Additive effects of the Rho kinase inhibitor Y-27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure

Pieter Uvin, Maarten Albersen, Ine Bollen, Maarten Falter, Emmanuel Weyne, Loes Linsen, Hanna Tinel, Peter Sandner, Trinity J. Bivalacqua, Dirk J M K De Ridder, Frank Van der Aa, Bert Brône, Koenraad Van Renterghem

Research output: Contribution to journalArticle

Abstract

Objectives: To evaluate the expression of the Rho/Rho-associated protein kinase (ROCK) pathway in the corpus cavernosum of patients with severe erectile dysfunction (ED) compared with healthy human corpus cavernosum, and to test the functional effects of two Rho kinase inhibitors (RKIs) on erectile tissue of patients with severe ED, which did not respond to phosphodiesterase type 5 inhibitors (PDE5Is). Patients and Methods: Human corpus cavernosum samples were obtained after consent from men undergoing penile prosthesis implantation (n = 7 for organ bath experiments, n = 17 for quantitative PCR [qPCR]). Potent control subjects (n = 5) underwent penile needle biopsy. qPCR was performed for the expression of RhoA and ROCK subtypes 1 and 2. Immunohistochemistry staining against ROCK and α smooth muscle actin (αSMA) was performed on the corpus cavernosum of patients with ED. Tissue strips were precontracted with phenylephrine and incubated with 1 μm of the PDE5I vardenafil or with DMSO (control). Subsequently, increasing concentrations of the RKIs azaindole or Y-27632 were added, and relaxation of tissue was quantified. Results: The expression of ROCK1 was unchanged (P > 0.05), while ROCK2 (P < 0.05) was significantly upregulated in patients with ED compared with controls. ROCK1 and ROCK2 protein colocalized with αSMA, confirming the presence of this kinase in cavernous smooth muscle cells and/or myofibroblasts. After incubation with DMSO, 10 μm azaindole and 10 μm Y-27632 relaxed precontracted tissues with 49.5 ± 7.42% (P = 0.1470 when compared with vehicle) and 85.9 ± 10.3% (P = 0.0016 when compared with vehicle), respectively. Additive effects on relaxation of human corpus cavernosum were seen after preincubation with 1 μm vardenafil. Conclusion: The RKI Y-27632 causes a significant relaxation of corpus cavernosum in tissue strips of patients with severe ED. The additive effect of vardenafil and Y-27632 shows that a combined inhibition of Rho-kinase and phosphodiesterase type 5 could be a promising orally administered treatment for severe ED.

Original languageEnglish (US)
Pages (from-to)325-332
Number of pages8
JournalBJU International
Volume119
Issue number2
DOIs
StatePublished - Feb 1 2017

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Phosphodiesterase 5 Inhibitors
rho-Associated Kinases
Erectile Dysfunction
Dimethyl Sulfoxide
Smooth Muscle
Actins
Polymerase Chain Reaction
Penile Implantation
Type 5 Cyclic Nucleotide Phosphodiesterases
Myofibroblasts
Needle Biopsy
Phenylephrine
Protein Kinases
Phosphotransferases
Immunohistochemistry
Staining and Labeling
Proteins
Inhibition (Psychology)

Keywords

  • corpus cavernosum
  • erectile dysfunction
  • phosphodiesterase type 5 inhibitor
  • Rho kinase inhibitor
  • Rho/Rho associated protein kinase (ROCK) pathway
  • Y-27632

ASJC Scopus subject areas

  • Urology

Cite this

Additive effects of the Rho kinase inhibitor Y-27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure. / Uvin, Pieter; Albersen, Maarten; Bollen, Ine; Falter, Maarten; Weyne, Emmanuel; Linsen, Loes; Tinel, Hanna; Sandner, Peter; Bivalacqua, Trinity J.; De Ridder, Dirk J M K; Van der Aa, Frank; Brône, Bert; Van Renterghem, Koenraad.

In: BJU International, Vol. 119, No. 2, 01.02.2017, p. 325-332.

Research output: Contribution to journalArticle

Uvin, P, Albersen, M, Bollen, I, Falter, M, Weyne, E, Linsen, L, Tinel, H, Sandner, P, Bivalacqua, TJ, De Ridder, DJMK, Van der Aa, F, Brône, B & Van Renterghem, K 2017, 'Additive effects of the Rho kinase inhibitor Y-27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure' BJU International, vol 119, no. 2, pp. 325-332. DOI: 10.1111/bju.13691

Uvin, Pieter; Albersen, Maarten; Bollen, Ine; Falter, Maarten; Weyne, Emmanuel; Linsen, Loes; Tinel, Hanna; Sandner, Peter; Bivalacqua, Trinity J.; De Ridder, Dirk J M K; Van der Aa, Frank; Brône, Bert; Van Renterghem, Koenraad / Additive effects of the Rho kinase inhibitor Y-27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure.

In: BJU International, Vol. 119, No. 2, 01.02.2017, p. 325-332.

Research output: Contribution to journalArticle

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title = "Additive effects of the Rho kinase inhibitor Y-27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure",
keywords = "corpus cavernosum, erectile dysfunction, phosphodiesterase type 5 inhibitor, Rho kinase inhibitor, Rho/Rho associated protein kinase (ROCK) pathway, Y-27632",
author = "Pieter Uvin and Maarten Albersen and Ine Bollen and Maarten Falter and Emmanuel Weyne and Loes Linsen and Hanna Tinel and Peter Sandner and Bivalacqua, {Trinity J.} and {De Ridder}, {Dirk J M K} and {Van der Aa}, Frank and Bert Brône and {Van Renterghem}, Koenraad",
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T1 - Additive effects of the Rho kinase inhibitor Y-27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure

AU - Uvin,Pieter

AU - Albersen,Maarten

AU - Bollen,Ine

AU - Falter,Maarten

AU - Weyne,Emmanuel

AU - Linsen,Loes

AU - Tinel,Hanna

AU - Sandner,Peter

AU - Bivalacqua,Trinity J.

AU - De Ridder,Dirk J M K

AU - Van der Aa,Frank

AU - Brône,Bert

AU - Van Renterghem,Koenraad

PY - 2017/2/1

Y1 - 2017/2/1

N2 - Objectives: To evaluate the expression of the Rho/Rho-associated protein kinase (ROCK) pathway in the corpus cavernosum of patients with severe erectile dysfunction (ED) compared with healthy human corpus cavernosum, and to test the functional effects of two Rho kinase inhibitors (RKIs) on erectile tissue of patients with severe ED, which did not respond to phosphodiesterase type 5 inhibitors (PDE5Is). Patients and Methods: Human corpus cavernosum samples were obtained after consent from men undergoing penile prosthesis implantation (n = 7 for organ bath experiments, n = 17 for quantitative PCR [qPCR]). Potent control subjects (n = 5) underwent penile needle biopsy. qPCR was performed for the expression of RhoA and ROCK subtypes 1 and 2. Immunohistochemistry staining against ROCK and α smooth muscle actin (αSMA) was performed on the corpus cavernosum of patients with ED. Tissue strips were precontracted with phenylephrine and incubated with 1 μm of the PDE5I vardenafil or with DMSO (control). Subsequently, increasing concentrations of the RKIs azaindole or Y-27632 were added, and relaxation of tissue was quantified. Results: The expression of ROCK1 was unchanged (P > 0.05), while ROCK2 (P < 0.05) was significantly upregulated in patients with ED compared with controls. ROCK1 and ROCK2 protein colocalized with αSMA, confirming the presence of this kinase in cavernous smooth muscle cells and/or myofibroblasts. After incubation with DMSO, 10 μm azaindole and 10 μm Y-27632 relaxed precontracted tissues with 49.5 ± 7.42% (P = 0.1470 when compared with vehicle) and 85.9 ± 10.3% (P = 0.0016 when compared with vehicle), respectively. Additive effects on relaxation of human corpus cavernosum were seen after preincubation with 1 μm vardenafil. Conclusion: The RKI Y-27632 causes a significant relaxation of corpus cavernosum in tissue strips of patients with severe ED. The additive effect of vardenafil and Y-27632 shows that a combined inhibition of Rho-kinase and phosphodiesterase type 5 could be a promising orally administered treatment for severe ED.

AB - Objectives: To evaluate the expression of the Rho/Rho-associated protein kinase (ROCK) pathway in the corpus cavernosum of patients with severe erectile dysfunction (ED) compared with healthy human corpus cavernosum, and to test the functional effects of two Rho kinase inhibitors (RKIs) on erectile tissue of patients with severe ED, which did not respond to phosphodiesterase type 5 inhibitors (PDE5Is). Patients and Methods: Human corpus cavernosum samples were obtained after consent from men undergoing penile prosthesis implantation (n = 7 for organ bath experiments, n = 17 for quantitative PCR [qPCR]). Potent control subjects (n = 5) underwent penile needle biopsy. qPCR was performed for the expression of RhoA and ROCK subtypes 1 and 2. Immunohistochemistry staining against ROCK and α smooth muscle actin (αSMA) was performed on the corpus cavernosum of patients with ED. Tissue strips were precontracted with phenylephrine and incubated with 1 μm of the PDE5I vardenafil or with DMSO (control). Subsequently, increasing concentrations of the RKIs azaindole or Y-27632 were added, and relaxation of tissue was quantified. Results: The expression of ROCK1 was unchanged (P > 0.05), while ROCK2 (P < 0.05) was significantly upregulated in patients with ED compared with controls. ROCK1 and ROCK2 protein colocalized with αSMA, confirming the presence of this kinase in cavernous smooth muscle cells and/or myofibroblasts. After incubation with DMSO, 10 μm azaindole and 10 μm Y-27632 relaxed precontracted tissues with 49.5 ± 7.42% (P = 0.1470 when compared with vehicle) and 85.9 ± 10.3% (P = 0.0016 when compared with vehicle), respectively. Additive effects on relaxation of human corpus cavernosum were seen after preincubation with 1 μm vardenafil. Conclusion: The RKI Y-27632 causes a significant relaxation of corpus cavernosum in tissue strips of patients with severe ED. The additive effect of vardenafil and Y-27632 shows that a combined inhibition of Rho-kinase and phosphodiesterase type 5 could be a promising orally administered treatment for severe ED.

KW - corpus cavernosum

KW - erectile dysfunction

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KW - Rho kinase inhibitor

KW - Rho/Rho associated protein kinase (ROCK) pathway

KW - Y-27632

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