Cavernosum Smooth Muscle Research Articles

Efficient use of penile Doppler ultrasound for investigating men with erectile dysfunction.

Vasculogenic erectile dysfunction is the most common type of erectile dysfunction, and penile Doppler ultrasound (PDUS) is a useful tool to assess erectile hemodynamics in the clinician's effort to discuss prognosis and management strategies with the patient. We herein describe the PDUS protocol used at our center, including indications, technique, and data interpretation. We describe our institutional experience with PDUS and discuss it in the context of a contemporary review of the literature for this investigation. Our institutional PDUS protocol. To perform PDUS properly, adequate training, equipment, setting, technique, and interpretation are critical. The accuracy of PDUS is entirely predicated on achieving complete cavernosal smooth muscle relaxation. A redosing protocol optimizes the reliability and reproducibility of the hemodynamic data acquired during PDUS. A rigidity-based assessment is performed, and patients are scanned according to the erection rigidity achieved (full hardness) or by administration of maximum dose of the vasoactive agent. Peak systolic velocity is considered a measure of arterial inflow (normal, >30cm/s), while end diastolic velocity evaluates the veno-occlusive mechanism (normal, <5cm/s). After the procedure, the patient is evaluated to confirm detumescence. If the patient has a persistent penetration rigidity erection, intracavernosal phenylephrine is administered; however, if detumescence is not achieved with intracavernosal phenylephrine injections alone, corporal aspiration is potentially performed. PDUS is a valuable minimally invasive tool for erectile hemodynamics assessment and an accurate assessment of such, provided that complete cavernosal smooth muscle relaxation is achieved.

Low Serum Testosterone as a Poor Prognostic Marker in Urethral Stricture: A Single-Center Prospective Longitudinal Study.

Introduction Androgens play a key role in modulating periurethral and preputial vascularity, cavernosal smooth muscle integrity, and penile growth. As a result, low testosterone may adversely affect the severity and outcome of urethral stricture patients. So, to find out the hormonal influence on the clinical outcome of urethral stricture we conducted a prospective longitudinal study at our institute. Methods The study was conducted at the Department of Urology, Institute of Post Graduate Medical Education & Research (IPGMER), Kolkata, India, from February 2023 to September 2023. This study was approved by the Institutional Ethics Committee atIPGMER, Kolkata with the approval number IPGMER/IEC/2023/436. Hormonal levels in patients with diagnosed non-traumatic urethral stricture were compared with patients without stricture. Patients with any overt hormonal abnormality or androgen-secreting tumor were excluded. A morning 10 cc blood sample was collected for testosterone, follicle-stimulating hormone, luteinizing hormone, and thyroid-stimulating hormone. The association of hormonal levels was measured in both groups and compared statistically. Any association of hypogonadism (testosterone <300 ng/dL) with respect to length, severity, and recurrence of urethral stricture was also studied. Results Forty patients with urethral stricture and same number of patients without stricture were included in the study. The mean testosterone level was found to be significantly low in patients with stricture (386 ng/dL vs 660 ng/dL). The age-wise distribution also showed low mean testosterone compared to patients without stricture. The incidence of hypogonadism is also found to be higher in stricture patients (47.5% vs 27.5%). It was also observed low testosterone is more prevalent in pan-anterior stricture (10/40) and long-segment stricture(>2 cm). Patients with stricture were also followed up for 6 months for recurrence of symptoms. Thirteen patients had recurrence. Patients with recurrence had significantly low serum testosterone (272 ng/dL vs 440 ng/dL). Conclusion Our study documented stricture patients with low serum testosterone have poor outcomes. Low testosterone level is strongly associated with longer stricture and increased risk of stricture recurrence.

(061) Analysis of BMP4 and GREM1 as Targets of SHH Signaling and Downstream Regulators of the Collagen Axis in the Penis

Abstract Introduction Cavernous nerve (CN) injury caused by prostatectomy and diabetes, initiates a remodeling process (smooth muscle apoptosis and increased collagen) in the corpora cavernosa of the penis of patients and animal models that is an underlying cause of erectile dysfunction (ED). The Sonic hedgehog (SHH) pathway plays an essential role in the response of the penis to denervation, as collagen increases with SHH inhibition and decreases with SHH treatment. Part of the mechanism of how SHH impacts collagen may involve bone morphogenetic protein four (BMP4) and GREMLIN (GREM1). Microarray analysis identified increased GREM1 (BMP4 antagonist) in corpora cavernosa of ED patients and SHH is a regulator of GREM1 in the limb bud. Objective We will examine the relationship between SHH, BMP4, GREM1 and collagen in penis of ED patients and rat models of CN injury, SHH inhibition, and SHH, BMP4 and GREM1 treatment. Methods Corpora cavernosa of Peyronie’s (control), prostatectomy and diabetic ED patients was obtained (n=26). Adult Sprague Dawley rats (n=90) underwent either: 1. CN crush (1-7 days) or sham control, 2. CN injury and BMP4, GREM1 or MSA (control) protein treatment via Affi-Gel beads or peptide amphiphile (PA) for 14 days, 3. 5E1 SHH inhibitor, IgG and PBS (controls), or SHH protein treatment for 2-4 days. Immunohistochemical and western analysis for BMP-4 and GREM1, and collagen analysis by hydroxyproline and trichrome stain were performed. Results BMP4 and GREM1 proteins were identified in corpora cavernosal smooth muscle of prostatectomy, diabetic and Peyronie’s patients, and in rat penis, sympathetic nerve fibers, perineurium, blood vessels, and urethra. Collagen decreased 25.4% in rats with CN injury and BMP4 PA treatment (p=0.02) and increased 61.3% with CN injury and GREM1 treatment (p=0.005). Trichrome showed increased collagen in rats treated with GREM1. BMP4 increased smooth muscle. Western identified increased BMP4 and GREM1 in corpora cavernosa of prostatectomy and diabetic patients, and after CN injury (1-2 days) in our rat model. Localization of BMP4 and GREM1 changed with SHH inhibition. SHH treatment increased BMP4 and GREM1. Aging did not impact BMP4 and GREM1 localization. Conclusions BMP4 and GREM1 are downstream targets of SHH that impact collagen and may be useful in collaboration with SHH to prevent penile remodeling and ED. Better understanding of penile remodeling and how fibrosis occurs is essential for development of novel therapies for ED. Disclosure No.

(066) Targets of SHH Signaling, BMP4 and GREM1, are Downstream Regulators of Smooth Muscle in the Penis

Abstract Introduction A key aspect of erectile dysfunction (ED) development is loss of innervation to the penis. The cavernous nerve (CN) is frequently damaged in prostatectomy and diabetic patients with ED, initiating changes in penile morphology including an acute and intense phase of apoptosis in penile smooth muscle followed by increasing collagen and fibrosis. This remodeling process alters penile architecture so that there is less smooth muscle in the corpora cavernosa, and what remains is less compliant/able to relax in response to normal neurotransmitter signaling, and ED results. Thus, novel therapies are needed which target the underlying remodeling process. We have shown previously that Sonic hedgehog (SHH) is a critical regulator of penile smooth muscle, and that SHH protein treatment suppresses penile remodeling after CN injury through an unknown mechanism. Objective We examine if part of the mechanism of how SHH preserves smooth muscle after CN injury involves bone morphogenetic protein 4 (BMP4) and GREMLIN (GREM1). Methods Primary cultures of smooth muscle cells were established from patients who had prostatectomy, diabetes, hypertension and Peyronie’s (control, n=18). Cultures were characterized by immunohistochemical analysis (IHC) for ACTA2, CD31, P4HB and nNOS. Smooth muscle cell growth was quantified in response to BMP4 and GREM1. Adult Sprague Dawley rats that were uninjured or underwent CN crush injury, were treated with BMP4, GREM1 or MSA (control) proteins via Affi-Gel beads (n=16) or peptide amphiphile (PA, n=26) for 3 and 14 days, and trichrome stain was performed. Sprague Dawley rats underwent CN injury (n=9) and SHH PA treatment for 1, 2 and 4 days (n=9) and western was performed assaying for BMP4 and GREM1 proteins in comparison to sham (n=3) controls. Adult Sprague Dawley rats were treated with 5E1 SHH inhibitor (n=6) or IgG (control, n=6) for 2 and 4 days, and BMP4 and GREM1 localization were examined by IHC. Statistics were performed by ANOVA with Scheffe’s posthoc test. Results Patient primary smooth muscle cultures were established. BMP-4 increased patient smooth muscle cell growth and GREM1 decreased growth. In rats BMP4 treatment via Aff-Gel beads and PA increased smooth muscle growth at 3 and 14 days of treatment. GREM1 treatment caused collagen and smooth muscle growth at 3 days, which switched to primarily collagen at 14 days. CN injury increased BMP4 and GREM1. SHH PA altered western band size suggesting alternative cleavage and range of signaling. SHH inhibition in rats changed BMP4 and GREM1 localization. Conclusions BMP4 treatment increases smooth muscle in patient cultures and in our rat model. BMP4 and GREM1 mediate a switch from smooth muscle to collagen induction. SHH treatment alters BMP4 and GREM1 localization and range of signaling, which can affect penile morphology. Whether GREM1 directly effects collagen induction or acts strictly through inhibition of BMP4 is unknown and requires further study. Part of the mechanism of how SHH regulates corpora cavernosal smooth muscle, involves BMP4 and GREM1. Understanding how SHH PA impacts penile morphology after CN injury will aid development of ED therapies. Disclosure No.

A possible mechanism of erectile dysfunction in coronavirus disease-19: Cavernosal smooth muscle damage: A pilot study

IntroductionStudies have reported that coronavirus disease 2019 (COVID-19) may cause erectile dysfunction (ED), however, its role in the pathophysiology of ED has not yet been fully elucidated. We aimed to elucidate COVID-19's effects on cavernosal smooth muscle, which has a pretty important role in erection physiology, by corpus cavernosum electromyography (cc-EMG). Materials and methodsTwenty-nine male patients aged 20–50 years who applied to the urology outpatient clinic due to ED were included in the study. Nine patients that had COVID-19 and were treated as outpatients were classified as group 1, 10 patients who were hospitalized due to COVID-19 were classified as group 2, and 10 patients who did not have COVID-19 were classified as the control group (group 3). Patients underwent diagnostic evaluation including International Index of Erectile Function (IIEF)-5 form, penile color Doppler ultrasonography (CDUS), cc-EMG, and fasting serum levels of reproductive hormones (07–11am). ResultsAccording to penile CDUS and hormonal values results, there was no significant difference between the groups. According to cc-EMG results, amplitudes and relaxation capacities of the cavernosal smooth muscle of patients in group 3 were significantly higher than those in the other groups. ConclusionsCOVID-19 can cause ED not only by psychogenic and hormonal factors but also with cavernosal smooth muscle damage.Clinical Trial Registration Number: NCT04980508.

(202) Exploring Erectile Hemodynamics in Men Complaining of Hard-Flaccid Syndrome (HFS)

Abstract Introduction There is increasing awareness of a condition, vernacularly termed HFS, in which men complain of subjective and palpable abnormalities in the flaccid penis (hardness, heaviness) often associated with erectile dysfunction (ED). Objective We attempted to define erectile hemodynamic profiles in this population. Methods The study population consisted of (i) males ≥18 years of age (ii) who presented with self-diagnosed HFS (iii) who underwent comprehensive history and examination (including genital and lower extremity neurological) and (iv) penile duplex Doppler ultrasound (PDDU). PDDU was performed using redosing of vasodilators. PSV ≥35 cm/s and EDV ≤3 were considered normal. Patients with Peyronie’s disease or a history of intracavernosal injection therapy were excluded. Results 88 men were analyzed. Mean age was 28 ± 12 years. Median number of vascular comorbidities was zero (0, 1). 66 (75%) had a history of depression or anxiety, 60 of whom had used pharmacotherapy for such at some time and 40 were actively using anxiolytics or anti-depressants at time of evaluation. No patient had a history of neurological conditions and neurological exam was normal in all. Mean duration of symptoms was 14 ± 20 months. Median number of prior physician consultations was 1 (1,4). All complained of impaired erectile function, 50% being incapable of having penetrative sexual relations. 91% had or were using PDE5 inhibitors. On examination all patients had a hypertonic flaccid penis with excellent stretch on traction. At PDDU, median number of penile injections was 2 (1, 3). 50% of men required phenylephrine reversal. All patients had normal PSV with 2/88 having abnormal EDV values, both of whom required reversal. Mean PSV was 45 ± 35 cm/s, mean EDV was 2 ± 11 cm/s. No patient had elevated echogenicity on B-mode scanning. Conclusions In this analysis, all men presenting with self-reported HFS had normal hemodynamics and an absence of fibrosis, suggesting the absence of vascular or gross cavernosal smooth muscle structural changes. Disclosure No

(343) Predictors of Corporo-venocclusive Dysfunction (CVOD) in Men with Bilateral Nerve-sparing Radical Prostatectomy (RP)

Abstract Introduction Nerve-sparing status (NSS) during RP is a significant predictor of erectile function recovery (EFR). Poor nerve sparing is associated with collagenization of cavernosal smooth muscle and CVOD development with typically poor response to PDE5 inhibitors (PDE5i) and intracavernosal injection therapy (ICI). Objective This study aimed to define factors predictive of CVOD in men with bilateral NS RP. Methods Study population consisted of (i) men who underwent bilateral NS (defined by the surgeon using an institutional grading system) RP (ii) with poor response to PDE5i and ICI (iii) ≥2 years post-RP (iv) who underwent penile duplex Doppler ultrasound (DUS) (v) using a vasoactive agent redosing schedule. CVOD diagnosis was made with end-diastolic velocity (EDV) values ≥5 cm/sec bilaterally. ADT and radiation therapy were exclusions. We used multivariable modeling to define factors predictive of CVOD. Factors included in the model: patient age, history of diabetes, history of obstructive sleep apnea, number of vascular comorbidities, baseline EF, peri-RP total testosterone (T) level, and total T at DUS (within 3 months). Results 201 men with a mean age of 66 ± 7 years. 25% of the patients reported 3 or more vascular comorbidities, 17% had diabetes, 36% obstructive sleep apnea, 39% were current or former smokers. Mean baseline PSA and pre-RP total T was 6.7 ± 6 ng/mL and 410 ± 211 ng/dL, respectively. Median baseline EFD score was 24 (13, 29). 49% of the men had an erection less than penetration rigidity during the DUS procedure with a mean total dose administered of 87 ± 21 units of trimix. 69% received 100 units. CVOD was diagnosed in 76%. Mean total T at DUS was 386 ± 177 ng/dL. 32% had low T. Significant predictors of CVOD after RP in men with BL NSS were: OSA (OR = 3.8, 95% CI = 1.7-8.7), ≥ 3 vascular comorbidities (OR = 5.0, 95% CI = 1.7-14.8), age at the time of DUS (per 10 years increase, OR = 1.9, 95% CI = 1.2-3.0), and pre-RP EF domain score (per 1 unit increase, OR = 0.93, 95% CI 0.88-0.98). There was a signal that in a larger sample size, diabetes may be a significant predictor of CVOD in this group of patients (OR = 2.7, 95% 0.9-8.0). Conclusions Baseline clinical comorbidities pre-RP have a significant impact on EFR post-RP, in particular, the presence of venous leak. Despite BL NSS, older patients, men with OSA, higher number of comorbidities pre-RP, and lower pre-RP EF increased the likelihood of developing CVOD post-RP. Disclosure No

(091) RNAseq and Pathway Analysis of Rat Penis with Cavernous Nerve Injury and Sonic Hedgehog Treatment by Peptide Amphiphile

Abstract Introduction Erectile dysfunction (ED) treatments are minimally effective in prostatectomy and diabetic patients due to injury to the cavernous nerve (CN). When the CN is injured, loss of innervation initiates penile remodeling, which includes abundant apoptosis in corpora cavernosal smooth muscle, followed by fibrosis. Decreased smooth muscle makes the erectile tissue less able to respond to normal neurotransmitter signaling, and ED results. When developing novel therapies, the goal is to prevent the apoptotic response and penile remodeling, to preserve erectile function. Our previous studies identify Sonic hedgehog (SHH) as a key mediator of penile remodeling. We examine what signaling mechanisms are altered with CN injury, and how SHH impacts changes in signaling to prevent penile remodeling and ED. Objective We performed RNA seq analysis and subsequent pathway analysis, to identify changes in gene expression and signaling pathways that are altered in response to cavernous nerve injury and with SHH treatment by peptide amphiphile (PA), in a rat prostatectomy model of ED. Methods Corpora cavernosa were obtained from Sprague Dawley rats (n=36) that underwent bilateral CN crush, CN crush plus SHH treatment of the penis by PA, or sham surgery (controls). Rats were sacrificed 1, 2 and 4 days after surgery. RNA was extracted using TRIzol, DNase treated, and purified by Qiagen mini kit. RNAseq analysis was performed, and subsequent directional pathway enrichment analysis was examined using Ingenuity software. A two-fold change with a q-value &amp;lt;0.05 was used as a threshold for differential expression. Z value measures were used to predict altered activity for the identified enrichments. Results mRNA representing 523 genes were significantly differentially expressed (&amp;gt;2 fold; Q&amp;lt;0.05) in CN crushed corpora cavernosal tissue relative to sham controls. Ingenuity analysis showed altered activity in canonical signaling and disease-function enrichments. These changes include increased necrosis and netrin, and decreased Integrin linked kinase, integrin, actin cytoskeleton, acute phase response signaling, cell movement, angiogenesis, and myeloid cell movement activity. With SHH PA treatment at the time of CN injury, these changes were reversed or significantly diminished. In addition, SHH PA impacted known mediators of ED by reducing apoptosis, and increasing cell survival, cell viability, muscle cell proliferation, VEGF, p38 MAPK, and PI3K/AKT pathways. Conclusions RNAseq and pathway analysis identified changes in gene expression and pathway signaling that is associated with CN injury in a rat prostatectomy model. Pathway analysis showed increases in processes associated with cell death and a loss of functions necessary for maintenance of corpora cavernosal architecture. Most of the CN injury associated changes were reversed or greatly diminished with SHH treatment of the penis at the time of CN injury by PA. These results support the significant potential of SHH PA to be developed as a preventative treatment for ED in prostatectomy patients. The miriad of altered signaling pathways that were identified with CN injury offer multiple, novel avenues for intervention independently and in combination with SHH PA treatment. Disclosure No

(084) Analysis of BMP4 and GREMLIN as targets of SHH signaling and regulators of the collagen axis in the penis.

Abstract Introduction Increased collagen deposition occurs in erectile dysfunction (ED) patients and animal models, and the underlying causes of these fibrotic changes have not been well defined. The Sonic hedgehog (SHH) pathway is critical for the response of the penis to denervation. Decreased SHH in the penis due to cavernous nerve (CN) injury, causes smooth muscle apoptosis and ED. As smooth muscle is lost, increased collagen occurs by a largely unknown mechanism. We have shown that collagen abundance is responsive to SHH signaling in the penis. Microarray analysis identified increased GREMLIN (BMP4 antagonist) in corpora cavernosa of ED patients, and SHH is a regulator of GREMLIN in the limb bud. Objective We hypothesize that SHH regulation of collagen occurs through a BMP4/GREMLIN dependent mechanism and will examine BMP4 and GREMLIN signalling in corpora cavernosa of ED patients and a rat prostatectomy model. Study of how collagen induction occurs after prostatectomy would be useful to determine the underlying mechanism of penile remodeling. Methods Corpora cavernosal tissue (n=36) from prostatectomy, diabetic and Peyronie’s (control) patients were examined by immunohistochemical and western analysis for BMP-4 and GREMLIN proteins. Localization was determined by dual staining with ACTA2 and CD31. Collagen was quantified by hydroxyproline assay and trichrome stain with Image J analysis. Primary cultures of patient corpora cavernosa were established and treated with BMP4 and GREMLIN proteins, and smooth muscle cell growth was quantified. Sprague dawley rat penis was treated in vivo with SHH protein by peptide amphiphile, 5E1 SHH inhibitor, BMP4 and GREMLIN via Affi-Gel beads. Collagen was quantified. Results BMP-4 and GREMLIN were identified in smooth muscle of Peyronie’s, prostatectomy and diabetic ED patients. Western analysis identified increased BMP4 and GREMLIN proteins in prostatectomy and diabetic patients where collagen is increased. Primary culture of corpora cavernosal smooth muscle cells showed an increase in growth with BMP4 and decreased growth with GREMLIN treatment. This trend was more pronounced in cells from hypertension and cardiovascular disease patients. BMP4 and GREMLIN proteins increased 1-2 days after CN injury in a rat prostatectomy model that showed decreasing SHH in the penis. SHH inhibition in the penis increases BMP4, GREMLIN and collagen. SHH treatment of the penis after CN injury in the rat decreases collagen, BMP4 and GREMLIN. Conclusions SHH is decreased in corpora cavernosa of prostatectomy and diabetic ED patients while BMP4 and GREMLIN are increased. Decreased SHH increases BMP4 protein and collagen, suggesting a possible mechanism for fibrotic penile remodelling after CN injury. This study is the first step in understanding the mechanism of how SHH regulates collagen abundance in the penis, which is important for clinical translation. Disclosure No

Ethnological validation of Ashwagandha (Withania somnifera L. Dunal) ghrita as ‘Vajikarana Rasayana’: In-silico, in-vitro and in-vivoapproach

Ethnopharmacological relevanceProcessing cow ghee (clarified butterfat) with therapeutic herbs, i.e. ghrita, is recognized for augmenting the therapeutic efficacy of plant materials. Ashwagandha ghrita (AG) is an effective Ayurvedic formulation consisting of Indian ginseng, i.e., Withania somnifera (L.) Dunal, the main constituent used to treat infertility, weakness, gynaecological disorders, and general debility. ObjectivesThe present investigation was undertaken to corroborate the ethnopharmacological claim of AG as ‘Vajikarana Rasayana’ for its aphrodisiac potential using bioinformatics (in-silico) and experimental (in-vitro and in-vivo) approaches. MethodsAG was formulated as per the methods reported in Ayurved sarsangraha. AG was further subjected to HPLC, GCMS analysis, and biological (acute toxicity and aphrodisiac) assessment per the standard procedures. Thirty-eight bioactives of Indian ginseng were subjected to computational studies (molecular docking and network pharmacology) to confirm the plausible mechanism. ResultsAG was found to be safe up to 2000 mg/kg body wt., and it showed dose-dependent upsurge (p < 0.01 and p < 0.05, wherever necessary) in mount and intromission frequency, genital grooming, and anogenital sniffing at 150 and 300 mg/kg body weight suggesting aphrodisiac activity. In-vitro studies demonstrated significant relaxation of the Corpus Cavernosal Smooth Muscle at all concentrations in a dose-dependent manner. Furthermore, the results of molecular modelling studies were in agreement with the biological activity and showed interaction with phosphodiesterase-5 as a possible target. ConclusionAG exhibited an aphrodisiac effect and substantiated the traditional claim of Indian ginseng-based ghrita formulation as ‘Vajikarana Rasayana’.

Treatment with Resveratrol Reverses Priapism in the Sickle Cell Mouse

Introduction: Sickle cell disease (SCD) men display priapism. Townes and Berkeley SCD mice models are widely studied in basic sciences because they express 100% HbS in their circulating erythrocytes and exhibit features of priapism, and, as in SCD humans, this appears to be due to decreased nitric oxide (NO) signaling and increased oxidative stress, which leads to lower PDE5 in the penis. Thus, when an erectile stimulus occurs in vivo, cGMP accumulates into the cavernosal smooth muscle cells, rendering the penile vasculature uncontrollably dilated, and penile erection persists (i.e., priapism) as cGMP is not degraded as a consequence of PDE5 downregulation. Moreover, oxidative/nitrosative stress has been associated with priapism in SCD mice.Resveratrol (trans-3,5,4′-trihydroxystilbene) is a natural phytoalexin product with potent antioxidant activity found in peanuts, berries, and grapes. In this study, since decreased NO bioavailability and increased oxidative stress leads to lower PDE5 in the penis, we aimed to evaluate the effects of resveratrol on functional and molecular alterations of erectile function in SCD mice. We have focused on the dysregulated NO-cGMP-PDE5 pathway and oxidative/nitrosative stress markers (4-HNE and 3-nitrotyrosine) in the erectile tissue of SCD. Methods: Wild type (WT, C57BL/6) and Townes SCD transgenic male mice (3-5 mo old) were treated with resveratrol (100 mg /kg/day) or its vehicle daily for two weeks via gavage. Corpus cavernosum (CC) strips were mounted in isolated organ baths, and the relaxing responses to acetylcholine (ACh; endothelium-dependent response) and sodium nitroprusside (SNP; endothelium-independent response), as well as electrical-field stimulation (EFS; nitrergic relaxation), were obtained in CC strips precontracted with the α1-adrenergic receptor agonist phenylephrine (3-10 µM). Results: The cumulative addition of ACh (0.001-10 µM) produced concentration-dependent CC relaxations in WT and SCD groups, but the ACh potency (pEC50) value was higher (P<0.05) in CC of SCD compared to WT mice, which was reversed by resveratrol. Likewise, the nitrergic relaxation induced by EFS was also higher (P<0.05) in SCD mice compared to control mice (4 Hz: 30 ± 8 and 80 ± 5 %, respectively; n=5), which was reversed by resveratrol treatment. Similarly, SNP (0.01- 10 µM) produced concentration-dependent CC relaxations in WT and SCD groups, but, again, the maximal relaxations elicited by this agent were higher (P<0.05) in SCD (90 ± 1%) group compared to the WT mice (65 ± 6%), which were fully restored to WT values with resveratrol treatment (n=6). The mRNA expression for the PDE5 was reduced (P<0.05) by approximately 57% in penile tissues from SCD compared to the WT group, respectively. Resveratrol treatment restored (P<0.05) the mRNA expression of PDE5 in the penises from the SCD group (n=5-6). The protein expressions for 4-HNE and 3-nitrotyrosine (3-NT) were significantly higher (P<0.05) in erectile tissues from SCD compared to the WT group, which were reduced (P<0.05) by resveratrol treatment (n=6). In WT mice, PDE5, 4-HNE, and 3-NT protein expressions were not affected by resveratrol, as well as CC relaxations induced by ACh, SNP, and EFS. Conclusion: Resveratrol treatment reduced enhanced NO-mediated CC relaxations along with normalization of PDE5 expression and reduced oxidative stress markers expressions (3-NT and 4-HNE) in the penises from SCD. Therefore, resveratrol may constitute an additional strategy to prevent priapism in SCD. Financial Support: FAPESP.

Recent advances in understanding and treating priapism.

Priapism is a rare condition that can lead to long-term erectile dysfunction if left untreated. It is one of the few urological emergencies that require prompt medical intervention. Priapism refers to a penile erection that lasts for more than 4 hours and is unrelated to sexual stimulation or orgasm. The aims of immediate intervention for ischaemic priapism are to resolve the painful erection and preserve the cavernosal smooth muscle function. The aim of this review is to evaluate the latest advances in the management of priapism. Despite the continuous challenge in providing an optimal treatment for this rare urological condition, our understanding and management of it have been advanced by decades of clinical and basic science research. Proximal shunts (Quackels or Grayhack) are no longer routinely performed. Distal shunt procedures are currently the most commonly used techniques. A novel penoscrotal decompression technique has recently been described. Ischaemic priapism can be managed conservatively in most cases with the preservation of erectile function. In cases where ischaemic priapism has persisted for more than 36 hours, the majority will develop erectile dysfunction. Early penile prosthesis with thorough patient counselling should be considered in such cases. In some cases of long-standing non-ischaemic priapism, patients can develop fibrosis within the distal corpora, and, therefore, early treatment with super-selective embolisation is required to prevent this.

Sonic Hedgehog Signaling in Primary Culture of Human Corpora Cavernosal Tissue From Prostatectomy, Diabetic, and Peyronie's Patients

BackgroundCavernous nerve (CN) injury causes penile remodeling, including smooth muscle apoptosis and increased collagen, which results in erectile dysfunction (ED), and prevention of this remodeling is critical for novel ED therapy development. AimWe developed 2 peptide amphiphile (PA) hydrogel delivery vehicles for Sonic hedgehog (SHH) protein to the penis and CN, which effectively suppress penile distrophic remodeling (apoptosis and fibrosis), in vivo in a rat CN injury model, and the aim of this study is to determine if SHH PA can be used to regenerate human corpora cavernosal smooth muscle deriving from multiple ED origins. MethodsCorpora cavernosal tissue was obtained from prostatectomy, diabetic, hypertension, cardiovascular disease and Peyronie's (control) patients (n = 21). Primary cultures (n = 21) were established, and corpora cavernosal cells were treated with SHH protein, MSA (control), 5E1 SHH inhibitor, and PBS (control). Growth was quantified by counting the number of cells at 3–4 days. Statistics were performed by ANOVA with Scheffe's post hoc test. Concentration of SHH protein for maximal growth was optimized, and a more active SHH protein examined. OutcomesCultures were characterized by immunohistochemical analysis with ACTA2, CD31, nNOS and P4HB, and smooth muscle was quantified in comparison to DAPI. ResultsCultures established were >97% smooth muscle. SHH protein increased growth of smooth muscle cells from prostatectomy, diabetic, and Peyronie's patients in a similar manner (49%–51%), and SHH inhibition decreased growth (20%–33%). There was no difference in growth using 25 ug and 10 ug SHH protein, suggesting a threshold concentration of SHH protein above which smooth muscle growth is enhanced. A more active lipid modified SHH peptide further enhanced growth (15%), indicating a more robust growth response. SHH increased growth in smooth muscle cells from hypertension (37%) and cardiovascular disease (32%) patients. SHH protein increased growth under normal and high glucose conditions, suggesting that high glucose conditions that may be present in under controlled diabetic patients would not detract from SHH regenerative capacity. Clinical ImplicationsSHH PA would be beneficial to enhance smooth muscle regeneration in patients with ED of multiple etiologies. Strengths and LimitationsUnderstanding how human corpora cavernosal tissue responds to SHH treatment is critical for clinical translation of SHH PA to ED patients. ConclusionCorpora cavernosal smooth muscle from all ED patients responded to SHH treatment with increased growth.Stupp, SI. Sonic Hedgehog Signaling in Primary Culture of Human Corpora Cavernosal Tissue From Prostatectomy, Diabetic, and Peyronie's Patients. J Sex Med 2022;19:1228–1242.

MP38-06 SONIC HEDGEHOG SIGNALING IN CORPORA CAVERNOSAL CELLS FROM PROSTATECTOMY, DIABETIC, HYPERTENSION AND PEYRONIE’S PATIENTS WITH ERECTILE DYSFUNCTION

You have accessJournal of UrologyCME1 May 2022MP38-06 SONIC HEDGEHOG SIGNALING IN CORPORA CAVERNOSAL CELLS FROM PROSTATECTOMY, DIABETIC, HYPERTENSION AND PEYRONIE’S PATIENTS WITH ERECTILE DYSFUNCTION Sarah Martin, Timothy Searl, Samuel Ohlander, Kevin McVary, and Carol Podlasek Sarah MartinSarah Martin More articles by this author , Timothy SearlTimothy Searl More articles by this author , Samuel OhlanderSamuel Ohlander More articles by this author , Kevin McVaryKevin McVary More articles by this author , and Carol PodlasekCarol Podlasek More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002592.06AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Erectile dysfunction (ED) treatments are minimally effective in prostatectomy and diabetic patients due to injury to the cavernous nerve. With denervation the critical smooth muscle undergoes apoptosis and the penis becomes fibrotic, thus altering corpora cavernosal architecture. In order to devise novel ED therapies, prevention of corpora cavernosal remodeling is critical. Sonic hedgehog (SHH) treatment by peptide amphiphile (PA) nanofiber hydrogel, suppresses smooth muscle apoptosis and improves erectile function in a rat CN injury ED model. We examine if human corpora cavernosal smooth muscle responds in a similar manner to SHH treatment.In this study we: 1.) Examine if human corpora cavernosal smooth muscle cells from primary culture of ED patient tissues, respond to Sonic hedgehog (SHH) treatment by increasing growth rate, as in our in vivo animal model. 2.) Determine if human corpora cavernosal cells from patients with differing underlying mechanisms of ED development (prostatectomy, diabetes, hypertension, cardiovascular disease), respond similarly to SHH treatment. 3.) Identify if organ culture conditions (glucose) affect growth of corpora cavernosal smooth muscle and the response to SHH signaling. 4.) Examine the vascular component of ED (hypertension and cardiovascular disease). METHODS: Human corpora cavernosal tissue was obtained from prostatectomy, diabetic, hypertension, cardiovascular disease and Peyronie’s (control) patients (n=40) that were under going prosthesis implant to treat ED. Primary cultures (n=17) were established, and corpora cavernosal cells from passage 3-4 were treated with SHH protein, MSA (control), 5E1 SHH inhibitor, and PBS (control). Growth was quantified by counting the number of cells using a hemocytometer at 3-4 days. The concentration of SHH protein for maximal growth was examined, in addition to a lipid modified more active SHH peptide. RESULTS: SHH treatment increased smooth muscle growth in human corpora cavernosal cells from prostatectomy, diabetic, and Peyronie’s patients in a similar manner (43-53%). SHH inhibition decreased smooth muscle growth (24-32%). There was no difference in growth using 25ug and 10ug SHH peptide. A more active (150X) SHH peptide further enhanced growth (20%). SHH protein increased growth more in diabetic smooth muscle under high glucose conditions. SHH inhibition was less effective under high glucose conditions. SHH treatment increased growth less in cells from hypertension and cardiovascular disease patients. CONCLUSIONS: Corpora cavernosa smooth muscle from prostatectomy, diabetic, and Peyronie’s patients increased in response to SHH treatment in a similar manner, suggesting that SHH treatment would be beneficial to enhance smooth muscle regeneration in patients with ED of multiple origins. There is a threshold concentration of SHH protein above which smooth muscle growth is enhanced. Lipid modified SHH engendered a more robust growth response. High glucose conditions that may be present in under controlled diabetic patients would not detract from SHH usefulness as a protectant for corpora cavernosal morphology. Understanding how human corpora cavernosal tissue responds to SHH treatment is critical for clinical translation to ED patients. Source of Funding: NIH/NIDDK DK101536 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e620 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Sarah Martin More articles by this author Timothy Searl More articles by this author Samuel Ohlander More articles by this author Kevin McVary More articles by this author Carol Podlasek More articles by this author Expand All Advertisement PDF DownloadLoading ...

Sonic hedgehog signaling in corpora cavernosal cells from prostatectomy, diabetic, hypertension and Peyronie's patients with erectile dysfunction

ABSTRACT Introduction Erectile dysfunction (ED) treatments are minimally effective in prostatectomy and diabetic patients due to injury to the cavernous nerve. With denervation the critical smooth muscle undergoes apoptosis and the penis becomes fibrotic, thus altering corpora cavernosal architecture. In order to devise novel ED therapies, prevention of corpora cavernosal remodeling is critical. Sonic hedgehog (SHH) treatment by peptide amphiphile (PA) nanofiber hydrogel, suppresses smooth muscle apoptosis and improves erectile function in a rat CN injury ED model. We examine if human corpora cavernosal smooth muscle responds in a similar manner to SHH treatment. Objective In this study we: 1.) Examine if human corpora cavernosal smooth muscle cells from primary culture of ED patient tissues, respond to Sonic hedgehog (SHH) treatment by increasing growth rate, as in our in vivo animal model. 2.) Determine if human corpora cavernosal cells from patients with differing underlying mechanisms of ED development (prostatectomy, diabetes, hypertension, cardiovascular disease), respond similarly to SHH treatment. 3.) Identify if organ culture conditions (glucose) affect growth of corpora cavernosal smooth muscle and the response to SHH signaling. 4.) Examine the vascular component of ED (hypertension and cardiovascular disease). Methods Human corpora cavernosal tissue was obtained from prostatectomy, diabetic, hypertension, cardiovascular disease and Peyronie's (control) patients (n=40) that were under going prosthesis implant to treat ED. Primary cultures (n=17) were established, and corpora cavernosal cells from passage 3-4 were treated with SHH protein, MSA (control), 5E1 SHH inhibitor, and PBS (control). Growth was quantified by counting the number of cells using a hemocytometer at 3-4 days. The concentration of SHH protein for maximal growth was examined, in addition to a lipid modified more active SHH peptide. Results SHH treatment increased smooth muscle growth in human corpora cavernosal cells from prostatectomy, diabetic, and Peyronie's patients in a similar manner (43-53%). SHH inhibition decreased smooth muscle growth (24-32%). There was no difference in growth using 25ug and 10ug SHH peptide. A more active (150X) SHH peptide further enhanced growth (20%). SHH protein increased growth more in diabetic smooth muscle under high glucose conditions. SHH inhibition was less effective under high glucose conditions. SHH treatment increased growth less in cells from hypertension and cardiovascular disease patients. Conclusion Corpora cavernosa smooth muscle from prostatectomy, diabetic, and Peyronie's patients increased in response to SHH treatment in a similar manner, suggesting that SHH treatment would be beneficial to enhance smooth muscle regeneration in patients with ED of multiple origins. There is a threshold concentration of SHH protein above which smooth muscle growth is enhanced. Lipid modified SHH engendered a more robust growth response. High glucose conditions that may be present in under controlled diabetic patients would not detract from SHH usefulness as a protectant for corpora cavernosal morphology. Understanding how human corpora cavernosal tissue responds to SHH treatment is critical for clinical translation to ED patients. Disclosure Work supported by industry: no.

MP21-02 PREDICTORS OF CORPORO-VENOCCLUSIVE DYSFUNCTION (CVOD) IN MEN WITH BILATERAL NERVE-SPARING RADICAL PROSTATECTOMY (RP)

You have accessJournal of UrologyCME1 May 2022MP21-02 PREDICTORS OF CORPORO-VENOCCLUSIVE DYSFUNCTION (CVOD) IN MEN WITH BILATERAL NERVE-SPARING RADICAL PROSTATECTOMY (RP) Jose M. Flores, Carolyn A. Salter, Nicole Benfante, Elizabeth Schofield, and John P. Mulhall Jose M. FloresJose M. Flores More articles by this author , Carolyn A. SalterCarolyn A. Salter More articles by this author , Nicole BenfanteNicole Benfante More articles by this author , Elizabeth SchofieldElizabeth Schofield More articles by this author , and John P. MulhallJohn P. Mulhall More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002554.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Nerve-sparing status (NSS) during RP is a significant predictor of erectile function recovery (EFR). Poor nerve sparing is associated with collagenization of cavernosal smooth muscle and CVOD development with typically poor response to PDE5 inhibitors (PDE5i) and intracavernosal injection therapy (ICI). This study aimed to define factors predictive of CVOD in men with bilateral NS RP. METHODS: Study population consisted of (i) men who underwent bilateral NS (defined by the surgeon using an institutional grading system) RP (ii) with poor response to PDE5i and ICI (iii) ≥2 years post-RP (iv) who underwent penile duplex Doppler ultrasound (DUS) (v) using a vasoactive agent redosing schedule. CVOD diagnosis was made with end-diastolic velocity (EDV) values ≥5 cm/sec bilaterally. ADT and radiation therapy were exclusions. We used multivariable modeling to define factors predictive of CVOD. Factors included in the model: patient age, history of diabetes, history of obstructive sleep apnea, number of vascular comorbidities, baseline EF, peri-RP total testosterone (T) level, and total T at DUS (within 3 months). RESULTS: 201 men with a mean age of 66 ± 7 years. 25% of the patients reported 3 or more vascular comorbidities, 17% had diabetes, 36% obstructive sleep apnea, 39% were current or former smokers. Mean baseline PSA and pre-RP total T was 6.7 ± 6 ng/mL and 410 ± 211 ng/dL, respectively. Median baseline EFD score was 24 (13, 29). 49% of the men had an erection less than penetration rigidity during the DUS procedure with a mean total dose administered of 87 ± 21 units of trimix. 69% received 100 units. CVOD was diagnosed in 76%. Mean total T at DUS was 386 ± 177 ng/dL. 32% had low T. Significant predictors of CVOD after RP in men with BL NSS were: OSA (OR = 3.8, 95% CI = 1.7-8.7), ≥ 3 vascular comorbidities (OR = 5.0, 95% CI = 1.7-14.8), age at the time of DUS (per 10 years increase, OR = 1.9, 95% CI = 1.2-3.0), and pre-RP EF domain score (per 1 unit increase, OR = 0.93, 95% CI 0.88-0.98). There was a signal that in a larger sample size, diabetes may be a significant predictor of CVOD in this group of patients (OR = 2.7, 95% 0.9-8.0). CONCLUSIONS: Baseline clinical comorbidities pre-RP have a significant impact on EFR post-RP, in particular, the presence of venous leak. Despite BL NSS, older patients, men with OSA, higher number of comorbidities pre-RP, and lower pre-RP EF increased the likelihood of developing CVOD post-RP. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e327 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jose M. Flores More articles by this author Carolyn A. Salter More articles by this author Nicole Benfante More articles by this author Elizabeth Schofield More articles by this author John P. Mulhall More articles by this author Expand All Advertisement PDF DownloadLoading ...

MP21-20 EXPLORING ERECTILE HEMODYNAMICS IN MEN COMPLAINING OF HARD-FLACCID SYNDROME (HFS)

You have accessJournal of UrologyCME1 May 2022MP21-20 EXPLORING ERECTILE HEMODYNAMICS IN MEN COMPLAINING OF HARD-FLACCID SYNDROME (HFS) John Sullivan, Carolyn Salter, Jose Flores, Pat Guhring, Marilyn Parker, and John Mulhall John SullivanJohn Sullivan More articles by this author , Carolyn SalterCarolyn Salter More articles by this author , Jose FloresJose Flores More articles by this author , Pat GuhringPat Guhring More articles by this author , Marilyn ParkerMarilyn Parker More articles by this author , and John MulhallJohn Mulhall More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002554.20AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: There is increasing awareness of a condition, vernacularly termed HFS, in which men complain of subjective and palpable abnormalities in the flaccid penis (hardness, heaviness) often associated with erectile dysfunction (ED). We attempted to define erectile hemodynamic profiles in this population. METHODS: The study population consisted of (i) males ≥18 years of age (ii) who presented with self-diagnosed HFS (iii) who underwent comprehensive history and examination (including genital and lower extremity neurological) and (iv) penile duplex Doppler ultrasound (PDDU). PDDU was performed using redosing of vasodilators. PSV ≥35 cm/s and EDV ≤3 were considered normal. Patients with Peyronie’s disease or a history of intracavernosal injection therapy were excluded. RESULTS: 88 men were analyzed. Mean age was 28±12 years. Median number of vascular comorbidities was zero (0, 1). 66 (75%) had a history of depression or anxiety, 60 of whom had used pharmacotherapy for such at some time and 40 were actively using anxiolytics or anti-depressants at time of evaluation. No patient had a history of neurological conditions and neurological exam was normal in all. Mean duration of symptoms was 14 ± 20 months. Median number of prior physician consultations was 1 (1,4). All complained of impaired erectile function, 50% being incapable of having penetrative sexual relations. 91% had or were using PDE5 inhibitors. On examination all patients had a hypertonic flaccid penis with excellent stretch on traction. At PDDU, median number of penile injections was 2 (1, 3). 50% of men required phenylephrine reversal. All patients had normal PSV with 2/88 having abnormal EDV values, both of whom required reversal. Mean PSV was 45±35 cm/s, mean EDV was 2±11 cm/s. No patient had elevated echogenicity on B-mode scanning. CONCLUSIONS: In this analysis, all men presenting with self-reported HFS had normal hemodynamics and an absence of fibrosis, suggesting the absence of vascular or gross cavernosal smooth muscle structural changes. Source of Funding: none © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e334 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information John Sullivan More articles by this author Carolyn Salter More articles by this author Jose Flores More articles by this author Pat Guhring More articles by this author Marilyn Parker More articles by this author John Mulhall More articles by this author Expand All Advertisement PDF DownloadLoading ...

Sonic Hedgehog Signaling in Corpora Cavernosal Cells from Prostatectomy, Diabetic, Hypertension and Peyronie's Patients with Erectile Dysfunction

ABSTRACT Introduction Erectile dysfunction (ED) treatments are minimally effective in prostatectomy and diabetic patients due to injury to the cavernous nerve. With denervation the critical smooth muscle undergoes apoptosis and the penis becomes fibrotic, thus altering corpora cavernosal architecture. In order to devise novel ED therapies, prevention of corpora cavernosal remodeling is critical. Sonic hedgehog (SHH) treatment by peptide amphiphile (PA) nanofiber hydrogel, suppresses smooth muscle apoptosis and improves erectile function in a rat CN injury ED model. We examine if human corpora cavernosal smooth muscle responds in a similar manner to SHH treatment. Objective In this study we: 1.) Examine if human corpora cavernosal smooth muscle cells from primary culture of ED patient tissues, respond to Sonic hedgehog (SHH) treatment by increasing growth rate, as in our in vivo animal model. 2.) Determine if human corpora cavernosal cells from patients with differing underlying mechanisms of ED development (prostatectomy, diabetes, hypertension, cardiovascular disease), respond similarly to SHH treatment. 3.) Identify if organ culture conditions (glucose) affect growth of corpora cavernosal smooth muscle and the response to SHH signaling. 4.) Examine the vascular component of ED (hypertension and cardiovascular disease). Methods Human corpora cavernosal tissue was obtained from prostatectomy, diabetic, hypertension, cardiovascular disease and Peyronie's (control) patients (n=40) that were under going prosthesis implant to treat ED. Primary cultures (n=17) were established, and corpora cavernosal cells from passage 3-4 were treated with SHH protein, MSA (control), 5E1 SHH inhibitor, and PBS (control). Growth was quantified by counting the number of cells using a hemocytometer at 3-4 days. The concentration of SHH protein for maximal growth was examined, in addition to a lipid modified more active SHH peptide. Results SHH treatment increased smooth muscle growth in human corpora cavernosal cells from prostatectomy, diabetic, and Peyronie's patients in a similar manner (43-53%). SHH inhibition decreased smooth muscle growth (24-32%). There was no difference in growth using 25ug and 10ug SHH peptide. A more active (150X) SHH peptide further enhanced growth (20%). SHH protein increased growth more in diabetic smooth muscle under high glucose conditions. SHH inhibition was less effective under high glucose conditions. SHH treatment increased growth less in cells from hypertension and cardiovascular disease patients. Conclusions Corpora cavernosa smooth muscle from prostatectomy, diabetic, and Peyronie's patients increased in response to SHH treatment in a similar manner, suggesting that SHH treatment would be beneficial to enhance smooth muscle regeneration in patients with ED of multiple origins. There is a threshold concentration of SHH protein above which smooth muscle growth is enhanced. Lipid modified SHH engendered a more robust growth response. High glucose conditions that may be present in under controlled diabetic patients would not detract from SHH usefulness as a protectant for corpora cavernosal morphology. Understanding how human corpora cavernosal tissue responds to SHH treatment is critical for clinical translation to ED patients. Disclosure No

The Potential Role of C-peptide in Sexual and Reproductive Functions in Type 1 Diabetes Mellitus: An Update.

Although hyperglycaemia is known to be the leading cause of diabetic complications, the beneficial effect of optimal glucose control in preventing diabetic complications is still far from being proven. In fact, such complications may not be related to glycaemic control alone. This review summarizes several studies that suggest that a C-peptide deficiency could be new and common pathophysiology for complications in type 1 diabetes, including sexual and reproductive dysfunction. We reviewed in vitro, in vivo, and human studies on the association between C-peptide deficiency or C-peptide replacement therapy and complications in type 1 diabetes. It seems that Cpeptide replacement therapy may interrupt the connection between diabetes and sexual/reproductive dysfunction. The Diabetes Control and Complications Trial suggested that maintaining C-peptide secretion is associated with a reduced incidence of retinopathy, nephropathy, and hypoglycaemia. Risk of vascular, hormonal, and neurologic damage in the structures supplying blood to the penis increases with increasing levels of HbA1. However, several human studies have suggested an association between C-peptide production and hypothalamic/pituitary functions. When exposed to C-peptide, cavernosal smooth muscle cells increase the production of nitric oxide. C-peptide in diabetic rats improves sperm count, sperm motility, testosterone levels, and nerve conduction compared to non-treated diabetic rats. C-peptide deficiency may be involved, at least partially, in the development of several pathological features associated with type 1 diabetes, including sexual/reproductive dysfunction. Preliminary studies have reported that C-peptide administration protects against diabetic microand macrovascular damages as well as sexual/reproductive dysfunction. Therefore, further studies are needed to confirm these promising findings.

Long noncoding RNA H19 promotes the apoptosis of corpus cavernsum smooth muscle cells after cavernosal nerve injury via JNK signalling pathway.

JNK/ Bcl-2/ Bax pathway participates in corpus cavernosal smooth muscle cells apoptosis during early period after cavernosal nerve (CN) crush injury (CNCI). Nevertheless, the regulation mechanisms of long noncoding RNA H19 in apoptosis during early stage after CN injury are still poorly understood. The rats in sham group were not direct injury to the CNs. The rats in CNCI group were performed to bilateral CN crush injury. The ICP/MAP rate and smooth muscle content were significantly lower than that in the sham group. Primary CCSMCs were prepared from the tissues samples after completing erectile function detection. Phosphorylated-JNK level was increased significantly, and the expression of Bax and Bcl-2 was elevated and declined in CNCI group respectively. Except for Bcl-2, the mRNA levels of H19, JNK and Bax were significantly increased in CNCI group. After H19 siRNA transfection, for the mRNA and protein levels, JNK and Bax were declined, while Bcl-2 was enhanced. LncRNA H19 might be involved in regulation of Bcl-2, Bax via JNK signalling pathway in CCSMCs apoptosis after CN injury.