Bilateral Cavernous Nerve Crush Research Articles

(063) Effect of Phosphodiesterase Type 4 Inhibitor on Erectile Function Recovery and Nerve Regeneration After Cavernous Nerve Injury in Rats

Abstract Introduction There is no consensus on the best treatment regimen for recovery of erectile function (EF) after radical prostatectomy (RP). Although many pharmacological interventions can improve EF in rats, there is still a lack of studies evaluating strategies that promote nerve regeneration of the cavernous nerve (CN). Phosphodiesterase type 4 inhibitors (PDE4Is), such as rolipram, have shown a neuroprotective effect after spinal cord injury in mice, facilitating functional recovery. It has also been suggested that the neuroprotective property of sildenafil is probably attributed to its effect on phosphodiesterase type 4. Objective Our aim was to evaluate the role of rolipram and its association with tadalafil in EF recovery and nerve regeneration after bilateral CN crush injury (BCNI) in rats. Methods Thirty-five male Wistar rats were randomly divided into four groups: (i) sham (no CN lesion); (ii) control (no treatment after BCNI); (iii) BCNI + tadalafil; (iv) BCNI + rolipram and (v) BCNI + tadalafil + rolipram. The initial laparotomy was performed and the CN were crushed with a hemostatic clamp. The animals in groups (iii) and (iv) received tadalafil (5mg/kg/d) or rolipram (1mg/kg/d) orally for 14 days, respectively. The animals in group (v) received tadalafil + rolipram at the same doses. Then, the animals were submitted to CN stimulation at frequencies 4, 8 and 16 Hz, with recording of mean arterial pressure (MAP) and intracavernosal pressure (ICP). EF was represented by the measurement of maximum ICP normalized to MAP (ICP/MAP ratio) and total normalized area under the curve (AUC) of ICP. The penises and CN of the animals were sent for structural evaluation with the immunohistochemical markers S100 and Ki-67. Results In the functional analysis, BCNI resulted in worse EF when compared to sham. This was observed when considering the maximum ICP/MAP and when analyzing normalized AUC of ICP for 4Hz and 8Hz. The different treatments were able to increase the ICP/MAP ratio after stimulation with 4Hz (p = 0.04) and 8Hz (p = 0.01). When considering normalized AUC of ICP, the different drugs (tadalafil, rolipram or tadalafil + rolipram) showed a significant improvement in EF in two of the three frequencies analyzed (4Hz and 8Hz) (figure 1). For 16hz, although it did not reach a statistical difference, a trend of improvement was observed after the different treatments (p = 0.08). With regard to immunohistochemical analysis, the tadalafil group had a higher expression of S100 in the dorsal penile nerve compared to the control group (p = 0.01). The rolipram group had a lower expression of Ki-67 in the CN compared to the control group (p = 0.03). Conclusions The combination of tadalafil and rolipram was associated with a functional improvement in EF in the experimental model with rats. The present study also suggests that rolipram may have a neuroprotective effect according to the structural evaluation of the CN. Disclosure No.

Knockdown of programmed cell death factor 4 restores erectile function by attenuating apoptosis in rats with bilateral cavernous nerve crush injury.

Apoptosis is an important pathologic mechanism of erectile dysfunction after radical prostatectomy. Studies have shown that programmed cell death factor 4 is connected to the modulation of apoptosis in many cells. However, the programmed cell death factor 4 function in the cavernous nerve injury erectile dysfunction is unclear. This investigation aimed to explore the programmed cell death factor 4 function in erectile dysfunction in rats with bilateral cavernous nerve crush. The experiment used 30 male Sprague Dawley rats (18 months old) that were screened for normal erectile function by the apomorphine test. Ten rats were randomized into Sham and bilateral cavernous nerve crush groups to detect changes in programmed cell death factor 4 expression. The remaining 20 rats were distributed at random to four groups: the Sham group treated by sham surgery, the phosphate-buffered saline group, the lentivirus containing negative control short hairpin RNA group, and the lentivirus containing short hairpin RNA targeting programmed cell death factor 4 group underwent bilateral cavernous nerve crush and were afterward administered intracavernous injections of phosphate-buffered saline, lentivirus containing negative control short hairpin RNA, or lentivirus containing short hairpin RNA targeting programmed cell death factor 4. Electrical stimulation of the cavernous nerve was conducted 2 weeks later for penile erectile function assessment. The cavernous tissue was collected for histological analysis and western blotting. The apoptosis level in rat corpus cavernosum was elevated, and programmed cell death factor 4 expression was increased after bilateral cavernous nerve crush. Knockdown of programmed cell death factor 4 significantly improved erectile function in bilateral cavernous nerve crush rats. Furthermore, lentivirus containing short hairpin RNA targeting programmed cell death factor 4 treatment raised smooth muscle content and attenuated cavernous fibrosis and apoptotic levels. Additionally, programmed cell death factor 4 was found to mediate the PI3K/AKT pathway. Elevated programmed cell death factor 4 expression may be an important pathogenetic mechanism for erectile dysfunction after bilateral cavernous nerve crush, and the knockdown of programmed cell death factor 4 enhanced erectile function in 18-month-old rats after cavernous nerve damage. The potential mechanism may be the stimulation of the PI3K/AKT pathway to attenuate the cavernous apoptosis level.

Functional recovery of injured cavernous nerves achieved through endogenous nerve growth factor-containing bioactive fibrous membrane.

Radical prostatectomy is a highly successful treatment for prostate cancer, among the most prevalent manifestations of the illness. Damage of the cavernous nerve (CN) during prostatectomy is the main cause of postoperative erectile dysfunction (ED). In this study, the capability of a personalized bioactive fibrous membrane to regenerate injured CN was investigated. The fibrous membrane bioactivity is conferred by the selectively bound nerve growth factor (NGF) present in the rat urine. In a rat model of bilateral CN crush, the implanted bioactive fibrous membrane induces CN regeneration and restoration of erectile function, showing a significantly increased number of smooth muscle cells and content of endothelial and neuronal nitric oxide synthases (eNOS; nNOS). In addition, the bioactive fibrous membrane promotes nerve regeneration by increasing the number of myelinated axons and nNOS-positive cells, therefore reversing the CN fibrosis found in untreated rats or rats treated with a bare fibrous membrane. Therefore, this personalized regenerative strategy could overcome the recognized drawbacks of currently available treatments for CN injuries. It may constitute an effective treatment for prostate cancer patients suffering from ED after being subject to radical prostatectomy. STATEMENT OF SIGNIFICANCE: The present work introduces a unique strategy to address post-surgical ED resulting from CN injury during pelvic surgery (e.g., radical prostatectomy, radical cystoprostatectomy, abdominoperineal resection). It comprises a bioactive and cell-free fibrous implant, customized to enhance CN recovery. Pre-clinical results in a rat model of bilateral CN crush demonstrated that the bioactive fibrous implant can effectively heal injured CN, and restore penile structure and function. This implant selectively binds NGF from patient fluids (i.e. urine) due to its functionalized surface and high surface area. Moreover, its local implantation reduces adverse side effects. This tailored regenerative approach has the potential to revolutionize the treatment of ED in prostate cancer patients following radical prostatectomy, overcoming current treatment limitations.

Molecular mechanisms of penile traction for penile rehabilitation in a bilateral cavernous nerve crush injury rat model.

Prostate cancer is the most common solid-organ malignancy in adult men. Early detection and treatment of prostate cancer with radical prostatectomy (RP) has improved cancer-specific survival but is associated with penile shortening and erectile dysfunction. Penile traction therapy (PTT) has been demonstrated to increase stretched penile length (SPL) prior to penile prosthesis placement and may improve erectile function (EF) in patients with Peyronie's disease. We aimed to evaluate the efficacy of PTT in preserving penile length and EF after bilateral cavernous nerve crush injury (BCNI) in a rat model. Twenty-four male Sprague-Dawley rats aged 11-13 weeks were randomly assigned to three groups (n=8, each): sham operation with no PTT (Sham), BCNI without PTT (Crush), and BCNI with PTT (Traction). PTT was started on postoperative day 3. A traction force of 1 Newton was applied to the penis for 30 minutes each day for 28 days. After 28 days of traction, the cavernous nerve was stimulated while recording the intracavernosal pressure (ICP) and the mean arterial pressure (MAP) simultaneously. Cavernosal tissue was excised, and western blot analysis for endothelial nitric oxide synthase (eNOS) was performed. Significance was determined by using ANOVA with Tukey-Kruger post-hoc testing. At 4 weeks after nerve injury, the Traction group had significantly greater SPL compared to the Sham and Crush groups (30 vs. 28 and 27 mm, respectively). The Sham group had significantly greater EF (ΔICP/MAP) compared to the Crush group at 2.5, 5, and 7.5 V. The EF of the Traction group was between that of the Sham and Crush groups and was not significantly different from the Sham group at any voltages. Further downstream analysis revealed that the Traction group had significantly greater eNOS expression in cavernosal tissue compared to the Crush group, which was confirmed on western blot analysis and immunohistochemistry (IHC) staining. Findings from this animal study suggest that PTT has the potential to mitigate penile retraction after RP. While more studies are needed to determine the effect of PTT on preservation of EF, the increased eNOS expression observed in the Traction group offers a potential protective mechanism of action.

(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 <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 (>2 fold; Q<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

LIPUS-SCs-Exo promotes peripheral nerve regeneration in cavernous nerve crush injury-induced ED rats via PI3K/Akt/FoxO signaling pathway.

Clinical treatment of erectile dysfunction (ED) caused by cavernous nerve (CN) injury during pelvic surgery is difficult. Low-intensity pulsed ultrasound (LIPUS) can be a potential strategy for neurogenic ED (NED). However, whether Schwann cells (SCs) can respond to LIPUS stimulation signals is unclear. This study aims to elucidate the signal transmission between SCs paracrine exosome (Exo) and neurons stimulated by LIPUS, as well as to analyze the role and potential mechanisms of exosomes in CN repair after injury. The major pelvic ganglion (MPG) neurons and MPG/CN explants were stimulated with LIPUS of different energy intensities to explore the appropriate LIPUS energy intensity. The exosomes were isolated and purified from LIPUS-stimulated SCs (LIPUS-SCs-Exo) and non-stimulated SCs (SCs-Exo). The effects of LIPUS-SCs-Exo on neurite outgrowth, erectile function, and cavernous penis histology were identified in bilateral cavernous nerve crush injury (BCNI)-induced ED rats. LIPUS-SCs-Exo group can enhance the axon elongation of MPG/CN and MPG neurons compared to SCs-Exo group in vitro. Then, the LIPUS-SCs-Exo group showed a stronger ability to promote the injured CN regeneration and SCs proliferation compared to the SCs-Exo group in vivo. Furthermore, the LIPUS-SCs-Exo group increased the Max intracavernous pressure (ICP)/mean arterial pressure (MAP), lumen to parenchyma and smooth muscle to collagen ratios compared to the SCs-Exo group in vivo. Additionally, high-throughput sequencing combined with bioinformatics analysis revealed the differential expression of 1689 miRNAs between the SCs-Exo group and the LIPUS-SCs-Exo group. After LIPUS-SCs-Exo treatment, the phosphorylated levels of Phosphatidylinositol 3-kinase (PI3K), protein kinase B (Akt) and forkhead box O (FoxO) in MPG neurons increased significantly compared to negative control (NC) and SCs-Exo groups. Our study revealed that LIPUS stimulation could regulate the gene of MPG neurons by changing miRNAs derived from SCs-Exo, then activating the PI3K-Akt-FoxO signal pathway to enhance nerve regeneration and restore erectile function. This study had important theoretical and practical significance for improving the NED treatment.

MP27-03 RNASEQ AND PATHWAY ANALYSIS OF RAT PENIS WITH CAVERNOUS NERVE INJURY AND SONIC HEDGEHOG TREATMENT BY PEPTIDE AMPHIPHILE

You have accessJournal of UrologyCME1 Apr 2023MP27-03 RNASEQ AND PATHWAY ANALYSIS OF RAT PENIS WITH CAVERNOUS NERVE INJURY AND SONIC HEDGEHOG TREATMENT BY PEPTIDE AMPHIPHILE Timothy Searl, Sarah Martin, Daniel Harrington, Samuel Stupp, Samuel Ohlander, Kevin Mcvary, and Carol Podlasek Timothy SearlTimothy Searl More articles by this author , Sarah MartinSarah Martin More articles by this author , Daniel HarringtonDaniel Harrington More articles by this author , Samuel StuppSamuel Stupp 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.0000000000003255.03AboutPDF 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 (CN). CN injury causes loss of innervation, apoptosis of smooth muscle, fibrosis in the penis and ED. When developing novel therapies, the goal is to prevent the apoptotic response and penile remodeling, to preserve erectile function. Our previous studies found Sonic hedgehog (SHH) is a key mediator of penile remodeling. Our objective was to use RNA seq and pathway analysis, to identify changes in gene expression and signaling pathways that are altered in response to CN injury and the effect of SHH treatment by peptide amphiphile (PA), in a rat prostatectomy model of ED. METHODS: Corpora cavernosal (CC) tissue was obtained from Sprague Dawley rats (n=36) that underwent bilateral CN crush, CN crush plus SHH PA treatment of the penis, or sham surgery. 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 directional pathway analysis was examined with Ingenuity. A 2-fold change with a q-value <0.05 was used as a threshold for differential expression. RESULTS: mRNA representing 523 genes had significant differential expression in CN crushed CC tissue relative to sham. Ingenuity analysis predicted increased necrosis, and decreased 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 associated with CN injury in a rat prostatectomy model. Pathway analysis predicted increases in processes associated with cell death and a loss of functions necessary for maintenance of CC architecture. Most CN injury associated changes were reversed or greatly diminished with SHH PA treatment of the penis at the time of injury. The results support the significant potential of SHH PA as a preventative treatment for ED in prostatectomy patients. Signaling pathways identified with CN injury provide many new avenues for intervention independently and in combination with SHH PA treatment. Source of Funding: DK101536 © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e362 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Timothy Searl More articles by this author Sarah Martin More articles by this author Daniel Harrington More articles by this author Samuel Stupp 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 ...

Vitamin D3 improved erectile function recovery by regulating autophagy and apoptosis in a rat model of cavernous nerve injury.

Vitamin D3 is an important element in improving erectile function. However, the mechanisms of vitamin D3 remain unknown. Thus, we explored the effect of vitamin D3 on erectile function recovery after nerve injury in a rat model and investigated its possible molecular mechanisms. Eighteen male Sprague-Dawley rats were used in this study. The rats were randomly divided into three groups: the control, bilateral cavernous nerve crush (BCNC), and BCNC + vitamin D3 groups. BCNC model was established in rats by surgery. The intracavernosal pressure and the ratio of intracavernosal pressure to mean arterial pressure were utilized to evaluate erectile function. Masson trichrome staining, immunohistochemistry, terminal deoxynucleotidyl transferase-mediated dUTP nick end labelling and western blot analysis were performed on penile tissues to elucidate the molecular mechanism. The results indicated that vitamin D3 alleviated hypoxia and suppressed the fibrosis signalling pathway by upregulating the expression of eNOS (p = 0.001), nNOS (p = 0.018) and α-SMA (p = 0.025) and downregulating the expression of HIF-1α (p = 0.048) and TGF-β1 (p = 0.034) in BCNC rats. Vitamin D3 promoted erectile function restoration by enhancing the autophagy process through decreases in the p-mTOR/mTOR ratio (p = 0.02) and p62 (p = 0.001) expression and increases in Beclin1 expression (p = 0.001) and the LC3B/LC3A ratio (p = 0.041). Vitamin D3 application improved erectile function rehabilitation by suppressing the apoptotic process through decreases in the expression of Bax (p = 0.002) and caspase-3 (p = 0.046) and an increase in the expression of Bcl2 (p = 0.004). Therefore, We concluded that vitamin D3 improved the erectile function recovery in BCNC rats by alleviating hypoxia and fibrosis, enhancing autophagy and inhibiting apoptosis in the corpus cavernosum.

Buyang Huanwu Decoction Ameliorates Damage of Erectile Tissue and Function Following Bilateral Cavernous Nerve Injury.

To verify the effect of Buyang Huanwu Decoction (BHD) in ameliorating erectile dysfunction (ED) after radical prostatectomy (RP). The composition of BHD was verified by ultra-performance liquid chromatography quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS/MS) analysis. Bilateral cavernous nerve crush injury (BCNI) in rats was used to mimic the neurovascular injury occurring after RP. By the envelope method, forty rats were randomly divided into 4 groups as follows: sham (cavernous nerves exposed only), model (BCNI), low-dosage BHD [LBHD, 12.8 g/(kg·d)], and high-dosage BHD [HBHD, 51.2 g/(kg·d)] groups, 10 rats in each group, feeding for 3 weeks respectively. Erectile function was evaluated by measuring intracavernosal pressure (ICP). Changes in the histopathology of corpus cavernosum (CC) were examined by hematoxylin-eosin staining. Meanwhile, the fibrosis of CC was measured by Masson's trichrome staining and Western blot was used to detect the expressions of collagen I, transforming growth factor beta 1 (TGF- β 1) and α-smooth muscle actin (α-SMA). Apoptosis index was detected by terminal-deoxynucleoitidyl transferase mediated nick end labeling (TUNEL) and Western blot for determining the expressions of B-cell lymphoma 2 (Bcl-2) and Bcl-2-associated X (Bax). The oxidative stress in the CC were assessed by the superoxide dismutase (SOD), malondialdehyde (MDA) and reactive oxygen species (ROS) levels. The proteins expression of c-Jun N-terminal kinase (JNK) and c-Jun were detected by Western blot. In addition, the expression of α-SMA and p-c-Jun in the CC was observed by double immunofluorescence staining. The UPLC-QTOF-MS/MS analysis showed that BHD contained calycosin-7-O- β -D-glucoside, ononin, calycosin and formononetin. Compared with the model group, LBHD and HBHD treatment improved the ICP and the circumference, area, and weight of CC (P<0.05 or P<0.01). Furthermore, LBHD and HBHD treatments increased CC smooth muscle content and decreased apoptosis index (P<0.05 or P<0.01). LBHD and HBHD also elevated SOD and expression level of α -SMA and Bcl-2, and reduced MDA and ROS levels, as well as expression of TGF- β 1, collagen I, Bax, p-c-JNK, p-JNK in the CC compared with the model group (P<0.05 or P<0.01). The double immunofluorescence staining showed that the fluorescence degree of p-c-Jun in both LBHD and HBHD treatment groups was significantly reduced, whereas the α -SMA expression increased (P<0.05 or P<0.01). BHD can improve ED of rats with BCNI, which is related to inhibiting fibrosis, apoptosis, and oxidative stress of CC. The ROS/JNK/c-Jun signaling pathway may play an important role in the process.

Relaxin-2 Prevents Erectile Dysfunction by Cavernous Nerve, Endothelial and Histopathological Protection Effects in Rats with Bilateral Cavernous Nerve Injury.

Cavernous nerve injury induced erectile dysfunction (ED) is a refractory complication with high incidence in person under radical prostatectomy. Studies have shown that relaxin-2 (RLX-2) plays a vital role of endothelial protection, vasodilation, anti-fibrosis and neuroprotection in a variety of diseases. However, whether penile cavernous erection can benefit from RLX-2 remains unknown. The purpose of the experiment was to explore the effects of RLX-2 on ED in the rat suffering with bilateral cavernous nerve injury (BCNI). The rats were divided into three groups: Sham group was underwent sham operation, BCNI+RLX group or BCNI group was underwent bilateral cavernous nerve crush and then randomly treated with RLX-2 (0.4 mg/kg/d) or saline by continuous administration using a subcutaneously implanted micro pump for 4 weeks respectively. Then, erectile function was evaluated by electrical stimulation of cavernous nerves. Cavernous nerves and penile tissues and were collected for histological evaluation. Erectile function of rats with BCNI was partially improved after RLX-2 treatment. The BCNI group had lower expression of relaxin family peptide receptor (RXFP) 1, p-AKT/AKT, p-eNOS/eNOS ratios than sham operation rats, but RLX-2 could partially reversed these changes. Histologically, the BCNI+RLX group had a significant effect on preservation of neurofilament, neuronal glial antigen 2 of penile tissue and nNOS of cavernous nerves when compared with BCNI group. RLX-2 could inhibited the lever of BCNI induced corporal fibrosis and apoptosis via regulating TGFβ1-Smad2/3-CTGF pathway and the expression of Bax/Bcl-2 ratio, caspase3. RLX-2 could improve erectile function of BCNI rats by protecting cavernous nerve and endothelial function and suppressing corporal fibrosis and apoptosis via RXFP1 and AKT/eNOS pathway. Our findings may provide a promising treatment for refractory BCNI induced ED.

Effects of Cells Self-aggregation in the Treatment of Neurogenic Erectile Dysfunction With Traditional Single Cell Suspension of Adipose-derived Stem Cells

ObjectiveTo clarify the effects of cellular self-aggregation of adipose-derived stem cells (ADSCs) on erectile function (EF). MethodsA model of neurogenic erectile dysfunction was performed using bilateral cavernous nerve crush injury in rats. ADSCs suspensions (1 × 106/0.2 ml), were administered via intracavernous injection (ICI) after being allowed to shelve for 0 minute (ICI 0) or 60 minutes (ICI 60) in vitro, as well as cell aggregates isolated from ICI 60 (ICI A). The caudal vein injection group (CVI 60) was used to evaluate whether cell self-aggregation was beneficial to EF when introduced into the peripheral circulation. One day after the transplantation, the distribution of cells was observed. EF and histopathological changes were evaluated after 4 weeks. ResultsApproximately 85% of ADSCs self-aggregated into cell clusters at 60 minutes. The ICI 60 had more significant improvements in EF and more visualized ADSCs retained in the corpus cavernosum (CC) than ICI 0 and CVI 60 (P <.05), but no significant difference between ICI 60 and ICI A. In the CVI 60 group, the cell clusters formed by self-aggregation could hardly reach the CC and were mostly found in lung tissue. Immunofluorescence staining showed increased the content of expressing biomarkers of smooth muscle, nerve within the CC tissue in the ICI groups when compared to the CVI group. ConclusionADSCs self-aggregation before ICI may be an influential factor in the treatment of neurogenic erectile dysfunction. Its potential mechanism may be through improving cell retention in the CC.

PD62-11 THERAPEUTIC EFFECT OF LIPOSOMAL DELIVERY OF HYDROGEN SULFIDE AND XENON ON ERECTILE FUNCTION AFTER CAVERNOUS NERVE INJURY

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology (PD62)1 Sep 2021PD62-11 THERAPEUTIC EFFECT OF LIPOSOMAL DELIVERY OF HYDROGEN SULFIDE AND XENON ON ERECTILE FUNCTION AFTER CAVERNOUS NERVE INJURY Baibing Yang, Dustin Luse, Song Gao, Shaolin Huang, Yong-jian Geng, and Run Wang Baibing YangBaibing Yang More articles by this author , Dustin LuseDustin Luse More articles by this author , Song GaoSong Gao More articles by this author , Shaolin HuangShaolin Huang More articles by this author , Yong-jian GengYong-jian Geng More articles by this author , and Run WangRun Wang More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002099.11AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Erectile dysfunction (ED) is a neurovascular disorder mediated by dysfunction of the nitric oxide/cyclic guanosine monophosphate (NO/cGMP) pathway, and poor relaxation of corpus cavernosum smooth muscle. The two gaseous molecules, hydrogen sulfide (H2S) and xenon (Xe), have been shown to exert neurovascular protective effects in the setting of ischemic and traumatic injuries. Administration of H2S and Xe has been shown to reduce infarct sizes and improve function in animal models of cerebral ischemia–reperfusion injury. The current study tested the hypothesis that to a similar extent, H2S and Xe both can preserve erectile function after cavernous nerve injury. METHODS: A model of cavernous nerve injury ED was employed using bilateral cavernous nerve crush (BCNC) on adult male Sprague Dawley rats. The experimental and control groups were BCNC + saline, BCNC + H2S, BCNC + Xe treatment and a sham surgery control group. H2S treatment was used as positive control, which has been reported to improve erectile function through smooth muscle relaxation. H2S and Xe were packed in echogenic liposomes (ELIPs, liposome diameter ranged from 800 nm to several microns) respectively and injected through the rat tail vein (5mg ELIPs). Gas was released from ELIPs under ultrasound exposure at the root of penis. Intracavernosal pressure (ICP) and mean arterial pressure (MAP) were examined after ultrasound exposure and recorded for all groups. RESULTS: Rats injured by BCNC showed erectile dysfunction evidence by reduced ICP/MAP. Treatment with H2S and Xe blocked the BCNC adverse impact on erectile function by 1.83- and 1.75-fold increases respectively when compared with BCNC group. CONCLUSIONS: Liposomal delivery of H2S and Xe demonstrates potential as a novel therapy for ED after cavernous nerve injury. Source of Funding: None © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1076-e1076 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Baibing Yang More articles by this author Dustin Luse More articles by this author Song Gao More articles by this author Shaolin Huang More articles by this author Yong-jian Geng More articles by this author Run Wang More articles by this author Expand All Advertisement Loading ...

PD20-02 PENILE TRACTION THERAPY INCREASES STRETCHED PENILE LENGTH AND ENDOTHELIAL NITRIC OXIDE SYNTHASE EXPRESSION IN A BILATERAL CAVERNOUS NERVE CRUSH INJURY RAT MODEL

You have accessJournal of UrologySexual Function/Dysfunction: Medical, Hormonal & Non-surgical Therapy I (PD20)1 Sep 2021PD20-02 PENILE TRACTION THERAPY INCREASES STRETCHED PENILE LENGTH AND ENDOTHELIAL NITRIC OXIDE SYNTHASE EXPRESSION IN A BILATERAL CAVERNOUS NERVE CRUSH INJURY RAT MODEL Brian Dick, Jacob Greenberg, Michael Polchert, Max Moore, Joseph Kim, Cameron Belding, Hogyoung Kim, Suresh Sikka, Asim Abdel-Mageed, Shams Halat, and Wayne Hellstrom Brian DickBrian Dick More articles by this author , Jacob GreenbergJacob Greenberg More articles by this author , Michael PolchertMichael Polchert More articles by this author , Max MooreMax Moore More articles by this author , Joseph KimJoseph Kim More articles by this author , Cameron BeldingCameron Belding More articles by this author , Hogyoung KimHogyoung Kim More articles by this author , Suresh SikkaSuresh Sikka More articles by this author , Asim Abdel-MageedAsim Abdel-Mageed More articles by this author , Shams HalatShams Halat More articles by this author , and Wayne HellstromWayne Hellstrom More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002009.02AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Prostate cancer is the most common type of solid-organ cancer in men. Early detection and treatment with radical prostatectomy has improved cancer-specific survival but is associated with penile shortening and erectile dysfunction. Penile traction therapy (PTT) has been demonstrated to increase stretched penile length (SPL) prior to penile prosthesis placement, and may improve erectile function in patients with Peyronie’s Disease. We aim to evaluate the efficacy of PTT in preserving penile length and erectile function after bilateral cavernous nerve crush injury (BCNI) in a rat model. METHODS: 24 Male Sprague-Dawley rats aged 11-13 weeks were randomly assigned to 3 groups (n=8, each): sham operation with no PTT (sham); BCNI without PTT (crush); and BCNI with PTT (traction). PTT was started on post-op day 3. A traction force of 1 Newton was applied to the penis for 30 minutes each day, for 28 days. Penis measurements were taken every 7 days. After 28 days of traction, the cavernous nerve was stimulated while recording mean arterial pressure (MAP) and intracavernosal pressure (ICP). Cavernosal tissue was excised and Western blot analysis for endothelial nitric oxide synthase (eNOS) was performed. Significance was determined using ANOVA and Tukey’s test. RESULTS: At 4 weeks after nerve injury, animals treated with traction therapy had significantly greater SPL compared to sham and crush groups (30.0mm vs 27.9mm and 27.1mm respectively). Animals who received a sham operation had significantly greater erectile function (ΔICP/MAP) compared to the crush group at all voltages (2.5V P <0.05, 5V P <0.05, and 7.5V P <0.05). There was no significant difference in erectile function between sham and traction or traction and crush at any voltage. The traction group had significantly greater eNOS expression in cavernosal tissue compared to the crush group. CONCLUSIONS: Findings from this animal study suggest that PTT has the potential to mitigate penile retraction after prostatectomy. While more studies are needed to determine the effect of PTT on preservation of erectile function, the increased eNOS expression observed in the traction group offers a potential protective mechanism of action. Source of Funding: SMSNA © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e367-e367 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Brian Dick More articles by this author Jacob Greenberg More articles by this author Michael Polchert More articles by this author Max Moore More articles by this author Joseph Kim More articles by this author Cameron Belding More articles by this author Hogyoung Kim More articles by this author Suresh Sikka More articles by this author Asim Abdel-Mageed More articles by this author Shams Halat More articles by this author Wayne Hellstrom More articles by this author Expand All Advertisement Loading ...

CXCL5 Cytokine Is a Major Factor in Platelet-Rich Plasma's Preservation of Erectile Function in Rats After Bilateral Cavernous Nerve Injury

BackgroundThe neuro-protective and tissue-protective properties of platelet-rich plasma (PRP) have been demonstrated through treating bilateral cavernous nerve (CN) injury in rats, although the underlying mechanisms have not been fully clarified. AimTo determine factors released from PRP and explore their role in mediating preservation of erectile function (EF) in a rat model of CN injury. MethodsMale Sprague-Dawley rats (aged 10 weeks) were used in this study. 6 rats were used to obtain blood for PRP and whole plasma preparation. We probed samples using a cytokine antibody array and performed enzyme-linked immunosorbent assay (ELISA). We determined the expression patterns of C-X-C motif chemokine ligand 5 (CXCL5) and receptors in the major pelvic ganglion (MPG) and corpus cavernosum via immunostaining. 32 rats were divided into 4 groups based on the type of injection received: (i) sham, (ii) vehicle, (iii) 400 μL of PRP, and (iv) 30 ng/kg of CXCL5. Groups 2–4 were subjected to bilateral CN crush (BCNC) injury. 4 weeks later, EF was assessed by CN electrostimulation, and CNs and penile tissue were collected for histological analysis. OutcomeCytokine antibody array, ELISA, erectile response, and immunofluorescence staining readings. ResultsThe PRP contained high levels of CXCL5. MPG neurons expressed CXCL5 and CXCR2. PRP intracavernous injection stabilized CXCR2 and increased CXCL5 expression in the MPG after BCNC, thus enhancing neuroprotection. CXCL5 injection improved BCNC-induced erectile dysfunction by preventing smooth muscle atrophy. Clinical ImplicationsThe therapeutic efficacy of PRP in CN injury–induced erectile dysfunction may arise from the synergy among multiple biomolecules. Our study serves as a basis for future studies on PRP formulation to provide safe and effective medications for the maintenance of EF after radical prostatectomy in patients with prostate cancer. Strengths & LimitationsA strength of our study is that our model was able to isolate the role of cytokines, specifically CXCL5, as part of the mechanism responsible for PRP's protective properties. However, the rat cytokine array provided limited experimental targets. The rats used were not at the age corresponding to prostate cancer patients in clinical settings. Our study did not explore CXCL5 blocking in the PRP group. Finally, the main protein quantification results by western blotting were hampered because of small tissue samples. ConclusionsThis study provides evidence for the role of CXCL5 and CXCR2 as mediators of PRP effects in the preservation of EF after CN injury.Wu YN, Liao CH, Chen KC, et al. CXCL5 Cytokine Is a Major Factor in Platelet-Rich Plasma's Preservation of Erectile Function in Rats After Bilateral Cavernous Nerve Injury. J Sex Med 2021;18:698–710.

086 Penile Traction Therapy Improves Penile Length and Increases Cavernosal Endothelial Nitric Oxide Synthase Expression in a Rat Model of Bilateral Cavernous Nerve Crush Injury

Prostate cancer is the most common type of solid-organ cancer in men. Early detection and treatment with radical prostatectomy has improved cancer-specific survival but is associated with penile shortening and erectile dysfunction. Penile traction therapy (PTT) has been shown to increase penile length prior to penile prosthesis placement, and there is preliminary data suggesting that it improves erectile function in patients with Peyronie’s Disease.

Vacuum therapy ameliorates erectile dysfunction in bilateral cavernous nerve crush rats by inhibiting apoptosis and activating autophagy.

Postprostatectomy erectile dysfunction (pPED) remains a current problem despite improvements in surgical techniques. Vacuum therapy is clinically confirmed as a type of pPED rehabilitation. However, its underlying mechanisms are incompletely understood. Recently, autophagy and apoptosis were extensively studied in erectile dysfunction resulting from diabetes, senescence, and androgen deprivation but not in the context of pPED and vacuum therapy. Therefore, this study was designed to investigate the roles of autophagy and apoptosis in pPED and vacuum therapy. Twenty-four adult male Sprague–Dawley rats were randomly divided into three groups: the control group, bilateral cavernous nerve crush (BCNC) group, and BCNC + vacuum group. After 4 weeks of treatment, intracavernosal pressure was used to evaluate erectile function. Real-time quantitative polymerase chain reaction, western blot, and immunohistochemistry were used to measure the molecular expression. TdT-mediated dUTP nick-end labeling staining was used to assess apoptosis. Transmission electron microscopy was used to observe autophagosomes. After treatment, compared with those of the BCNC group, erectile function and cavernosal hypoxia had statistically significantly improved (P < 0.05). Apoptosis and the relative protein expression of B-cell lymphoma-2-associated X and cleaved Caspase3 were decreased (P < 0.05). Autophagy-related molecules such as phosphorylated unc-51-like autophagy-activating kinase 1 (Ser757) and p62 were decreased. Beclin1, microtubule-associated protein 1 light chain 3 A/B, and autophagosomes were increased (P < 0.05). Besides, the phosphatidylinositol 3-kinase/AKT/mammalian target of rapamycin signaling pathway, as a negative regulator of autophagy to some degree, was inhibited. This study revealed that vacuum therapy ameliorated pPED in BCNC rats by inhibiting apoptosis and activating autophagy.

Whole-transcriptome analysis of rat cavernosum and identification of circRNA-miRNA-mRNA networks to investigate nerve injury erectile dysfunction pathogenesis

ABSTRACT There is growing evidence that circular RNAs (circRNAs) play a vital role in many kinds of diseases, including erectile dysfunction (ED). Nevertheless, the role of circRNAs in cavernous nerve-damaging ED (CNI-ED) is unknown. Here, we aimed to discover novel circRNAs, probed their potential role in the CNI-ED, and construct a ceRNA network of circRNAs. Twelve male Sprague Dawley rats were randomly divided into 2 groups by us: bilateral cavernous nerve crush (BCNC) and control groups. Four weeks after surgery, the spongy smooth muscle tissue of the rat penis was sequenced using high-throughput full transcriptome sequencing. We analyzed the expression of circRNAs, miRNAs, and mRNAs in the two groups. Twenty circRNAs with significantly different expressions were selected for RT-qPCR. CeRNA network of circRNAs was established using Cytoscape. GO and KEGG analysis was done by R package. Sequencing showed that 4,587 circRNAs, 762 miRNAs, and 21,661 mRNAs were dysregulated in the BCNC group. The top 20 differentially expressed circRNAs were further verified via RT-qPCR. The ceRNA network contained ten circRNAs, six miRNAs, and 227 mRNAs, including 23 circRNA-miRNA pairs and 227 miRNA-mRNA pairs. GO and KEGG analysis suggested that these ten circRNAs could main regulate energy metabolism processes. A protein‐protein interaction network was constructed with the mRNAs in ceRNA network, and five hub genes were identified. Our study revealed a potential link between circRNAs, miRNAs, and mRNAs in CNI-ED, suggesting that circRNAs may contribute to the occurrence of ED by regulating the cellular energy metabolism in CNI-ED.

Comprehensive Analysis of lncRNA Expression Pattern and lncRNA–miRNA–mRNA Network in a Rat Model With Cavernous Nerve Injury Erectile Dysfunction

BackgroundLong noncoding RNAs (lncRNAs) are differentially expressed in erectile dysfunction (ED) associated with aging and diabetes mellitus; however, the lncRNA expression profile in cavernous nerve (CN) injury–related ED (CNI-ED) is unknown. AimTo investigate the dysregulated lncRNAs, microRNAs (miRNAs), and mRNA expression in CNI-ED and construct a potential lncRNA–miRNA–mRNA network. Methods22 male Sprague–Dawley (SD) rats were divided into bilateral CN crush (BCNC) and Sham groups. Using second-generation high-throughput sequencing technology, we analyzed the expression profiles of lncRNA, miRNA, and mRNA of the 2 groups. 17 differentially expressed lncRNAs were selected and further validated by quantitative real-time polymerase chain reaction (RT-qPCR). The lncRNA–miRNA–mRNA network, Gene Ontology (GO) term enrichment, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses were performed using Cytoscape. OutcomesIntra-cavernosal pressure, mean arterial pressure, smooth muscle content, and the expression of miRNA, mRNA, and lncRNA were measured. ResultsThe BCNC group showed decreased intra-cavernosal/mean arterial pressure as well as decreased smooth muscle/collagen ratios compared with the Sham group. The RNA sequencing results revealed dysregulated expressions of 65 lncRNA, 14 miRNA, and 750 mRNA in the BCNC group based on the following criteria: fold change >2 and P < .05. Among the 17 lncRNAs further selected based on mean count number >4 in both groups, 3 lncRNAs (TCONS_00028173, TCONS_00049985, and TCONS_00058429) were further validated for differential expression by RT-qPCR. GO analysis suggests that these 3 lncRNAs could regulate various processes such as myotube differentiation and muscle cell differentiation. Furthermore, the KEGG pathway analysis showed that the mRNAs in the competing endogenous RNA (ceRNA) network are involved in pathways, including axon guidance and vascular endothelial growth factor signaling pathway. Clinical TranslationOur findings may provide new information on molecular pathophysiology of CNI-ED and suggest further research to find a more effective therapy for CNI-ED. Strengths & LimitationsThis study is the first to identify the lncRNA expression pattern and propose a ceRNA network in a rat model with cavernous nerve injury–related erectile dysfunction. However, analogous studies are needed to confirm these findings in humans. In addition, we constructed the network by only confirming the lncRNA. ConclusionOur study reveals differential expression profiles of lncRNAs, miRNAs, and mRNAs between the BCNC and Sham groups and suggests that these differentially expressed lncRNAs may play critical roles in CNI-ED by regulating apoptosis and fibrosis in the corpus cavernosum via targeting mRNAs or miRNAs.Cong R, Wang Y, Wang Y. Comprehensive Analysis of lncRNA Expression Pattern and lncRNA–miRNA–mRNA Network in a Rat Model With Cavernous Nerve Injury Erectile Dysfunction. J Sex Med 2020;17:1603–1617.

N-acetylcysteine maintains penile length and erectile function in bilateral cavernous nerve crush rat model by reducing penile fibrosis.

Penile length shortening and erectile dysfunction are common complications after radical prostatectomy. Various methods have been used to maintain erectile function, but less attention has been paid to preserving penis length. N-acetylcysteine (NAC) has the effect of antioxidation and antifibrotic, which may be beneficial to improve those postoperative complications. This study investigated the effect of NAC on maintaining the penile length and the erectile function after bilateral cavernous nerve crush (BCNC) and its underlying mechanism. Twenty-four male rats were randomly divided into three groups: control group, BCNC group, and BCNC + NAC group. NAC or equal volume of saline was daily administrated by intragastric gavage for 4 weeks. The initial and end penile lengths were measured. Intracavernosal pressure/mean arterial pressure (ICP/MAP) ratio was calculated to assess erectile function. Hematoxylin–eosin staining, Masson's trichrome staining, immunohistochemistry, and Western blot were performed to explore cellular and molecular changes of the penis. Compared to the BCNC group, the penile length, ICP/MAP ratio and smooth muscle/collagen ratio in the BCNC + NAC group were improved significantly (all P < 0.05), and the expressions of endothelial nitric oxide synthase, α-smooth muscle actin, glutathione, and glutathione peroxidase 1 were significantly increased after NAC treated (all P < 0.05), along with the decreased expressions of hypoxia-inducible factor-1α, transforming growth factor-β1, collagen I, collagen III, collagen IV, malonaldehyde, and lysine oxidase (all P < 0.05). This study demonstrated that NAC could maintain penile length and partly improve erectile function. Possible mechanism is directly and/or indirectly related to antihypoxic and antifibrosis.

Smooth Muscle Progenitor Cells Preserve the Erectile Function by Reducing Corporal Smooth Muscle Cell Apoptosis after Bilateral Cavernous Nerve Crush Injury in Rats.

Radical prostatectomy causes erectile dysfunction (ED) and irreversible morphologic changes, including induction of endothelial and smooth muscle cell (SMC) apoptosis in the corpus cavernosum (CC). The injection of smooth muscle progenitor cells (SPCs) thickens the vascular intima and has demonstrated therapeutic benefit in cardiovascular disease animal. Herein, we investigated the effect of SPCs on the recovery of erectile function (EF) in rat models with bilateral cavernous nerve (CN) injury. Twenty-four male Sprague-Dawley rats were randomized into sham, vehicle only, or SPC treatment groups. Rats in the SPC treatment and vehicle groups were subjected to bilateral CN injury before intracavernosal injection. Intracavernosal injections of SPCs increased all EF parameters at day 28 after injury and simultaneously reduced apoptosis of the SMCs. Ultrastructural analysis revealed that SPCs maintained the integrity of the CC by preserving the structure of the adherens junctions. Tracking transplanted SPCs labeled with EdU showed that transplanted SPCs remained in the CC 28 days after treatment. Intracavernosal SPC injection restored EF after bilateral CN injury by reducing SMC apoptosis, which favored the maintenance of the structure of adherens junctions and regulated the stability of corporal vessels. These findings demonstrate the therapeutic potential of SPCs for treating ED in humans.