Vardenafil modulates bladder contractility through cGMP-mediated inhibition of RhoA/Rho kinase signaling pathway in spontaneously hypertensive rats.
BACKGROUND
Phosphodiesterase type 5 inhibitors (PDE5i), the most widely used drugs for erectile dysfunction, could also improve lower urinary tract symptoms, essentially due to overactive bladder (OAB), a condition hypothesized to be a result of an increased RhoA/Rho-kinase (ROCK) signaling. Phosphorylation/inactivation of RhoA by cyclic guanosine monophosphate (cGMP)-dependent protein kinase (PKG) activity has been described in vascular smooth muscle.


OBJECTIVE
The aim of this paper was to investigate whether vardenafil-induced cGMP accumulation reduces RhoA/ROCK signaling in bladder.


METHODS
Spontaneously hypertensive rats (SHRs), a strain genetically prone to develop OAB, were treated with vardenafil (10 mg/kg/day) for 2 weeks. Wistar-Kyoto rats (WKY) were used as control. In vitro experiments were performed in human bladder smooth muscle cells (hBCs).


METHODS
Urodynamic parameters were registered in vivo in anesthetized WKY and SHRs. RhoA/ROCK activity in bladder was evaluated by molecular and functional studies in tissues and cells.


RESULTS
The intercontraction interval and bladder capacity, and were decreased in SHRs and restored by vardenafil. The in vitro relaxant effect of the ROCK inhibitor Y-27632 was higher in bladder strips from SHR than from WKY and reduced by vardenafil. Nomega-nitro-L-arginine-methyl-ester (a NO-synthase inhibitor, 40 mg/kg/day during the last week of the 2-week treatment with vardenafil) partially antagonized vardenafil effect on Y-27632 responsiveness. Vardenafil prevented RhoA membrane translocation/activation, decreased ROCK activity, and increased cGMP levels in vivo (rat) and in vitro (hBCs). Exposing hBCs to vardenafil increased Ser(188) RhoA phosphorylation, to the same extent as the PDE5-insensitive PKG agonist Sp-8-Br-PET-cGMP. Moreover, vardenafil inhibited several RhoA-dependent functions in hBCs, including smooth muscle gene transcription and endothelin-1-induced migration. These effects were reverted by the PKG inhibitor KT 5823, further suggesting a cGMP/PKG-dependency. In hBCs, vardenafil was active in the low nanomolar range.


CONCLUSIONS
This is the first study demonstrating that the effect of vardenafil on OAB could be partially determined by a cGMP-dependent RhoA/ROCK signaling inhibition.
Morelli
Annamaria
A
Department of Clinical Physiopathology, University of Florence, Florence, Italy.
Filippi
Sandra
S
Department of Clinical Physiopathology, University of Florence, Florence, Italy.
Sandner
Peter
P
Bayer Healthcare, Wuppertal, Germany.
Fibbi
Benedetta
B
Department of Clinical Physiopathology, University of Florence, Florence, Italy.
Chavalmane
Aravinda K
AK
Department of Clinical Physiopathology, University of Florence, Florence, Italy.
Silvestrini
Enrico
E
Department of Clinical Physiopathology, University of Florence, Florence, Italy.
Sarchielli
Erica
E
Department of Anatomy, Histology and Forensic Medicine, University of Florence, Florence, Italy.
Vignozzi
Linda
L
Department of Clinical Physiopathology, University of Florence, Florence, Italy.
Gacci
Mauro
M
Department of Urology, University of Florence, Florence, Italy.
Carini
Marco
M
Department of Urology, University of Florence, Florence, Italy.
Vannelli
Gabriella B
GB
Department of Anatomy, Histology and Forensic Medicine, University of Florence, Florence, Italy.
Maggi
Mario
M
Department of Clinical Physiopathology, University of Florence, Florence, Italy;. Electronic address: m.maggi@DFC.UNIFI.IT.
D000818:Animals; D015153:Blotting, Western; D006152:Cyclic GMP / Q000378:metabolism*; D006973:Hypertension / Q000201:enzymology* / Q000503:physiopathology*; D007093:Imidazoles / Q000494:pharmacology*; D009119:Muscle Contraction / Q000187:drug effects*; D009130:Muscle, Smooth / Q000187:drug effects / Q000201:enzymology; D058986:Phosphodiesterase 5 Inhibitors; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D010879:Piperazines / Q000494:pharmacology*; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects*; D013450:Sulfones / Q000494:pharmacology; D014227:Triazines / Q000494:pharmacology; D053201:Urinary Bladder, Overactive / Q000188:drug therapy*; D014563:Urodynamics / Q000187:drug effects; D000069058:Vardenafil Dihydrochloride; D054460:rho-Associated Kinases / Q000187:drug effects*; D020742:rhoA GTP-Binding Protein / Q000187:drug effects*
D007093:Imidazoles; D058986:Phosphodiesterase 5 Inhibitors; D010726:Phosphodiesterase Inhibitors; D010879:Piperazines; D013450:Sulfones; D014227:Triazines; D000069058:Vardenafil Dihydrochloride; D054460:rho-Associated Kinases; D020742:rhoA GTP-Binding Protein; D006152:Cyclic GMP