Corporal Smooth Muscle Cells Research Articles

Gene Transfer of TRPC6DN (Dominant Negative) Restores Erectile Function in Diabetic Rats

Transient receptor potential (TRP) channels play an important role in modulating intracellular Ca(2+) ([Ca(2+)](i)) levels. We examined the hypothesis that overexpression of TRPC6(DN) (dominant negative) may contribute to decreased [Ca(2+)](i) levels in corporal smooth muscle (CSM). We also investigated whether gene transfer of TRPC6(DN) could restore erectile function in diabetic rats. For the in vitro study, the K(Ca), K(ATP), and TRPC6(DN) channel genes were transferred using cDNA, into cultured human CSM cells and human embryonic kidney cells. For the in vivo study, young adult rats were divided into three groups: normal controls; diabetic controls transfected with vector only; and a diabetic group transfected with pcDNA of the TRPC6(DN) gene. After gene transfer, the effects of reducing [Ca(2+)](i) levels were assessed by Fura-2-based imaging analysis. The intracavernosal pressure (ICP) response to cavernosal nerve stimulation was assessed after intracorporal injection of TRPC6(DN) pcDNA. The transgene expression of the TRPC6(DN) was examined by reverse transcription polymerase chain reaction (RT-PCR) in rats transfected with TRPC6(DN) pcDNA. Gene transfer of ion channels effectively reduced [Ca(2+)](i). Among these channels, transfer of the TRPC6(DN) gene resulted in the greatest reduction of [Ca(2+)](i) in human CSM. The mean (+/-standard error of the mean) ratio of ICP to mean arterial pressure (BP) in the gene-transfer rats was 79.4 +/- 2.4% (N = 8). This was significantly higher than that in control rats (55.6 +/- 3.7% [N = 8]), and similar to that in the young control rats (83 +/- 2.2% [N = 12]). The RT-PCR showed expression of TRPC6(DN) genes in the transfected rats. Gene transfer of TRPC6(DN) not only reduced [Ca(2+)](i) in human CSM but also restored erectile function in diabetic rats. These results suggest that pcDNA transfer of TRPC6(DN) may represent a promising new form of therapy for the treatment of male erectile dysfunction in the future.

Evaluation of the biocompatibility and mechanical properties of naturally derived and synthetic scaffolds for urethral reconstruction

The aim of this study was to evaluate the mechanical properties and biocompatibility of biomaterials, including bladder submucosa (BAMG), small intestinal submucosa (SIS), acellular corpus spongiosum matrix (ACSM), and polyglycolic acid (PGA), to identify the optimal scaffold for urethral tissue engineering. Tensile mechanical testing was conducted to evaluate mechanical properties of each scaffold. Rabbit corporal smooth muscle cells were cultured with the extracts of biomaterials and mitochondrial metabolic activity assay was used to determine the cytotoxicity of scaffold. The pore sizes of each scaffold were measured. Additionally, smooth muscle cells were seeded on biomaterials. Cell infiltration was evaluated. Mechanical evaluation showed that Young modulus, stress at break in ACSM were prior to those in other biomaterials (p < 0.05). MTT assay confirmed that all scaffolds supported normal cellular mitochondrial metabolic without inducing cytotoxic events. SEM demonstrated that PGA has the largest pore size (>200 microm). The ACSM has different pore sizes in urethral (<5 microm) and cavernosal surfaces (>10 microm). Widespread distribution of cells could be observed in PGA 14 days after seeding. Multilayer cellular coverage developed in BAMG and urethral surface of ACSM without any sign of cellular invasion. Moderated cellular penetration could be found in SIS and cavernosal surface of ACSM. Although each scaffold demonstrated suitable mechanical properties, which is similar to normal urethra, ACSM showed better response in some parameters than those in other biomaterials. It suggested that this scaffold may be an alternative for urethral reconstruction in the future. (c) 2010 Wiley Periodicals, Inc. J Biomed Mater Res, 2010.

Early onset of fibrosis within the arterial media in a rat model of type 2 diabetes mellitus with erectile dysfunction

To determine, in the obese Zucker fa/fa rat (OZR), whether the loss in smooth muscle cells (SMCs) as well as the increase in fibrosis that occurs within the corpora cavernosa accompanying corporal veno-occlusive dysfunction (CVOD), also occurs within the media of the arterial tree. The penis and aorta from both 7-month-old male diabetic OZR (5 months of diabetes) and aged-matched nondiabetic lean Zucker rats (LZR) rats were harvested (eight per group). The penis and aorta were subjected to histo- or immnohistochemistry, followed by quantitative image analysis (QIA) to determine the contents of SMC, collagen and the pro-fibrotic transforming growth factor (TGF)beta1. The turnover of SMCs was assessed by terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labelling (TUNEL) and proliferating cell nuclear antigen (PCNA) assays. Quantitative Western blots determined calponin (SMC marker) and PCNA, and hydroxyproline was used for collagen. In vitro relaxation of corporal strips was measured. In vitro relaxation of corporal tissue from OZR was considerably less than in the LZR. In the media of the penile dorsal artery (PDA) of OZR, there was a considerable reduction in the SMC content and the SMC/collagen ratio, as well as an increase in apoptosis, but there were no changes in PCNA or TGFbeta1 expression, or in the intima-media/lumen ratio. In the aorta of the OZR, in contrast to the PDA, there was a reduction in PCNA as well as a more pronounced decrease in the SMC/collagen ratio, mainly from an increase in collagen, but there were no changes in TGFbeta1 or the wall/lumen morphometry. In the OZR, Western blots of aortic tissue confirmed the decrease in PCNA and a reduction in the SMC marker calponin. These data show that 5 months after the onset of hyperglycaemia in the OZR, the rats develop both abnormal corporal SMC relaxation and a generalized fibrosis of the arterial media of both the large and small diameter vessels. It is possible that this pan-fibrosis of the media of the arterial system might contribute to the diabetes-related ED that occurs during this period in this rat model.

Fibrosis and Loss of Smooth Muscle in the Corpora Cavernosa Precede Corporal Veno-Occlusive Dysfunction (CVOD) Induced by Experimental Cavernosal Nerve Damage in the Rat

Corporal veno-occlusive dysfunction (CVOD), which usually is associated with a loss of smooth muscle cells (SMC) and an increase in fibrosis within the corpora cavernosa, can be induced by an injury to the cavernosal nerves. The corporal tissue expresses inducible nitric oxide synthase (iNOS), presumably as an antifibrotic and SMC-protective response. We studied the temporal relationship in the corpora between the expression of iNOS, other histological and biochemical changes, and the development of CVOD, after bilateral cavernosal nerve resection (BCNR) in the rat. Rats underwent either BCNR or sham operation. Cavernosometry was performed 1, 3, 7, 15, 30, and 45 days (N = 8/groups) after surgery. Penile tissue sections were subjected to Masson trichrome staining for SMC and collagen, and immunodetection for alpha smooth muscle actin, iNOS, neuronal NOS (nNOS), endothelial NOS (eNOS), proliferating cell nuclear antigen (PCNA), and terminal transferase dUTP nick end labeling (TUNEL). Quantitative western blot analysis was done in homogenates. Time course on the development of fibrosis and CVOD. Following BCNR, CVOD was detectable 30 days later, and it became more pronounced by 45 days. In contrast, the SMC/collagen ratio in the BCNR corpora was reduced at 7 days and bottomed at 30 and 45 days, due in part to the reduction of SMC, presumably caused by an increase in apoptosis peaking at 3 days. PCNA also peaked at 3 days, but then decayed. nNOS was reduced early (3-7 days) and disappeared at 30 days, whereas eNOS was not affected. iNOS was induced at day 3, and steadily increased peaking at 30 days. CVOD develops in the BCNR rat as a result of the early loss of corporal SMC by the neuropraxia-induced apoptosis, which the initial cell replication response cannot counteract, followed by fibrosis. The time course of iNOS induction supports the antifibrotic role of iNOS.

Sildenafil Inhibits Superoxide Formation and Prevents Endothelial Dysfunction in a Mouse Model of Secondhand Smoke Induced Erectile Dysfunction

We determined the effect of passive secondhand cigarette smoke on 1) erectile function in vivo, 2) molecular mechanisms involved in penile vascular function, and 3) erectile function and penile molecular signaling in the presence of phosphodiesterase type 5 inhibitor therapy. Four groups of mice were used, including group 1--controls, group 2--mice exposed to 3 weeks of secondhand smoke (5 hours per day for 5 days per week), group 3--control plus sildenafil (100 mg/kg per day) and group 4--smoke exposed plus sildenafil (100 mg/kg per day). Cavernous nerve electrical stimulation and intracavernous injection of acetylcholine were done to assess erectile function. Constitutive and inducible nitric oxide synthase activity, reactive oxygen species generation, nitrotyrosine formation and superoxide anion levels were assessed. Decreased erectile responses to cavernous nerve electrical stimulation and impaired endothelium dependent erectile responses to ACh in mice exposed to secondhand smoke were observed. Superoxide anion was increased in endothelial and corporeal smooth muscle cells of smoking mouse penises. In mice exposed to secondhand smoke constitutive nitric oxide synthase activity was decreased, and inducible nitric oxide synthase activity, reactive oxygen species generation and nitrotyrosine formation increased. Sildenafil therapy restored constitutive nitric oxide synthase activity in the penis of smoking mice, decreased inducible nitric oxide synthase activity, reactive oxygen species generation and nitrotyrosine formation, and improved erectile responses to cavernous nerve electrical stimulation and acetylcholine. Short-term exposure to secondhand smoke impairs erectile function through excessive penile reactive oxygen species signaling and inducible nitric oxide synthase activity. Decreased penile constitutive nitric oxide synthase activity may be attributable to the decreased endothelial nitric oxide synthase activity resulting from increased oxidative stress. Sildenafil therapy restored nitric oxide synthase activity and decreased reactive oxygen species signaling, resulting in improved erectile function.

Transcription of G-Protein Coupled Receptors in Corporeal Smooth Muscle is Regulated by the Endogenous Neutral Endopeptidase Inhibitor Sialorphin

Several reports suggest that the rat Vcsa1 gene is down-regulated in models of erectile dysfunction. The Vcsa protein product sialorphin is an endogenous neutral endopeptidase inhibitor and its down-regulation could result in prolonged activation of G-protein activated signaling pathways by their peptide agonists. We investigated whether Vcsa1 down-regulation could result in an adaptive change in GPCR (G-protein coupled receptor) expression. Gene expression in cultured rat corporeal smooth muscle cells following treatment with siRNA directed against Vcsa1 or the neutral endopeptidase gene was analyzed using microarray and quantitative reverse transcriptase-polymerase chain reaction. In rats Vcsa1 is one of the most down-regulated genes following bilateral transection of the cavernous nerves. In that animal model we also investigated whether Vcsa1 down-regulation was accompanied by similar changes in gene expression in corporeal smooth muscle cells in which Vcsa1 was knocked down in vitro. Microarray analysis and quantitative reverse transcriptase-polymerase chain reaction demonstrated that corporeal smooth muscle cells treated in vitro with siRNA against Vcsa1 resulted in GPCR up-regulation as a functional group. In contrast, treatment of corporeal smooth muscle cells that lowered neutral endopeptidase activity resulted in decreased GPCR expression. These results suggest that the peptide product of Vcsa1, sialorphin, can effect GPCR expression by acting on neutral endopeptidase. In animals with bilaterally transected cavernous nerves the decreased Vcsa1 expression is accompanied by increased GPCR expression in cavernous tissue. These experiments suggest that the mechanism by which Vcsa1 modulates erectile function is partly mediated through changes in GPCR expression.

Effect of Testosterone on Potassium Channel Opening in Human Corporal Smooth Muscle Cells

In humans, the role of testosterone in sexual functions, including sexual desire, nocturnal penile erections, and ejaculatory volume, has been relatively well established. However, the effects of testosterone on intrapenile structure in humans remains controversial. We assessed the direct effects of testosterone on potassium channels in human corporal smooth muscle cells, in an effort to understand the mechanisms inherent to the testosterone-induced relaxation of corporal smooth muscle cells at the cellular and molecular levels. Methods. We conducted electrophysiologic studies using cultured human corporal smooth muscle cells. We evaluated the effects of testosterone on potassium channels-BK(Ca) and K(ATP) channels-by determining the whole-cell currents and single-channel activities. For the electrophysiologic recordings, whole-cell and cell-attached configuration patch-clamp techniques were utilized. Changes in whole-cell currents and channel activities of BK(Ca) and K(ATP) channels by testosterone. Results. Testosterone (200 nM) significantly increased the single-channel activity of calcium-activated potassium (BK(Ca)) channels and whole-cell K(+) currents by 443.4 +/- 83.4% (at +60 mV; N = 11, P < 0.05), and this effect was abolished by tetraethylammonium (TEA) (1 mM), a BK(Ca) channel blocker. The whole-cell inward K(+) currents of the K(ATP) channels were also increased by 226.5 +/- 49.3% (at -100 mV; N = 7, P < 0.05). In the presence of a combination of vardenafil (10 nM) and testosterone (200 nM), the BK(Ca) channel was activated to a significantly higher degree than was induced by testosterone alone. The results of patch-clamp studies provided direct molecular evidence that testosterone stimulates the activity of BK(Ca) channels and K(ATP) channels. An understanding of the signaling mechanisms that couple testosterone receptor activation to potassium channel stimulation will provide us with an insight into the cellular processes underlying the vasorelaxant effects of testosterone.

Diabetes‐related alterations in second messenger regulation of potassium channel activity in corporal smooth muscle cells

Corporal smooth muscle (CSM) relaxation mediates penile erection, and second messenger activation of the KCa and KATP channel subtypes plays a major role in this process. The goal of this study was to evaluate the potential role of altered second messenger activated hyperpolarizing K+ currents in CSM to diabetes (DM)‐related erectile dysfunction (ED). Corporal tissue was excised from the STZ‐DM rats following 2 months of DM. Whole cell currents were recorded from short‐term cultured CSM cells of DM and age‐matched control (AMC) rats; in the absence and presence of either cAMP (prostaglandin E1 (PGE1; 10 uM) or 8‐Br‐cAMP (1 mM)) or cGMP (sodium nitroprusside (SNP; 10 uM) or 8‐Br‐cGMP (1 mM)) activation. Holding potential was −70 mV, with I‐V curves constructed in 10 mV increments from −60 to +100 mV. Resting potentials were lower in DM (−44 ± 1.3 mV; n=40) than AMC (−56 ± 1.4 mV; n=40) (p&lt;0.05). Outward whole cell K+ currents were reduced in DM (206 ± 11 pA; n=40) relative to AMC (321 ± 16 pA; n=40) (p&lt;0.05). K+ currents induced by cGMP, SNP, cAMP and PGE1 were significantly diminished in DM relative to AMC. IBTX (100 nM) decreased cGMP‐activated K+ currents more than glibenclamide (GLIB; 10 uM) in AMC rats, but less than GLIB in DM rats. The data suggest that cGMP and cAMP‐mediated K+ currents are decreased in DM rats, and that altered regulation of the KCa and KATP channel subtypes might contribute to DM‐related ED in rats.

The effects of dopamine receptor agonists on BKCa channels and signal transduction mechanism in corpus cavernosal smooth muscle cells

In this study, we investigated the effect of dopamine receptor agonists on potassium channels' activity and their signal transduction pathway in corporal smooth muscle cells. We used cultured human corporal smooth muscle cells. The whole cell and cell-attached configuration of the patch-clamp technique were used for electrophysiological recordings, and enzyme immunoassay was used for measuring cyclic AMP (cAMP) and cyclic GMP levels. Extracellular application of 10 microM dopamine and apomorphine significantly increased whole-cell K(+) currents by 283.5+/-55.7% (at +60 mV; n=12, P<0.001), 292.4+/-58.8.0% (at +60 mV; n=9, P<0.005), respectively. We confirmed that the increase in whole-cell currents was mainly due to activation of the tetraethylammonium-sensitive large conductance Ca(2+)-activated K(+) channels (BK(Ca) channels). Enzyme immunoassay indicated that dopamine and apomorphine stimulates cAMP levels in corporal smooth muscle cells in a concentration-dependent fashion. The activation of BK(Ca) channels by dopamine receptor agonists in corporal smooth muscle cells might be one of the mechanisms in inducing penile erection.

Chronic daily tadalafil prevents the corporal fibrosis and veno‐occlusive dysfunction that occurs after cavernosal nerve resection

To determine whether a long-term single daily oral dose of a longer half-life phosphodiesterase-5 (PDE5) inhibitor, tadalafil, has a similar effect to that of the shorter half-life PDE5 inhibitors sildenafil and vardenafil, and can prevent the fibrosis and resultant corporal veno-occlusive dysfunction (CVOD) occurring after cavernosal nerve (CN) injury. Male rats (10 per group) had either a sham operation, unilateral CN resection (CNR) or bilateral CNR, and were left untreated or given retrolingually 5 mg/kg per day of tadalafil. After 45 days, CVOD was assessed via cavernosometry, and the underlying corporal tissue changes were examined by immunohistochemistry and histochemistry (followed by quantitative image analysis), Western blots, and ad hoc methods. Tadalafil treatment normalized the low response to papaverine and high drop rate in the intracavernosal pressure measured by cavernosometry after CNR compared with sham-operated rats. Tadalafil also normalized the increase in penile shaft collagen content, and the reduction in corporal smooth muscle cell (SMC) content, SMC/collagen, and replication index, and improved the lower collagen III/I ratio and the increase in apoptotic index, caused by CNR, compared with sham operation. There were no effects of tadalafil on increased transforming growth factor beta1, inducible nitric oxide synthase and xanthine oxidoreductase levels. A long-term single daily dose of tadalafil prevented CVOD and the underlying corporal fibrosis in the rat caused by CN damage, as effectively as the previously reported continuous treatment with vardenafil or sildenafil, through a cGMP-related mechanism that appears to be independent of inducible nitric oxide synthase induction.

Ageing‐related corpora veno‐occlusive dysfunction in the rat is ameliorated by pioglitazone

To determine whether ageing-related changes in the penile corpora cavernosa, namely corporal veno-occlusive dysfunction (CVOD), loss of smooth muscle cells (SMCs), and excessive collagen deposition, can be ameliorated by the peroxisome proliferator-activated receptor gamma (PPARgamma) agonist pioglitazone, in a rat model of ageing as we have shown in a rat model of type 2 diabetes. Male Fischer 344 rats (16-18 months old) were fed chow containing 0%, 0.001% or 0.02% pioglitazone for 2 or 4.5 months, using 5 month old rats as 'young' controls. Functional changes were determined by dynamic-infusion cavernosometry (DIC). Histological changes were assessed by histochemistry and immunohistochemistry followed by quantitative image analysis and/or quantitative Western blot. Reactive oxygen species were estimated in blood. Pioglitazone at both doses reduced the high DIC 'drop rate' present in the untreated aged groups to the level seen in the young rats. The papaverine response was increased to young control levels by short-term high-dose pioglitazone and the long-term low-dose treatment, but not by the short-term low-dose treatment. Pioglitazone at all doses and durations of treatment failed to reverse the decreased corporal SMC/collagen ratio and SMC content, oxidative stress, or the elevated contents of collagen, or transforming growth factor beta1, seen in the aged penis, but did reduce the collagen III/I ratio, and at a high dose increased apoptosis. Both treatments inhibited the Rho-kinase system, by increasing Src homology region 2-containing protein tyrosine phosphatase and reducing Vav. PPARgamma were detected in corporal SMCs. Pioglitazone ameliorated ageing-related CVOD, possibly by a PPARgamma-mediated inhibition of Rho-kinase and not by a protective effect on the corporal smooth muscle.

Gene transfer of the KATP channel restores age‐related erectile dysfunction in rats

To determine if gene transfer of the ATP-sensitive potassium (K(ATP)) channel can reverse age-related erectile dysfunction in the rat, as the K(ATP) channel is an important subtype of potassium channels regulating smooth muscle tone. In an in vitro study, gene were transferred using cDNA of the K(ATP) channel in cultured human corporal smooth muscle (CSM) cells and human embryonic kidney (HEK) cells. After gene transfer, the activities of transferred channels were assessed by the patch-clamp technique. In an in vivo study, 15 old rats were used for groups of gene therapy and nine young adult rats were used as normal controls. The old rats were divided into three groups, i.e. controls and two gene-transfer groups (Kir6.1 + SUR2B and Kir6.2 + SUR2B). The intracavernosal pressure (ICP) response to cavernosal nerve stimulation was assessed after intracorporal injection with naked cDNA of the K(ATP) channel. The transgene expression of the K(ATP) channel was examined by reverse transcription-polymerase chain reaction (RT-PCR) in rats transfected with cDNA of Kir 6.1 and 6.2. The transferred gene of the K(ATP) channel was functionally active and appropriate for gene transfer. The mean (sem) ratio of ICP to systemic blood pressure in the gene-transfer groups, at 79.4 (1)% and 76.5 (2.6)% (both eight rats) was significantly higher than that in age-matched control rats, at 59.4 (3.3)% (eight), and similar to that in the young control rats, at 77.1 (2.7)% (nine). The RT-PCR showed expression of Kir6.1 and 6.2 genes in the transfected groups. In vivo gene transfer of the K(ATP) channel can physiologically restore erectile function in aged rats, and might be applicable to the development of new forms of therapy for treating human erectile dysfunction.

Erratum: Ageing causes cytoplasmic retention of MaxiK channels in rat corporal smooth muscle cells

Correction to: International Journal of Impotence Research (2007) 19, 371–377. doi:10.1038/sj.ijir.3901541 Following the online publication of the above article, the authors noted an error in one of the author names. The complete and correct author names are above.

Ageing causes cytoplasmic retention of MaxiK channels in rat corporal smooth muscle cells

The MaxiK channel plays a critical role in the regulation of corporal smooth muscle tone and thereby erectile function. Given that ageing results in a decline in erectile function, we determined changes in the expression of MaxiK, which might impact erectile function. Quantitative-polymerase chain reaction demonstrated that although there is no significant change in transcription of the alpha- and beta-subunits that comprise the MaxiK channel, there are significant changes in the expression of transcripts encoding different splice variants. One transcript, SV1, is 13-fold increased in expression in the ageing rat corpora. SV1 has previously been reported to trap other isoforms of the MaxiK channel in the cytoplasm. Correlating with increased expression of SV1, we observed in older rats there is approximately a 13-fold decrease in MaxiK protein in the corpora cell membrane and a greater proportion is retained in the cytoplasm (approximately threefold). These experiments demonstrate that ageing of the corpora is accompanied by changes in alternative splicing and cellular localization of the MaxiK channel.

Long-Term Continuous Treatment with Sildenafil Ameliorates Aging-Related Erectile Dysfunction and the Underlying Corporal Fibrosis in the Rat1

Aging-related erectile dysfunction is characterized by a loss of smooth muscle cells (SMCs) and fibrosis in the corpora cavernosa, and functionally by corporal veno-occlusive dysfunction (CVOD). Phosphodiesterase 5 (PDE5A) inhibitors, in part via upregulating inducible nitric oxide synthase (NOS2A), have antifibrotic properties in penile tissues. We aimed to determine whether in the aged rat the chronic long-term treatment with sildenafil ameliorates corporal SMC loss and fibrosis, stimulates NOS2A induction, and corrects the associated CVOD. Aged male rats (20 mo old) received sildenafil in their drinking water (20 mg/kg per day) or plain water for 45 days, and untreated young rats (5 mo old) served as controls (n = 8 per group). CVOD was assessed by dynamic infusion cavernosometry (DIC). Collagen:SMC (Masson trichrome) and collagen III:I (picrosirius red) ratios, SMC content (alpha-smooth muscle actin [ACTA2]), cell proliferation (proliferating nuclear antigen [PCNA]), apoptotic death (TUNEL), and NOS2A induction were measured by histochemistry and immunohistochemistry followed by quantitative image analysis. Collagen content was determined by hydroxyproline assay, and transforming growth factor beta-1 (TGFB1); xanthine oxidoreductase (XDH); ACTA2; NOS2A; and the Rho kinase inhibitor protein tyrosine phosphatase, nonreceptor type 11 (PTPN11), and activator, VAV, were measured by quantitative Western blot. In the aged rats treated with sildenafil, the erectile response by DIC was normalized, and the corporal SMC:collagen ratio and SMC number were increased. In addition, sildenafil reduced the corporal collagen content without affecting the collagen III:I ratio, increased the PCNA:apoptosis ratio, and stimulated NOS2A induction, although there was no effect on XDH, TGFB1, PTPN11, or VAV levels. These data show that long-term PDE5A treatment corrected CVOD in the aged rat and partially reversed the aging-related fibrosis and loss of SMC in the corpora cavernosa without affecting TGFB1 or PTPN11 levels, which are markers of oxidative stress. It may be speculated that similar effects may be achieved with this paradigm in men.

Potency after Radical Prostatectomy: From New Techniques to Better Results

Introduction and objectives Radical prostatectomy remains a commonly used procedure in the treatment of clinically localized prostate cancer. We critically analyzed current and future evidence-based strategies for preventing and managing postoperative erectile dysfunction (ED). Methods We conducted a systematic literature review using MEDLINE and CancerLit for the period from January 1997 through August 2005. We also assessed abstracts published in European Urology, the Journal of Urology, the International Journal of Impotence Research, and the Journal of Sexual Medicine, as well as official proceedings of internationally recognized scientific societies, during the same period. Results Patient selection and surgical technique are the major determinants of postoperative erectile function. Novel surgical approaches have been associated with increasing success in terms of restoring of erectile and urinary function after surgery. Pathophysiology of ED after radical prostatectomy includes chronic cavernosal hypoxia and apoptosis of corporeal smooth muscle cells, which might play a role in the development of cavernous arteriogenic and veno-occlusive dysfunction following radical prostatectomy. Recent evidence suggests that both pharmacological prophylaxis and treatment of postoperative ED are effective and safe. Conclusion In the hands of experienced high-volume surgeons, properly selected patients undergoing nerve-sparing radical prostatectomy should be able to achieve unassisted or medically assisted full erections in the postoperative period.

Lysophosphatidylcholine, a component of atherogenic lipoproteins, induces the change of calcium mobilization via TRPC ion channels in cultured human corporal smooth muscle cells

Hypercholesterolemia is a major risk factor for erectile dysfunction. To understand the mechanism(s) of hypercholesterolemia-induced erectile dysfunction, we studied the effect of lysophosphatidylcholine (LPC) on the membrane conductance of corporal smooth muscle cells. We used cultured human corporal smooth muscle cells. The intracelluar Ca2+ concentration ([Ca2+]i) and the influx of divalent cation was monitored by the ratio of fura-2 fluorescence (F(340/380)) and by the Mn2+-induced quenching rate of fura-2, respectively. The LPC-induced membrane current was characterized by the whole-cell patch-clamp technique and the molecular identity of suspected channels was probed by RT-PCR. LPC (20 microM) induced a statistically significant increase in F(340/380) to 119.9+/-3.9% of initial control (n=6) in corporal smooth muscle cells. The addition of 20 microM LPC accelerated the quenching rate of F360 by 59.5+/-11.8% (n=5). LPC activated nonselective cationic current (ILPC), similar to the known effects of phenylephrine in corporal myocytes. The size of ILPC at -60 mV was -55.3+/-6.3 pA (n=8). The transcript of transient receptor potential channel 6 (TRPC6) was detected in human corporal myocytes. We also found one splicing variant of TRPC6, TRPC6alpha. In conclusion, the present study suggests that the LPC, a major component of oxidized low-density lipoprotiens, increases calcium in corporal smooth muscle cells probably through activation of a TRPC6 channel and the increased [Ca2+]i by LPC via TRP channels is one of mechanisms for hypercholesterolemia-induced erectile dysfunction.

512The effect of testosterone on potassium channels in human corporal smooth muscle cells

512The effect of testosterone on potassium channels in human corporal smooth muscle cells

Intracorporal injection of hSlo cDNA restores erectile capacity in STZ-diabetic F-344 rats in vivo.

The ability of gene transfer with the pore-forming subunit of the human maxi-K channel (hSlo) to ameliorate the decline in erectile capacity commensurate with 12-24 wk of streptozotocin (STZ)-diabetes was examined in 181 Fischer-344 rats. A 2-mo period of STZ-diabetes was induced before gene transfer, and erectile capacity was evaluated by measuring the intracavernous pressure response (ICP) to cavernous nerve (CN) stimulation (ranging from 0.5 to 10 mA). In the first series of experiments, ANOVA revealed increased CN-stimulated ICP responses at 1 and 2 mo postinjection of 100 microg pcDNA-hSlo compared with control values. A second series of experiments further examined the dose dependence and duration of gene transfer. The ICP response to submaximal (0.5 mA) and maximal (10 mA) nerve stimulation was evaluated 3 or 4 mo postinjection of a single dose of pcDNA-hSlo ranging from 10 to 1,000 microg. ANOVA again revealed that hSlo overexpression was associated with increased CN-stimulated ICP responses compared with responses in corresponding control animals. Histological studies revealed no immune response to the presence of hSlo. PCR analysis documented that expression of both plasmid and transcript were largely confined to the corporal tissue. In the third series of pharmacological experiments, hSlo gene transfer in vivo was associated with iberiotoxin-sensitive relaxation responses to sodium nitroprusside in corporal tissue strips in vitro. The latter data indicate that gene transfer produces functional maxi-K channels that participate in the modulation of corporal smooth muscle cell tone. Taken together, these observations suggest a fundamental diabetes-related change in corporal myocyte maxi-K channel regulation, expression, or function that may be corrected by expression of recombinant hSlo.

Physiology and biochemistry of erections.

The physiology of erection has received intense clinical and basic research scrutiny over the past two decades. This massive effort has led to a much clearer understanding of the macroscopic aspects of erection as well as identification of the prominent features of the etiology of erectile dysfunction (ED). However, it is clear that the devil is in the details of the erectile process. Therefore, to truly understand the precise mechanistic basis for erection and ED, much more still must be learned about how the biochemical cascades in the corporal smooth muscle cells are integrated to produce a normal erection, or how they are altered to result in ED. The ultimate goal of these basic research and clinical efforts will be to provide a rational scientific basis for mechanism-based, patient-specific therapies for ED. This article reviews fundamental aspects of the physiology of erection and summarizes the most recent information available concerning the putative biochemical correlates of these physiologic events.