Age-related Erectile Dysfunction Research Articles

MicroRNA-145 Gene Modification Enhances the Retention of Bone Marrow-Derived Mesenchymal Stem Cells within Corpus Cavernosum by Targeting Krüppel-Like Factor 4.

The poor retention and ambiguous differentiation of stem cells (SCs) within corpus cavernosum (CC) limit the cell application in erectile dysfunction (ED). Herein, the effects and mechanism of microRNA-145 (miR-145) gene modification on modulating the traits and fate of bone marrow-derived mesenchymal stem cells (BMSCs) were investigated. The effects of miR-145 on cell apoptosis, proliferation, migration, and differentiation were determined by flow cytometry, cell counting kit-8, transwell assays and myogenic induction. Then, the age-related ED rats were recruited to four groups including phosphate buffer saline, BMSC, vector-BMSC, overexpressed-miR-145-BMSC groups. After cell transplantation, the CC were harvested and prepared to demonstrate the retention and differentiation of BMSCs by immunofluorescent staining. Then, the target of miR-145 was verified by quantitative real-time polymerase chain reaction and immunohistochemical. After that, APTO-253, as an inducer of Krüppel-like factor 4 (KLF4), was introduced for rescue experiments in corpus cavernosum smooth muscle cells (CCSMCs) under the co-culture system. In vitro, miR-145 inhibited the migration and apoptosis of BMSCs and promoted the differentiation of BMSCs into smooth muscle-like cells with stronger contractility. In vivo, the amount of 5-ethynyl-2'-deoxyuridine (EdU)+cells within CC was significantly enhanced and maintained in the miR-145 gene modified BMSC group. The EdU/CD31 co-staning was detected, however, no co-staining of EdU/α-actin was observed. Furthermore, miR-145, which secreted from the gene modified BMSCs, dampened the expression of KLF4. However, the effects of miR-145 on CCSMCs could be rescued by APTO-253. Overall, miR-145 modification prolongs the retention of the transplanted BMSCs within the CC, and this effect might be attributed to the modulation of the miR-145/KLF4 axis. Consequently, our findings offer a promising and innovative strategy to enhance the local stem cell-based treatments.

Therapeutic potential of mesenchymal stem cell-derived exosomal miR-296-5p and miR-337-3p in age-related erectile dysfunction via regulating PTEN/PI3K/AKT pathway

Mesenchymal stem cells (MSCs) are viewed as an increasingly promising treatment for age-related erectile dysfunction (AED). Owing to the limitations of injecting living cells, the injection of exosomes appears to be a more plausible option. However, whether MSC-derived exosomes (MSC-Exos) improve AED and their potential mechanism remains unknown. MSC-Exos were prepared and injected intracavernously into aged rats to determine their effects on AED. Masson's trichrome staining was used to ascertain the changes in the histological structure of the corpus cavernosum. Then miRNA sequencing of MSC-Exos and analysis of the critical exosomal miRNAs were performed, as well as their target pathway enrichment analysis. Real-time quantitative PCR (RT-qPCR) and Western blot assay were performed to reveal the functions of MSC-Exos in regulating the PTEN/PI3K/AKT signaling pathway. Moreover, the effects of MSC-Exos on the corpus cavernosum smooth muscle cells (CCSMCs) apoptosis are explored in vitro. The experimental data validate that intracavernous injection of MSC-Exos ameliorated erectile function in AED rats. Masson's trichrome staining shows MSC-Exos therapy restores the histological structure of the corpus cavernosum by improving the ratios of smooth muscle to collagen. The exosomal miR-296–5p and miR-337–3p target and inhibit PTEN, modulating the PI3K/AKT signaling pathway. Furthermore, exosomes inhibit the apoptosis of CCSMCs. Our findings suggest that MSC-Exos improve AED by delivering miR-296–5p and miR-337–3p to regulate the PTEN/PI3K/AKT signaling pathway. These results bode well for the therapeutic potential of MSC-Exos for AED treatment.

Study on the mechanism of aging-related erectile dysfunction based on bioinformatics and experimental verification.

The incidence of aging-related erectile dysfunction (ED) remains high in the elderly population, and has attracted the attention of the medical community. However, aging-related ED responds poorly to traditional treatments for ED, and its mechanism has not yet been fully clarified. This study sought to explore the potential mechanisms of aging-related ED based on bioinformatics and experimental verification. A bioinformatics analysis was performed on data from the Gene Expression Omnibus database related to ED and aging, and the associated differentially expressed genes (DEGs) and signaling pathways were identified. Young and aged rats (n=8 per group) were included in the experimental verification study. Erectile function was detected by electrical stimulation of the cavernous nerve. The corpus cavernosum was collected for the follow-up experiments. A total of 4 hub genes were identified, among which biglycan (BGN) appears to play an important role. The functional enrichment analysis revealed that the extracellular matrix (ECM), especially collagen, related pathways, and the phosphatidylinositol 3‑kinase (PI3K)/protein kinase B (AKT) pathway were enriched, which was also confirmed by the animal experiments. Impaired erectile function in aged rats was associated with the downregulation of the PI3K/AKT pathway, endothelial dysfunction, and increased fibrosis in the penis. Erectile function is impaired with aging. The downregulation of the PI3K/AKT pathway, endothelial dysfunction, and increased fibrosis are involved in this process. BGN may be the key gene regulating these changes. Our study extended understandings of the mechanisms of age-related ED and provides new potential treatment ideas.

Comparative Transcriptome Analyses of Geriatric Rats Associate Age-Related Erectile Dysfunction With a lncRNA-miRNA-mRNA Regulatory Network.

BackgroundThe key regulatory roles of long non-coding RNAs (lncRNAs) in age-related erectile dysfunction (A-ED) are unknown.AimThis study aimed to identify putative lncRNAs that regulate age-related erectile dysfunction via transcriptome analyses, and to predict their specific regulatory routes via bioinformatics methods.Methods22 geriatric male SD rats were divided into age-related erectile dysfunction (A-ED) and negative control (NC) groups after evaluations of intracavernous pressure (ICP). By comparative analysis of transcriptomes of cavernosal tissues from both groups, we identified differentially expressed lncRNAs, miRNAs, and mRNAs. Seven differentially expressed lncRNAs were selected and further verified by quantitative real-time polymerase chain reactions (RT-qPCR). The construction of the lncRNA-miRNA-mRNA network, the Gene Ontology (GO) term enrichment, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were performed in Cytoscape.ResultsFrom comparative transcriptome analyses of A-ED and NC groups, 69, 29, and 364 differentially expressed lncRNAs, miRNAs, and mRNAs were identified respectively. Differentially expressed lncRNAs were culled to seven, which were all verified by qPCR. Three of these lncRNAs (ENSRNOT00000090050, ENSRNOT00000076482, and ENSRNOT00000029245) were used to build regulatory networks, of which only ENSRNOT00000029245 was successful. Moreover, GO and KEGG analyses demonstrated that these lncRNAs possibly regulated muscle myosin complex, muscle cell cellular homeostasis, and ultimately erectile function in rats through PI3K-Akt, fluid shear stress, and atherosclerosis pathways.ConclusionOur study identified differentially expressed lncRNAs, miRNAs, and mRNAs through comparisons of transcriptomes of geriatric rats. An identified lncRNA verified by qPCR, was used to construct a lncRNA-miRNA-mRNA regulatory network. LncRNA ENSRNOT00000029245 possibly regulated downstream mRNAs through this regulatory network, leading to apoptosis in the cavernous tissue, fibrosis, and endothelial dysfunction, which ultimately caused ED. These findings provide seminal insights into the molecular biology of aging-related ED, which could spur the development of effective therapeutics.

Upregulated IGFBP3 with Aging Is Involved in Modulating Apoptosis, Oxidative Stress, and Fibrosis: A Target of Age-Related Erectile Dysfunction.

Aging has been deemed the primary factor in erectile dysfunction (ED). Herein, age-related changes in the erectile response and histomorphology were detected, and the relationship between aging and ED was investigated based on gene expression levels. Thirty male Sprague–Dawley (SD) rats were randomly divided into 6 groups, and intracavernous pressure (ICP) and mean arterial pressure (MAP) were measured. Subsequently, the corpus cavernosum (CC) was harvested and prepared for histological examinations of apoptosis, oxidative stress (OS), and fibrosis. Then, the microarray dataset (GSE10804) was analyzed to identify differentially expressed genes (DEGs) in ED progression, and hub genes were selected. In addition, aged CC smooth muscle cells (CCSMCs) were isolated to evaluate the function of the hub gene by siRNA interference, qRT–PCR, immunofluorescence staining, enzyme-linked immunosorbent assay, western blot analysis, CCK-8 assay, EdU staining, and flow cytometry approaches. The ICP/MAP and smooth muscle cell (SMC)/collagen ratios declined with aging, while apoptosis and OS levels increased with aging. The enriched functions and pathways of the DEGs were investigated, and 15 hub genes were identified, among which IGFBP3 was significantly upregulated. The IGFBP3 upregulation was verified in the CC of aging rats. Furthermore, aged CCSMCs were transfected with siRNA to knock down IGFBP3 expression. The viability and proliferation of the CCSMCs increased, while apoptosis, OS, and fibrosis decreased. Our findings demonstrate that the erectile response of SD rats declines in parallel with enhanced CC apoptosis, OS, and fibrosis with aging. Upregulation of IGFBP3 plays an important role; furthermore, downregulation of IGFBP3 improves the viability and proliferation of CCSMCs and alleviates apoptosis, OS, and fibrosis. Thus, IGFBP3 is a potential therapeutic target for age-related ED.

Modulation of SIRT1 expression improves erectile function in aged rats.

Silent information regulator 2-related enzyme 1 (SIRT1) is an aging-related protein activated with aging. Herein, we evaluated the role of SIRT1 in aging-related erectile dysfunction. The expression of SIRT1 was modulated in aged Sprague-Dawley rats following intragastric administration of resveratrol (Res; 5 mg kg-1), niacinamide (NAM; 500 mg kg-1) or Res (5 mg kg-1) + tadalafil (Tad; phosphodiesterase-5 [PDE5] inhibitor; 5 mg kg-1) for 8 weeks. Then, we determined erectile function by the ratio of intracavernosal pressure (ICP)/mean systemic arterial pressure (MAP). Cavernosal tissues were extracted to evaluate histological changes, cell apoptosis, nitric oxide (NO)/cyclic guanosine monophosphate (cGMP), the superoxide dismutase (SOD)/3,4-methylenedioxyamphetamine (MDA) level, and the expression of SIRT1, p53, and forkhead box O3 (FOXO3a) using immunohistochemistry, terminal deoxynucleotidyl transferase (TdT)-mediated 2'-deoxyuridine 5'-triphosphate (dUTP) nick-end labeling (TUNEL), enzyme-linked immunosorbent assays, and western blot analysis. Compared with the control, Res treatment significantly improved erectile function, reflected by an increased content of smooth muscle and endothelium, NO/cGMP and SOD activity, and reduced cell apoptosis and MDA levels. The effect of Res was improved by adding Tad. In addition, the protein expression of SIRT1 was increased in the Res group, accompanied by decreased p53 and FOXO3a levels. In addition, inhibition of SIRT1 by NAM treatment resulted in adverse results compared with Res treatment. SIRT1 activation ameliorated aging-related erectile dysfunction, supporting the potential of SIRT1 as a target for erectile dysfunction treatment.

Characterization of the Renin-Angiotensin System in Aged Cavernosal Tissue and its Role in Penile Fibrosis

BackgroundThe renin-angiotensin system (RAS) plays an important role in erectile function. The RAS contains 2 major axes: one deleterious, composed of ACE-Ang II-AT1 receptor, and another protective, composed of ACE2-Ang-(1-7)-Mas receptor. While aging is a well-known cause for development of male sexual disorders, little is known about local regulation of the RAS in age-related erectile dysfunction (ED). AimThe present study aimed to assess regulation of the RAS in aging-associated ED rat model and evaluate possible options for disease management through pharmacological modulation of the RAS. MethodsPenile tissues were harvested from 3-, 12-, and 24-month-old Wistar rats. Local expression of major RAS components and ED markers was measured by RT-PCR. Protein expression of RAS components was assessed by western blot. Collagen deposition was measured by Sirius Red and immunohistochemical staining. Evaluation of collagen content was also performed in penile sections of Mas-knockout mice by Sirius Red and Masson’s trichrome stainings. Finally, the effect of Ang-(1-7) pretreatment on TGF-β–induced myofibroblast activation was studied in primary cavernosal and immortalized fibroblasts. OutcomesExperimental results highlighted the essential role of the RAS in modulation of cavernosal fibrosis. ResultsThe present study demonstrates local expression of angiotensinogen mRNA alongside with major RAS components, which suggests local autonomous functioning of the RAS within penile tissue. Gene expression analysis revealed strong positive correlation between ACE-Ang II-AT1 axis with markers for inflammation and fibrosis. While corpus cavernosum from 24-month-old rats was characterized by increased collagen deposition, protein expression of ACE, AT1, and Mas was shown to be upregulated in the penile tissue of this group. At the same time, penile sections from Mas-knockout mice (FVB/N background) were also shown to have increased collagen deposition. Finally, it was demonstrated that Ang-(1-7) treatment of primary cavernosal and immortalized fibroblasts was able to alleviate TGF-β–induced fibroblast-to-myofibroblast transition. Clinical TranslationThe present study suggests Ang-(1-7) treatment as a possible strategy for pharmacological management of fibrosis-associated ED in aging. Strengths & LimitationsThe link between the RAS and penile fibrosis, indicated by a holistic screening of different ED markers, was confirmed by in vivo and in vitro data. However, results, presented in the manuscript, need to be further reinforced by human data. Important to note, the main goal of the study was to characterize RAS regulation in aging condition rather than state any causal relationships. ConclusionPresent study characterizes RAS regulation in aging-associated ED and indicates its important role in cavernosal fibrosis.Bragina ME, Costa-Fraga F, Sturny M, et al. Characterization of the Renin-Angiotensin System in Aged Cavernosal Tissue and its Role in Penile Fibrosis. J Sex Med 2020;17:2129–2140.

Re: Evaluating the Safety and Potential Activity of URO-902 (hMaxi-K) Gene Transfer by Intravesical Instillation or Direct Injection into the Bladder Wall in Female Participants with Idiopathic (Non-Neurogenic) Overactive Bladder Syndrome and Detrusor Overactivity from Two Double-Blind, Imbalanced, Placebo-Controlled Randomized Phase 1 Trials.

No AccessJournal of UrologyUrological Survey1 Oct 2020Re: Evaluating the Safety and Potential Activity of URO-902 (hMaxi-K) Gene Transfer by Intravesical Instillation or Direct Injection into the Bladder Wall in Female Participants with Idiopathic (Non-Neurogenic) Overactive Bladder Syndrome and Detrusor Overactivity from Two Double-Blind, Imbalanced, Placebo-Controlled Randomized Phase 1 Trials Alan J. WeinMD, PhD (hon), FACS Alan J. WeinAlan J. Wein More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000001223.02AboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail "Re: Evaluating the Safety and Potential Activity of URO-902 (hMaxi-K) Gene Transfer by Intravesical Instillation or Direct Injection into the Bladder Wall in Female Participants with Idiopathic (Non-Neurogenic) Overactive Bladder Syndrome and Detrusor Overactivity from Two Double-Blind, Imbalanced, Placebo-Controlled Randomized Phase 1 Trials." The Journal of Urology, 204(4), pp. 884–885 Suggested Reading : Diagnosis and treatment of overactive bladder (non‐neurogenic) in adults: AUA/SUFU Guideline amendment 2019. J Urol 2019; 202: 558. Link, Google Scholar : The successful long‐term treatment of age related erectile dysfunction with hSlo cDNA in rats in vivo. J Urol 2003; 170: 285. Link, Google Scholar © 2020 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 204Issue 4October 2020Page: 884-885 Advertisement Copyright & Permissions© 2020 by American Urological Association Education and Research, Inc.MetricsAuthor Information Alan J. Wein More articles by this author Expand All Advertisement Loading ...

The protective effect of Ferula elaeochytris on age-related erectile dysfunction

Ethnopharmacological relevanceFerula elaeochytris Korovin (FE) is a perennial medicinal plant of Apiaceae family. Ferula elaeochytris Korovin, known as ‘Çakşır’ in Anatolia, is widely used as an aphrodisiac as well as antioxidant, anti-inflammatory, and anti-diabetic.Aim of the study: Erectile dysfunction (ED) is a serious public health problem that has a high prevalence and negatively affects the quality of life in elderly men. In the treatment and prophylaxis of many diseases, because of widely increasing use of plant extracts as therapeutic agents, preclinical studies related to plant extracts are becoming more important by the day. In this study, we aimed to investigate the protective effect of Ferula elaeochytris Korovin (FE) root extract on age-related ED. Materials and methodsSeventy-two male Wistar albino rats were equally divided into four groups: 4-month aged rats (Y), 24-month aged rats (AG), and FE-administered (20 and 40 mg/kg/day; oral gavage; over 8 weeks) 24-month aged rats (AG + FE). The measurements included: changes in smooth muscle cells and collagen fibrils, tumor necrosis factor alpha (TNF-α), penile neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS) expression, serum testosterone concentrations (ST), neurogenic- and endothelial-dependent relaxations of the corpus cavernosum (CC), intracavernosal pressure/mean arterial pressure (ICP/MAP), area under the curve (total ICP), total antioxidant status (TAS), and total oxidant status (TOS) on corpus cavernosal tissue. ResultsThese results have an important role in the development of ED. ICP/MAP, total ICP, eNOS/nNOS expressions and ST levels increased in AG+40 mg FE group compared to the AG group, whereas TNF-α levels decreased and oxidative and antioxidant parameters balanced. ConclusionsOur findings show that FE may have a useful effect on decelerating the development of age-related ED.

Restoration of erectile function by suppression of corporal apoptosis and oxidative stress with losartan in aged rats with erectile dysfunction.

The prevalence of erectile dysfunction (ED) increases progressively with age, but its potential pathophysiology has not been fully demonstrated. Hence, this article was aimed to identify the functional and morphological characterization of the corpus cavernosum in aged rats and to evaluate the effects of the Angiotensin II type 1 receptor antagonist losartan on age-related ED (AED). A total of 40 young and aged Sprague Dawley rats were randomly divided into four groups (n=10 per group): young rats as normal controls (YNC) group; aged rats with normal erectile function (ANC) group; aged rats with ED (AED) group; and a losartan-treated AED (AED+Losartan) group. The treated group received losartan (30mg/kg) once daily oral gavage for 4weeks. Erectile function was measured by the ratio of peak intracavernous pressure (ICP)/mean arterial pressure (MAP), and relevant tissues were harvested for transmission electron microscopy, Immunohistochemistry, Masson's trichrome staining, TUNEL, caspase-3 activity assay and Western blot. The AED group exhibited decreases in erectile response and increases in the role of apoptosis, fibrosis as well as oxidative stress, compared with the control groups. After daily administration of losartan for four weeks, it could slightly restore erectile function and significantly attenuate corporal apoptosis, fibrosis, and oxidative stress of AED. However, the result was still not comparable with that of the control groups. Moreover, the expression levels of p-Bad/Bad and p-AKT/AKT were significantly lower, whereas the expression levels of Bax/Bcl-2, Nrf2/Keap-1, Fibronectin, HO-1, and caspase-3 activity were significantly higher in the AED groups and while losartan could significantly attenuate these changes of AED, it was still not comparable with that of the control groups. Our results indicated that administration of losartan not merely restored erectile function, but also significantly prevented corporal apoptosis and oxidative stress in AED by suppressing the Akt/Bad/Bax/caspase-3 and Nrf2/Keap-1 pathways.

169 COMP-4 Reduces Oxidative Stress Markers in the Rat Corpora Cavernosa and Media of the Penile Dorsal Artery

In age related erectile dysfunction, there is a progressive aging dependent reduction in the corporal smooth muscle cells (CSMCs) with replacement by collagen. To combat these histological alterations, the aging CSMCs begin to produce nitric oxide (NO) via upregulation of the normally quiescent inducible nitric oxide synthase (iNOS) enzyme. In a recent study, it was shown that COMP-4, a compound composed of ginger, muira puama, Paulina cupana, and L-citrulline, stimulated production of iNOS in vitro while treatment of middle aged rats for two months with oral COMP-4 resulted in an increase in the CSMC content with a resultant improvement in erectile function. In this study, we investigate whether activation of the iNOS-NO-cGMP pathway by COMP-4 can reduce oxidative stress in both the CSMC and the media of the extra-corporal penile dorsal artery (PDA). Ten-month old Sprague Dawley rats were treated daily for two months with either vehicle (n=6) or COMP-4 (n=6) prior to euthanization. Rat penises were excised and then processed for immunohistochemical staining of both the cavernosa and PDA. Apoptosis was determined by TUNEL assay and cell proliferation by PCNA staining. Markers of oxidative stress were determined by nitrotyrosine (NT) and myeloperoxidase (MPO) while anti-oxidative activity was determined with heme oxygenase 1 (HO-1) expression. Cellular oxidative stress within the cavernosa was measured using the redox glutathione ratio (GSH/GSSG) from CSMC culture treated with or without COMP-4.

MicroRNA-145 engineered bone marrow-derived mesenchymal stem cells alleviated erectile dysfunction in aged rats

BackgroundAging is one of the dominant factors contributing to erectile dysfunction (ED), and effective treatments for age-associated ED are urgently demanded. In this study, the therapeutic efficiency of bone marrow-derived mesenchymal stem cells (BMSCs) overexpressing microRNA-145 (miR-145) was evaluated in ED.MethodsSixty male Sprague-Dawley rats (24 months old) were randomly divided into 4 treatment groups (n = 15/group): PBS (control), BMSCs, BMSCs transfected with a blank vector (vector-BMSCs), and BMSCs transfected with a lentivirus overexpressing miR-145 (OE-miR-145-BMSCs). Fourteen days after transplantation of BMSCs, erectile function was evaluated by measuring intra-cavernous pressure (ICP) and mean arterial pressure (MAP). Subsequently, penile erectile tissues were harvested and subjected to Masson staining, qRT-PCR, immunofluorescence staining, dual luciferase assay, and Western blot analysis.ResultsFourteen days after transplantation, the ICP/MAP was 0.79 ± 0.05 in the OE-miR-145-BMSC group, 0.61 ± 0.06 in the BMSC group, 0.57 ± 0.06 in the vector-BMSC group, and 0.3 ± 0.01 in the PBS group. Treatment with OE-miR-145-BMSCs significantly improved ED (P < 0.05), and the treatment increased the smooth muscle content in the penis tissues of ED rats (P < 0.05). In the OE-miR-145-BMSC group, the expression levels of α-SMA, desmin, and SM-MHC were higher than they were in the other ED groups (P < 0.05). In addition, the levels of collagen 1, MMP2, and p-Smad2 in the BMSC-treated group, especially in the OE-miR-145-BMSC group, were lower than those in the control group (P < 0.05).ConclusionsMicroRNA-145 engineered BMSCs effectively attenuate age-related ED. Transplantation of miR-145-overexpressing BMSCs may provide a promising novel avenue for age-associated ED therapy.

Bupropion use in the first trimester of pregnancy

Cavernosal fibrosis is an important pathologic condition leading to erectile dysfunction (ED). The etiologies of cavernosal fibrosis include aging, diabetes mellitus, castration, cavernosal nerve injury during radical prostatectomy, hypertension, and Peyronie disease.To summarize published studies investigating suppression of cavernosal fibrosis in rat models of ED of various etiologies.A literature search was conducted using PubMed. Relevant studies were identified using search terms such as erectile dysfunction, penis, fibrosis, and rat models.We reviewed representative literature studies on the mechanisms and suppression of cavernosal fibrosis in rat models of ED.The underlying mechanisms and potential therapeutic strategies suggested thus far for cavernosal fibrosis in rat models of ED were as follows. For age-related ED involving oxidative stress and tumor growth factor-β1 (TGF-β1)–driven pathways such as RhoA-ROCK1-LIMK2-cofilin or p42-44 and mitogen-activated protein kinase, proposed therapeutic strategies included phosphodiesterase type 5 inhibitors (PDE5Is), kallikrein-kinin system stimulators, and calorie restriction. For diabetes-related ED involving angiotensin-II– and TGF-β1–driven Smad and non-Smad pathways, TGF-β1-Wnt10b, and histone deacetylase (HDAC)-TGF-β1 pathways, positive therapeutic results were obtained with PDE5Is, TGF-β1 antagonists, HDAC inhibitors, antioxidants, sphingosine-1-phosphate receptor modulators (fingolimod), angiotensin-II antagonists, stem cell therapy, and antidiabetic drugs. For cavernosal nerve injury-associated ED involving TGF-β1–driven pathways (Smad or RhoA-ROCK1-LIMK2-cofilin), Sonic hedgehog signaling, angiotensin-II–Smad, and HDAC4–TGF-β1–Smad signaling triggered by cavernosal hypoxia, PDE5Is, angiotensin-II antagonists, stem cell therapy, HDAC inhibitors, Sonic hedgehog administration, ROCK inhibitors, and LIMK2 inhibitors have shown positive results. For testosterone deficiency-associated ED, TGF-β1–driven pathways were found to be responsive to testosterone supplementation. For hypertensive ED, positive therapeutic results were obtained with angiotensin-II antagonists. For Peyronie disease involving TGF-β1 or myostatin signaling, proposed therapeutic strategies included intra-tunical injection of TGF-β receptor inhibitors or adipose tissue-derived stem cells and HDAC2 small hairpin RNA.Several signaling pathways appear to be responsible for the development of cavernosal fibrosis related to ED of various etiologies. Some therapeutic success has been achieved in animal models, but further research focusing on mechanism-specific targeted therapies is needed.Cho MC, Song WH, Paick J-S. Suppression of Cavernosal Fibrosis in a Rat Model. Sex Med Rev 2018;6:572–582.

Human tissue kallikrein 1 ameliorates erectile function via modulation of macroautophagy in aged transgenic rats.

Previously, we have demonstrated that human tissue kallikrein 1 (hKLK1) improves age-related erectile dysfunction (ED). Autophagy has been implicated in age-related diseases, including ED. However, the molecular mechanisms underlying hKLK1-mediated amelioration of age-related ED via regulation of autophagy remains unknown. To explore the potential mechanism, male wild-type Sprague-Dawley rats (WTR) and transgenic rats harboring human KLK1 (TGR) were bred till 4 or 18months of age and divided into three groups: young WTR (yWTR) as the control group, aged WTR (aWTR) group, and aged TGR (aTGR) group. The erectile function of each rat was evaluated using cavernous nerve electrostimulation. The ratio of intracavernous pressure/mean arterial pressure (ICP/MAP) and total ICP were also measured. Western blotting, immunohistochemistry, and transmission electron microscopy were performed to detect the levels of autophagy. The expression levels of related signaling pathways were determined by western blotting and immunohistochemistry. We found that hKLK1 improved the impaired erectile function of aged rats. Compared to the yWTR and aTGR groups, the aWTR group showed reduced smooth muscle/collagen ratio, fewer autophagosomes, and lower expression of Beclin 1 and LC3-II, which indicate impaired smooth muscle function and low level of autophagy in the smooth muscle cells. Moreover, the PI3K/Akt/mTOR signaling pathway, which is considered to be a negative regulator of autophagy, was upregulated in the aWTR group. hKLK1 may partially restore erectile function in aged transgenic rats by upregulating protective autophagy via the PI3K/Akt/mTOR pathway. These observations indicate that hKLK1 is a potential gene therapy candidate for age-related ED.

Suppression of Cavernosal Fibrosis in a Rat Model.

Cavernosal fibrosis is an important pathologic condition leading to erectile dysfunction (ED). The etiologies of cavernosal fibrosis include aging, diabetes mellitus, castration, cavernosal nerve injury during radical prostatectomy, hypertension, and Peyronie disease. To summarize published studies investigating suppression of cavernosal fibrosis in rat models of ED of various etiologies. A literature search was conducted using PubMed. Relevant studies were identified using search terms such as erectile dysfunction, penis, fibrosis, and rat models. We reviewed representative literature studies on the mechanisms and suppression of cavernosal fibrosis in rat models of ED. The underlying mechanisms and potential therapeutic strategies suggested thus far for cavernosal fibrosis in rat models of ED were as follows. For age-related ED involving oxidative stress and tumor growth factor-β1 (TGF-β1)-driven pathways such as RhoA-ROCK1-LIMK2-cofilin or p42-44 and mitogen-activated protein kinase, proposed therapeutic strategies included phosphodiesterase type 5 inhibitors (PDE5Is), kallikrein-kinin system stimulators, and calorie restriction. For diabetes-related ED involving angiotensin-II- and TGF-β1-driven Smad and non-Smad pathways, TGF-β1-Wnt10b, and histone deacetylase (HDAC)-TGF-β1 pathways, positive therapeutic results were obtained with PDE5Is, TGF-β1 antagonists, HDAC inhibitors, antioxidants, sphingosine-1-phosphate receptor modulators (fingolimod), angiotensin-II antagonists, stem cell therapy, and antidiabetic drugs. For cavernosal nerve injury-associated ED involving TGF-β1-driven pathways (Smad or RhoA-ROCK1-LIMK2-cofilin), Sonic hedgehog signaling, angiotensin-II-Smad, and HDAC4-TGF-β1-Smad signaling triggered by cavernosal hypoxia, PDE5Is, angiotensin-II antagonists, stem cell therapy, HDAC inhibitors, Sonic hedgehog administration, ROCK inhibitors, and LIMK2 inhibitors have shown positive results. For testosterone deficiency-associated ED, TGF-β1-driven pathways were found to be responsive to testosterone supplementation. For hypertensive ED, positive therapeutic results were obtained with angiotensin-II antagonists. For Peyronie disease involving TGF-β1 or myostatin signaling, proposed therapeutic strategies included intra-tunical injection of TGF-β receptor inhibitors or adipose tissue-derived stem cells and HDAC2 small hairpin RNA. Several signaling pathways appear to be responsible for the development of cavernosal fibrosisrelated to ED of various etiologies. Some therapeutic success has been achieved in animal models, but further research focusing on mechanism-specific targeted therapies is needed. Cho MC, Song WH, Paick J-S. Suppression of Cavernosal Fibrosis in a Rat Model. Sex Med Rev 2018;6:572-582.

Effect of ginger, Paullinia cupana, muira puama and l- citrulline, singly or in combination, on modulation of the inducible nitric oxide- NO-cGMP pathway in rat penile smooth muscle cells

IntroductionCOMP-4 is a natural compound-based dietary supplement consisting of the combination of ginger, Paullinia cupana, muira puama and l-citrulline, which when given long-term has been shown in the aged rat to a) upregulate iNOS in the penile smooth muscle cells (SMC), b) reverse the corporal SMC apoptosis and fibrosis associated with corporal veno-occlusive dysfunction (CVOD), and c) improve resulting erectile function. To elucidate the mechanism of how COMP-4 and its individual components modulate the iNOS-cGMP pathway, an in vitro study was conducted using a rat corporal primary SMC culture to determine its effect on NOS, soluble guanylate cyclase (sGC), cGMP and the phosphodiesterase 5 enzyme (PDE5). Materials and methodsPrimary SMC cultures using the explant technique were initiated by cutting small pieces of corporal tissue from 8 week old Sprague-Dawley rats. The SMC were grown in Dulbecco media with 20% fetal calf serum. The SMC were then incubated with or without COMP-4 (0.69 mg/ml) or its ingredients alone (ginger: 0.225 mg/ml; muira puama, Paullinia cupana and l-citrulline each at 0.9 mg/ml) for up to 24 h mRNA and protein were extracted and used for the determination of NOS, sGC and PDE5 content. cGMP content was determined by ELISA. L-NIL (4 μM) was used as an inhibitor of iNOS activity. ResultsCompared to the control values, COMP-4 upregulated expression of cGMP by 85%, induced a 42 fold increase in sGC as well as a 15 fold increase in both iNOS protein and mRNA content while it decreased both PDE5 mRNA and protein content each by about 50%. L-NIL completely inhibited the effect of COMP-4 on cGMP production. When compared with each of the individual four components of COMP-4, it appears that COMP-4 itself had the most profound effect in modulating each one the specific steps within the iNOS-cGMP pathway. ConclusionsThis in vitro study demonstrates that COMP-4 is capable of activating the endogenous cellular iNOS-cGMP pathway within the CSM cells, which is theorized to be responsible for reducing the fibrosis and apoptosis as well as the CVOD observed in the aging rat penis. Further studies will be necessary in order to determine whether supplementation of COMP-4 on a daily basis may be beneficial in halting or reversing this aging related erectile dysfunction in the clinical setting.

AB089. Impaired adenosine signaling influences erectile function in aging rats

BackgroundAs one of the most common disorders in old adult, erectile dysfunction (ED) remains attracting andrological physicians’ attention. The aim of this study is to investigate the alterations of adenosine signaling in the penis of aging rats, and the influence to erectile function.MethodsAccording to apomorphine test, the aging rats (18 months) with ED were selected as age-related erectile dysfunction (A-ED) group, and the young rats (2 months) were selected as normal control (NC) group. The intracavernosal pressure (ICP) measurements were conducted to evaluate the penile erectile function. Quantitative real-time polymerase chain reaction (RT-PCR) and Western Blot were used to detect the expression levels of genes and protein related to adenosine signaling in penis.ResultsCompared to NC group, the outcomes of ICP showed a decreasing trend in A-ED group. Expression of adenosine A2B receptor, adenosine deaminase (ADA), and phosphodiesterase type (5PDE5) were increased in A-ED group, and AMP deaminase type 1 (AMPD1) and 2 (AMPD2) were decreased in A-ED group. The results of Western Blot also showed an increasing trend of A2B receptor in A-ED group.ConclusionsRats with erectile dysfunction showed an impaired adenosine signaling, our study may provide a new sight for further study to improve the erectile function of A-ED patients.

MP81-14 HUMAN TISSUE KALLIKREIN 1 AMELIORATES ERECTILE FUNCTION VIA MODULATING AUTOPHAGY AND ACTIVATING HIF-1?/COX-2 PATHWAY IN AGED TRANSGENIC RATS

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology II1 Apr 2017MP81-14 HUMAN TISSUE KALLIKREIN 1 AMELIORATES ERECTILE FUNCTION VIA MODULATING AUTOPHAGY AND ACTIVATING HIF-1?/COX-2 PATHWAY IN AGED TRANSGENIC RATS Zhe Tang, Kai Cui, Yang Luan, Yajun Ruan, Tao Wang, Jun Yang, Shaogang Wang, and Jihong Liu Zhe TangZhe Tang More articles by this author , Kai CuiKai Cui More articles by this author , Yang LuanYang Luan More articles by this author , Yajun RuanYajun Ruan More articles by this author , Tao WangTao Wang More articles by this author , Jun YangJun Yang More articles by this author , Shaogang WangShaogang Wang More articles by this author , and Jihong LiuJihong Liu More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.2540AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Our previous studies have demonstrated that Human Tissue Kallikrein 1 (hKLK1) improved erectile function via several signaling pathways related to such as oxidative stress or corporal cavernosal fibrosis. However, the potential molecular mechanisms of hKLK1 inhibited age-related erectile dysfunction via modulating autophagy remains unknown. The aim of this article is to partly clarify the mechanisms of hKLK1 improving the erectile function in aged rats. METHODS Male wild-type Sprague-Dawley rats (WTR) and transgenic rats expressing the hKLK1 gene (TGR) were fed to 4 and 18 months of age, respectively, and divided into three groups: young WTR (yWTR) as the control, aged WTR (aWTR) and aged TGR (aTGR). Cavernous nerve electrostimulation was used to evaluate the erectile function of all rats. Transmission electron microscopy, immunohistochemistry, and western blotting were performed to determine the levels of autophagy. Related signaling pathways were detected by western blot and immunohistochemistry. RESULTS Compared with the yWTR group and aTGR group, the aWTR group showed (1)lower erectile function: lower intracavernosal pressure(ICP)/mean arterial pressure(MAP) ratio; (2) reduced expressions of eNOS and nNOS, lower NO level; (3) inhibited autophagy: decreased autophagosomes, lower expressions of BECN1 and LC3-II; and (4) low expression levels of PI3K/Akt/mTOR and Hif-1a/Cox-2 pathways. CONCLUSIONS The hKLK1 gene played a potential role of restoring erectile function in aged transgenic rats through modulating autophagy via PI3K/Akt/mTOR and Hif-1a/Cox-2 pathways. This finding provided evidence for hKLK1 being gene therapy method of age-related erectile dysfunction. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e1094 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Zhe Tang More articles by this author Kai Cui More articles by this author Yang Luan More articles by this author Yajun Ruan More articles by this author Tao Wang More articles by this author Jun Yang More articles by this author Shaogang Wang More articles by this author Jihong Liu More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

HP-01-003 Functional determinants of aging-related erectile dysfunction in rats

HP-01-003 Functional determinants of aging-related erectile dysfunction in rats

MP45-03 AGE RELATED PENILE HEMODYNAMIC IMPAIRMENT AND FIBROSIS: POSSIBLE ROLE OF GIV-WNT SIGNALING PATHWAYS

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology I1 Apr 2017MP45-03 AGE RELATED PENILE HEMODYNAMIC IMPAIRMENT AND FIBROSIS: POSSIBLE ROLE OF GIV-WNT SIGNALING PATHWAYS Tung-Chin (Mike) Hsieh, Sadhna Kanoo, Jay Parikh, Valmik Bhargava, and M. Raj Rajasekaran Tung-Chin (Mike) HsiehTung-Chin (Mike) Hsieh More articles by this author , Sadhna KanooSadhna Kanoo More articles by this author , Jay ParikhJay Parikh More articles by this author , Valmik BhargavaValmik Bhargava More articles by this author , and M. Raj RajasekaranM. Raj Rajasekaran More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2017.02.1421AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Pathophysiology/ molecular mechanisms of age-related penile hemodynamic impairment are unclear. Previous studies suggest age-related increase in tissue fibrosis in several organs including male genital tissues leading to impaired hemodynamics and male erectile dysfunction (ED). We tested the hypothesis that increased penile/ perineal (ischiocavernosus; ICM) muscle fibrosis during advanced aging is mediated by a novel fibrogenic cascade involving GIV/girdin and Wnt signaling pathways. A clear understanding of these molecular mechanisms involved in penile fibrosis would enable development of optimal strategies to treat ED. METHODS Young (n=3; 9 months) and old (n=6; 36 months) New Zealand White rabbits were anesthetized and penile tissue microcirculatory blood perfusion (perfusion units- PU) was measured using a novel PeriCam PSI system. Blood perfusion measurements were made before and after an intracavernosal injection of vasoactive agent- PGE1 (50 ng). Penile (corpus cavernosum) and ICM tissues were harvested and paraffin sections of these tissues were subjected to: (1) trichrome staining (marker of fibrosis) followed by image analysis for evaluation of fibrosis and (2) immunostaining for specific markers of fibrosis: 1. [beta]-catenin (central mediator of Wnt pathway) and 2. GIV/girdin. RESULTS Representative tracings of time-course of penile blood perfusion measurements in response to intracavernosal PGE1 as well as age-related penile perfusion changes in young and old rabbits are shown in figure A. Photomicrographs of showing trichrome as well as immunostaining for fibrosis markers (GIV and [beta]-catenin) are depicted in Fig B. A significant impairment in penile perfusion (Fig A) and increased fibrosis of penile/ICM tissues was observed in old rabbits (Fig B) when compared to the young animals. Immunostaining studies revealed positive labelling for both GIV and and [beta]-catenin and increased immunoreactivity in the tissues from old rabbits (Fig B). CONCLUSIONS Our findings suggest age-related increase in penile/ perineal muscle (ICM) fibrosis may contribute to increased incidence of ED in older men. Targeting GIV-Wnt pathways may be beneficial in preventing age-related ED for the aging population. © 2017FiguresReferencesRelatedDetails Volume 197Issue 4SApril 2017Page: e612 Advertisement Copyright & Permissions© 2017MetricsAuthor Information Tung-Chin (Mike) Hsieh More articles by this author Sadhna Kanoo More articles by this author Jay Parikh More articles by this author Valmik Bhargava More articles by this author M. Raj Rajasekaran More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...