STIMULATION OF THE CYCLIC AMP/GMP SIGNALLING ENHANCES THE RELAXATION OF ISOLATED HUMAN DETRUSOR SMOOTH MUSCLE ACHIEVED BY PHOSPHODIESTERASE INHIBITORS.

Phosphodiesterase (PDE) enzymes are considered being key proteins in controlling the function of smooth musculature in the human urinary tract. The use of PDE inhibitors (PDE-Is) to treat erectile dysfunction and lower urinary tract symptomatology (LUTS) secondary to benign prostatic hyperplasia (BPH) is well established. It has been shown that PDE-Is can reverse the tension induced by means of muscarinergic agents of detrusor smooth muscle and enhance the production of cyclic nucleotides. In clinical settings, the PDE1 inhibitor vinpocetine had beneficial effects in patients presenting with voiding dysfunctions. This prompted us to evaluate further the mechanism of action of PDE-Is on bladder smooth musculature. Using the tissue bath technique, relaxant responses of human detrusor smooth muscle, challenged by acetylcholine (1 µM), to vinpocetine (PDE1-I), rolipram (PDE4-I), MY 5445 and sildenafil (PDE5-Is) (0.1 µM, 1 µM, and 10 µM) were investigated with and without pre-exposure of the tissue to threshold concentrations of the NO donor drug sodium nitroprusside (SNP) or adenylyl cyclase activator forskolin (0.02 µM). The non-specific PDE-I papaverine was used as a reference compound. The cumulative addition of forskolin or SNP exerted a pronounced reversion of the tension induced by means of ACh, starting at a concentration of 1 µM (forskolin, -25,6%) and 0.1 µM (SNP, -20%), respectively. There were marginal responses of the detrusor smooth musculature to the PDE-Is, the relaxation measured ranged from -12% (vinpocetine/sildenafil) to -19% (rolipram, MY 5445). Exposure of the tissue to a threshold concentration of SNP increased the reversion of tension induced by vinpocetine (-40%), rolipram (-50%) and MY 5445 (-45%). An enhancement in the potency of the drugs was also registered. A threshold concentration of SNP did not significantly affect the maximum reversion of tension brought about by sildenafil but added positively to the in vitro potency of the PDE5-I. PDE inhibitors may tend to be more effective in systems characterized by an enhanced production of cyclic AMP/GMP (such as urogenital tissues in vivo). Our findings may explain how PDE inhibitors can affect symptoms of the overactive bladder.

Endopeptidase inhibition attenuates the contraction induced by big endothelin-1 of isolated human penile erectile tissue.

Peptides, such as C-type natriuretic peptide (CNP), vasoactive intestinal polypeptide (VIP) and endothelin 1 (ET-1), are involved in the control of penile erectile tissue (corpus cavernosum = CC). Inhibiting the degradation of CNP and VIP or conversion of Big ET-1 into ET-1 by endopeptidase enzymes should result in an enhancement of CC smooth muscle relaxation. Using the tissue bath technique, responses of isolated CC, challenged by noradrenaline (NA, 1 µm), to increasing concentrations of the endopeptidase inhibitor KC 12615 (1 nm - 10 µm), CNP and VIP (0.1 nm - 1 µm), were investigated. Effects of CNP, VIP and Big ET-1 (0.1 nm - 100 nm) on the tissue tension were also evaluated following pre-exposure to 10 µm of KC 12615. Big ET-1 induced contraction of the CC amounting to a force generation of 1,200 mg. The contraction was attenuated in the presence of KC 12615 by 35% and 50%, respectively. The tension induced by NA was reversed by VIP and CNP to 38.7% ± 15.8% and 61% ± 13%, respectively, of the initial force. The findings might be of significance with regard to future pharmacological treatment options for male ED, where an endothelial dysfunction exists.

Effects of endopeptidase inhibition on the relaxation response of isolated human penile erectile tissue to vasoactive peptides.

Peptides, such as CNP, CGRP and VIP, are involved in the function of male penile erectile tissue. Tissue levels of said peptides are controlled by the endopeptidase enzymes. Theoretically, the inhibition of the degradation of CNP, CGRP and/or VIP should result in an enhancement in penile smooth muscle relaxation. The effects were investigated of CNP or VIP (0.1 nm-1 µm), without and following pre-exposure of the tissue to a threshold concentration of the endopeptidase inhibitor KC 12615 (10 µm, for 20 min), on the reversion of tension induced by means of electrical field stimulation. Drug effects on the production of cyclic AMP/GMP were also evaluated. Neither KC 12615, CNP and VIP nor the combination of CNP plus KC 12615 or VIP plus KC 12615 increased the response of the tissue to EFS. While no effects were observed of a pre-exposure of the tissue to KC 12615 on the production of cyclic AMP in the presence of VIP, an enhancement was registered in the accumulation of cyclic AMP in the presence of CNP plus KC 12615. Further studies are indicated to investigate whether endopeptidase inhibitors might tend to be more effective in tissues affected by a decreased local production of vasoactive peptides.