressions of neuronal nitric oxide synthase (nNOS) and endothelial NOS (eNOS), gp91phox and superoxide dismutase-1 (SOD-1) expressions in RCC were evaluated. Results.: ICP was significantly reduced in middle-aged compared with young rats. RCC relaxations to acetylcholine (10-8 to 10-2M), sodium nitroprusside (10-8 to 10-2M), sildenafil (10-9 to 10-5M), BAY 41-2272 (10-9 to 10-5M), and EFS (4-32Hz) were decreased in middle-aged group, which were nearly normalized by apocynin (NADPH oxidase inhibitor; 10-4M) or SOD (75U/mL). Prolonged treatment with apocynin (85mg/rat/day, 4 weeks) also restored the impaired relaxations in middle-aged rats. Relaxations to 8-bromoguanosine 3′,5′-cyclic monophosphate sodium salt (8-Br-cGMP; 10-8 to 3×10-4M) remained unchanged between groups. Basal and stimulated cGMP production were lower in middle-aged group, an effect fully restored by apocynin and SOD. Protein expression of nNOS and phosphorylated eNOS (p-eNOS) (Ser-1177) reduced, whereas gp91phox mRNA expression increased in RCC from middle-aged rats. Conclusions.: ED in middle-aged rats is associated with decreased NO bioavailability in erectile tissue due to upregulation of NADPH oxidase subunit gp91phox and downregulation of nNOS/p-eNOS. Antioxidant therapies may be a good pharmacological approach to prevent ED at its early stages.

AB - Introduction.: Prevalence of erectile dysfunction (ED) increases progressively with aging, but the ED pathophysiology at its early stages is still poorly investigated. Aim.: This study aimed to evaluate the functional and molecular alterations of erectile function at middle age, focusing on the contribution of oxidative stress in erectile tissue for the ED. Methods.: Young (3.5-month) and middle-aged (10-month) male Wistar rats were used. Rat corpus cavernosum (RCC) was dissected free and mounted in 10-mL organ baths containing Krebs solution. Intracavernosal pressure (ICP) in anesthetized rats was evaluated. Main Outcome Measures.: Concentration-response curves to endothelium-dependent and endothelium-independent agents, as well as to electrical field stimulation (EFS), were obtained in RCC strips. Measurement of cyclic guanosine monophosphate (cGMP) and expressions of neuronal nitric oxide synthase (nNOS) and endothelial NOS (eNOS), gp91phox and superoxide dismutase-1 (SOD-1) expressions in RCC were evaluated. Results.: ICP was significantly reduced in middle-aged compared with young rats. RCC relaxations to acetylcholine (10-8 to 10-2M), sodium nitroprusside (10-8 to 10-2M), sildenafil (10-9 to 10-5M), BAY 41-2272 (10-9 to 10-5M), and EFS (4-32Hz) were decreased in middle-aged group, which were nearly normalized by apocynin (NADPH oxidase inhibitor; 10-4M) or SOD (75U/mL). Prolonged treatment with apocynin (85mg/rat/day, 4 weeks) also restored the impaired relaxations in middle-aged rats. Relaxations to 8-bromoguanosine 3′,5′-cyclic monophosphate sodium salt (8-Br-cGMP; 10-8 to 3×10-4M) remained unchanged between groups. Basal and stimulated cGMP production were lower in middle-aged group, an effect fully restored by apocynin and SOD. Protein expression of nNOS and phosphorylated eNOS (p-eNOS) (Ser-1177) reduced, whereas gp91phox mRNA expression increased in RCC from middle-aged rats. Conclusions.: ED in middle-aged rats is associated with decreased NO bioavailability in erectile tissue due to upregulation of NADPH oxidase subunit gp91phox and downregulation of nNOS/p-eNOS. Antioxidant therapies may be a good pharmacological approach to prevent ED at its early stages.

KW - Antioxidant Therapy

KW - Apocynin

KW - Corpus Cavernosum

KW - Erectile Dysfunction

KW - Oxidative Stress

KW - Superoxide Dismutase

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DO - 10.1111/jsm.12063

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AN - SCOPUS:84875678316

SN - 1743-6095

VL - 10

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JO - Journal of Sexual Medicine

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ER -

Superoxide Anion Production by NADPH Oxidase Plays a Major Role in Erectile Dysfunction in Middle-Aged Rats: Prevention by Antioxidant Therapy

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Keywords

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