Rat Model Of Erectile Dysfunction Research Articles

Dual Strategy with Adipose-Derived Stem Cells and l-arginine Recovered Cavernosal Functions in a Rat Model of Radical Prostatectomy.

As standard therapy for prostate cancer, radical prostatectomy causes cavernous nerve (CN) injury and increases fibrosis and hypoxia-induced penile structural alterations. This study aimed to determine the potential beneficial effects of adipose-derived stem cells (ADSCs) and l-arginine alone or in combination on the penile erection in a rat model of erectile dysfunction caused by bilateral cavernous nerve transection (CNT). Male rats (n = 35) were randomized into five groups: Sham-operated; CNT (4-weeks); CNT plus ADSCs (1 × 106 cells by intracavernosal injection); CNT plus l-arginine (4 weeks, 10 mg/kg/day, oral); and ADSCs combined with l-arginine in CNT. In vivo erectile responses and in vitro relaxant responses were measured. Western blot and immunohistochemistry analyses were used to determine the expression and localization of endothelial nitric oxide synthase, neuronal nitric oxide synthase, transforming growth factor-beta 1, hypoxia-inducible factor-1 alpha (HIF-1α), and apoptosis markers (Bax and Bcl-2). The ratio of smooth muscle to collagen and nerve regeneration were calculated using Masson's trichrome and nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase staining. The combined treatment restored diminished erectile responses, endothelium-dependent acetylcholine, and electrical field stimulation-induced relaxation of the corpus cavernosum in rats with CNT, whereas either monotherapy produced only partial improvements. All treatment regimens restored increases in the protein expression of HIF-1 and Bax in rats with CNT. The decrease in smooth muscle mass and NADPH-diaphorase-positive nerve fibers was partially ameliorated by monotherapy, whereas combined therapy led to recovery. These findings indicate that combined treatment with ADSCs and l-arginine may restore erectile function in rats with CNT by inhibiting hypoxia-induced neurotoxicity and preserving endothelium function and smooth muscle content.

(093) The Light-Controlled Nitric Oxide Donor “NORD-1” may Improve Erectile Function Without Hypotension and Tissue Damage in Rats With Neurogenic Erectile Dysfunction

Abstract Introduction Radical prostatectomy (RP) often causes erectile dysfunction (ED) because of cavernous nerve damage during surgery. This damage results in a decrease of nitric oxide (NO) in the penile corpus cavernosum. Therefore, NO supplementation is considered to be effective for post RP ED. However, NO has systemic side effects such as headache and hypotension. Recently, a red-light-controlled NO donor “NORD-1”, developed by our group, enables spatially and temporally controlled NO release by light irradiation. Our previous study reported that NORD-1 combined with red-light irradiation is effective for neurogenic ED model rats with bilateral cavernous nerve injury (BCNI). However, the side effects of the treatment have not yet been discussed. Objective The study aimed to investigate the potency of side effects associated with the combination of NORD-1 and red-light irradiation, using a ED rat model with BCNI. Methods In this study, eight-week-old male Wistar-ST rats were used. The rats were divided into four groups: Sham+SiR650 (control compound), Sham+NORD-1, BCNI+SiR650, and BCNI+NORD-1 (n=6). The sham or BCNI surgery was performed under anesthesia in each group. BCNI surgery was performed by crushing the cavernous nerves bilaterally. Four weeks post-surgery, erectile function before and after the injection of each reagent into the penile corpus cavernosum was evaluated based on the changes in intracavernous pressure (ICP) under electrostimulation of the cavernous nerve. After the administration of each reagent, the changes in ICP with and without light irradiation were also evaluated. ICP during electrostimulation was totally measured three times; once without reagent and light, once with reagent and without light and once with reagent and light. The variations in systemic arterial pressure during electrostimulation were also analyzed. After the experiment, penile samples were collected to observe NORD-1 localization using a fluorescence microscope. In addition, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay was performed to evaluate apoptosis in the corpus cavernousm. Results Before administering each reagent without light irradiation, the ICP/mean arterial pressure (MAP) ratios under stimulation in the BCNI+SiR650 and BCNI+NORD-1 groups were significantly lower than those in the Sham+SiR650 (P < 0.01) and Sham+NORD-1 (P < 0.05) groups. After administering each reagent without light irradiation, the ICP/MAP ratios in the BCNI+SiR650 and BCNI+NORD-1 groups remained significantly lower than those in the Sham+SiR650 and Sham+NORD-1 group (P < 0.05 and P < 0.01, respectively). After administering each reagent with light irradiation, the ICP/MAP ratios in the Sham+NORD-1 and BCNI+NORD-1 groups were significantly higher than those in the Sham+SiR650 (P < 0.01) and BCNI+NORD-1 (P < 0.01) groups, respectively. The changes of MAP during electrostimulation were not different under three different conditions in all four groups. Fluorescence of NORD-1 was detected only in corpus cavernousum which means it was not detected in urethra and dorsal vein of penis. Apoptosis was not detected in corpus cavernousum in all four groups. Conclusions The combination of NORD-1 and light irradiation is effective for neurogenic ED in rats with cavernous nerve injury. The treatment may not cause hypotension and tissue damage. Disclosure No

(097) A Novel Therapeutic Strategy for Recovery from Cavernous Veno-Occlusive Dysfunction through Chronic Administration of Hepatocyte Growth Factor and a JNK Inhibitor in a Rat Model of Cavernous Nerve Injury

Abstract Introduction Structural alterations of penis including cavernosal apoptosis and fibrosis induce venous leakage in the corpus cavernosum or cavernosal veno-occlusive dysfunction, a key pathophysiology of erectile dysfunction (ED) after radical prostatectomy. Objective We aimed to determine if JNK inhibition in combination with intracavernosal administration of hepatocyte growth factor (HGF) would completely restore cavernosal veno-occlusive function (CVOF) in a rat model of ED induced by bilateral CN crush injury (CNCI). Methods A total of forty-two male Sprague-Dawley rats were randomly assigned to sham control (S), CNCI (I) and CNCI treated with combination of JNK inhibitors and HGF (J+H) for 5 weeks after surgery. All rats in each group were evaluated by dynamic infusion cavernosometry (DIC), caspase-3 activity assay, Masson’s trichrome staining, immunohistochemical staining of α-smooth muscle actin (α-SMA) and immunoblotting at 5 weeks after surgery. Results Regarding CVOF, Group I showed a decreased papaverine response, an increased maintenance and drop rates compared to Group S. Group J+H showed significant improvements in the three DIC parameters compared to Group I. There were no differences in papaverine response and drop rate between Group J+H and Group S. Regarding the structural integrity of corpus cavernosum, Group I had an increased caspase-3 activity, decreased SM/collagen ratio, decreased content of SM, decreased protein expression of PECAM-1, and decreased phosphorylation of c-Jun and c-Met. Group J+H had significant alleviation in the histological and molecular derangement compared to Group I. There were no differences in caspase-3 activity, SM content, PECAM-1 protein expression, c-Jun phosphorylation and c-Met phosphorylation between Group J+H and Group S. Conclusions Our data indicate that chronic administration of JNK inhibitors and HGF may restore CVOF to the level comparable to sham control level through preservation of structural integrity of the corpus cavernosum. Combination of JNK inhibition and HGF might be a potential therapeutic option for recovering from CVOD. Disclosure No

Chronic treatment with a combination of hepatocyte growth factor and JNK inhibitor ameliorates cavernosal veno-occlusive dysfunction: a rat model of cavernous nerve injury.

Structural alterations of the penis, including cavernosal apoptosis and fibrosis, induce venous leakage into the corpus cavernosum or cavernosal veno-occlusive dysfunction, a key pathophysiology associated with erectile dysfunction after radical prostatectomy. We hypothesized that the effect of JNK inhibitors on reducing apoptosis and hepatocyte growth factor (HGF) on inducing tissue regeneration could be another treatment mechanism of erectile dysfunction after radical prostatectomy. To investigate whether JNK inhibition combined with intracavernosal administration of HGF can completely preserve cavernosal veno-occlusive function (CVOF) in a rat model of erectile dysfunction induced via bilateral cavernosal nerve crush injury (CNCI). A total of 42 male Sprague-Dawley rats were randomly assigned to sham control (group S), CNCI (group I), and CNCI treated with a combination of JNK inhibitor and HGF (group J + H) for 5weeks after surgery. Rats in each group were evaluated via dynamic infusion cavernosometry (DIC), caspase-3 activity assay, Masson trichrome staining, immunohistochemical staining of α-smooth muscle actin, and immunoblotting at 5weeks after surgery. Regarding CVOF, group I showed decreased papaverine response, increased maintenance, and drop rates of DIC when compared with group S. Group J + H showed significant improvement in the 3 DIC parameters vs group I. No differences in the 3 DIC parameters were found between group J + H and group S. Regarding the structural integrity of the corpus cavernosum, group I showed increased caspase-3 activity, decreased smooth muscle (SM):collagen ratio, decreased SM content, decreased protein expression of PECAM-1, and decreased phosphorylation of c-Jun and c-Met. Group J + H showed significant attenuation in histologic and molecular derangement as compared with group I. There were no differences in caspase-3 activity, SM content, SM:collagen ratio, PECAM-1 protein expression, c-Jun phosphorylation, and c-Met phosphorylation between groups J + H and S. Our results suggest that antiapoptotic and regenerative therapy for the corpus cavernosum is a potential mechanism of penile rehabilitation after radical prostatectomy. This study provides evidence that combination treatment of JNK inhibitor and HGF preserves erectile function by restoring corporal SM and endothelium. However, additional human studies are needed to confirm the clinical effect. Chronic treatment with JNK inhibitor and HGF may preserve CVOF to levels comparable to sham control by preserving the structural integrity of the corpus cavernosum and so represents a potential therapeutic option for preventing the development of cavernosal veno-occlusive dysfunction.

Intracavernous injection of platelet-rich plasma reverses erectile dysfunction of chronic cavernous nerve degeneration through reduction of prostate hyperplasia evidence from an aging-induced erectile dysfunction rat model.

Age-induced erectile dysfunction (ED) is a convoluted medical condition, and restoring erectile function (EF) under geriatric conditions is highly complicated. Platelet-rich plasma (PRP) treatment is an inexpensive cell-based therapeutic strategy. We have aimed to restore EF in aged-ED rats with PRP as a therapeutic tool. Male rats were grouped into aged and young according to age. The young rats were considered as normal control (NC) and treated with saline. Aged were further divided into 2 groups and treated with intracavernous (IC) PRP and saline. Treatment was scheduled at the 9th and 10th week for NC and 41th and 42th week for aged-ED rats, with EF analysis scheduled on the 12th week for NC and 44th week for aged-ED rats, respectively. Erectile response, immunofluorescence staining, and electron microscopic analyses were performed. IC PRP treatment effectively reduced prostate hyperplasia(PH). EF response indicated a significant increase in crucial EF parameters in PRP-treated aged-ED rats. Histological evidence denoted a rigid and restored development of tunica adventitia of the dorsal artery, decreased vacuolation of the dorsal penile nerve, and structural expansion of the epineurium. Masson's trichrome and immunostaining results affirmed an elevated expression of α-smooth muscle actin(α-SMA) in the corpus cavernosum(CC). Ultrastructure findings revealed that PRP effectively rejuvenated degenerating nerves, preserved endothelium and adherent junctions of corporal smooth muscle, and restored the axonal scaffolds by upregulating neurofilament-H (NF-H)expression. Finally, PRP enhanced neural stability by enhancing the axonal remyelination processes in aged-ED rats. Hence, PRP treatment was proven to restore EF in aged-ED rats, which was considered a safe, novel, cost-effective, and hassle-free strategy for EF restoration in geriatric patients.

Overexpression of PRDX2 in Adipose-Derived Mesenchymal Stem Cells Enhances the Therapeutic Effect in a Neurogenic Erectile Dysfunction Rat Model by Inhibiting Ferroptosis.

Neurogenic erectile dysfunction (NED) is a common and serious complication after pelvic surgery. The clinical translation of adipose-derived mesenchymal stem cell (ADSC) therapies in NED remains a major challenge due to their low survival rate and limited therapeutic effect. Peroxiredoxin 2 (PRDX2) is a member of the peroxidase family that exerts its therapeutic effects by inhibiting oxidative stress (OS) and ferroptosis, and PRDX2 is expected to enhance the therapeutic effect of ADSCs in treating NED. The purpose of this study was to investigate whether PRDX2 could improve the survival of ADSCs and determine whether overexpression of PRDX2 in ADSCs (PRDX2-ADSCs) could enhance the therapeutic effect of NED. This study investigated the potential role of PRDX2-ADSCs through a NED model induced by bilateral cavernous nerve injury (BCNI) and three in vitro models established by H2O2-stimulated ADSCs, H2O2-stimulated corpus cavernosum smooth muscle cells (CCSMCs), and RSL3-stimulated CCSMCs. We found that PRDX2 could significantly improve the viability of ADSCs by suppressing apoptosis and OS in H2O2-stimulated ADSCs. We also found that BCNI triggered ferroptosis of the corpus cavernosum, which was manifested by increased reactive oxygen species (ROS), total iron content, and MDA as well as decreased SOD and GSH. Our results further demonstrated changes in the expression of key proteins (GPX4 and ACSL4) in the ferroptosis pathway, whereas PRDX2-ADSCs ameliorated BCNI-induced erectile dysfunction and ferroptosis of the corpus cavernosum in NED rats. Consistently, PRDX2-ADSCs attenuated OS in H2O2-stimulated CCSMCs and inhibited ferroptosis in RSL3-stimulated CCSMCs, as evidenced by the decrease in ROS, total iron content, and MDA and the increase in SOD and GSH together with changes in ferroptosis-related protein (GPX4 and ACSL4) expression. In conclusion, overexpression of PRDX2 in ADSCs enhanced the therapeutic effect in a rat model of neurogenic erectile dysfunction by inhibiting ferroptosis via regulation of the GPX4/ACSL4 axis.

Pathway analysis of microarray data from corpora cavernosal tissue of patients with a prostatectomy or Peyronie disease in comparison with a cavernous nerve-injured rat model of erectile dysfunction.

Patients with a prostatectomy are at high risk of developing erectile dysfunction (ED) that is refractory to phosphodiesterase type 5 inhibitors. The cavernous nerve (CN) is frequently damaged during prostatectomy, causing loss of innervation to the penis. This initiates corpora cavernosal remodeling (apoptosis and fibrosis) and results in ED. To aid in the development of novel ED therapies, the current aim was to obtain a global understanding of how signaling mechanisms alter in the corpora cavernosa with loss of CN innervation that results in ED. Microarray and pathway analysis were performed on the corpora cavernosal tissue of patients with a prostatectomy (n = 3) or Peyronie disease (control, n = 3). Results were compared with an analysis of a Sprague-Dawley rat CN injury model (n = 10). RNA was extracted by TRIzol, DNase treated, and purified by a Qiagen Mini Kit. Microarray was performed with the Human Gene 2.0 ST Array and the RU34 rat array. Differentially expressed genes were identified through several analytic tools (ShinyGO, Ingenuity, WebGestalt) and databases (GO, Reactome). A 2-fold change was used as the threshold for differential expression. Pathways that were altered (up- or downregulated) in response to CN injury in the prostatectomy patients and a rat CN injury model were determined. Microarray identified 197 differentially expressed protein-coding genes in the corpora cavernosa from the prostatectomy cohort, with 100 genes upregulated and 97 genes downregulated. Altered signaling pathways that were identified that affect tissue morphology included the following: neurologic disease, cell death and survival, tissue and cellular development, skeletal and muscle development and disorders, connective tissue development and function, tissue morphology, embryonic development, growth and proliferation, cell-to-cell signaling, and cell function and maintenance. These human pathways have high similarity to those observed in the CN-injured rat ED model. Significant penile remodeling continues in patients long after the acute surgical injury to the CN takes place, offering the opportunity for clinical intervention to reverse penile remodeling and improve erectile function. Understanding how signaling pathways change in response to CN injury and how these changes translate to altered morphology of the corpora cavernosa and ensuing ED is critical to identify strategic targets for therapy development. Altered signaling in pathways that regulate tissue homeostasis, morphogenesis, and development was identified in penes of patients with a prostatectomy, and competitive forces of apoptosis and proliferation/regeneration were found to compete to establish dominance after CN injury. How these pathways interact to regulate penis tissue homeostasis is a complex process that requires further investigation.

A3 adenosine receptor allosteric modulator CF602 reverses erectile dysfunction in a diabetic rat model.

Adenosine plays a major role in erection by binding to its receptors and activating pathways resulting in increased arterial blood flow and intracavernosal pressure (ICP). CF602, an allosteric modulator of the A3 adenosine receptor (A3AR), increases the binding affinity of the endogenous adenosine to the receptor. We examined the effect of CF602 on resolving erectile dysfunction (ED) in a diabetic ED rat model (streptozotocin-induced diabetic rats that were screened for ED using the apomorphine test). ED was assessed by measuring ICP and main arterial pressure (MAP) during electrostimulation of the cavernosal nerve. A single dose of CF602 or placebo was applied either topically (100 μl from a 100 nM or 500 nM solution) or orally (100, 200 or 500 μg/kg) prior to erectile function assessment. A significant dose-dependent improvement in the ICP:MAP ratio without a change in MAP was recorded with the topical and oral CF602 treatments. A significant increase in smooth muscle:collagen ratio, vascular endothelial growth factor and endothelial nitric oxide synthase was also observed in both administration modes. In conclusion, topical and oral treatment with CF602 significantly improved erectile function, supporting its further evaluation as a treatment for ED.

MP38-03 COMBINATION TREATMENT WITH A JNK INHIBITOR AND HEPATOCYTE GROWTH FACTOR FOR RESTORATION OF ERECTILE FUNCTION IN A RAT MODEL OF CAVERNOSAL NERVE INJURY: COMPARISON WITH A JNK INHIBITOR ALONE OR HEPATOCYTE GROWTH FACTOR ALONE

You have accessJournal of UrologyCME1 May 2022MP38-03 COMBINATION TREATMENT WITH A JNK INHIBITOR AND HEPATOCYTE GROWTH FACTOR FOR RESTORATION OF ERECTILE FUNCTION IN A RAT MODEL OF CAVERNOSAL NERVE INJURY: COMPARISON WITH A JNK INHIBITOR ALONE OR HEPATOCYTE GROWTH FACTOR ALONE Junghoon Lee, Sangjun Yoo, Min Soo Choo, Hwancheol Son, Hyeon Jeong, Soo Woong Kim, and Min Chul Cho Junghoon LeeJunghoon Lee More articles by this author , Sangjun YooSangjun Yoo More articles by this author , Min Soo ChooMin Soo Choo More articles by this author , Hwancheol SonHwancheol Son More articles by this author , Hyeon JeongHyeon Jeong More articles by this author , Soo Woong KimSoo Woong Kim More articles by this author , and Min Chul ChoMin Chul Cho More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002592.03AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: The structural derangements of the corpus cavernosum are known to be potential causes of intractable impotence or to limit efficacy of PDE5 inhibitors by inducing cavernosal venoocclusive dysfunction, a key pathophysiology of postprostatectomy erectile dysfunction. The aim of this study was to determine if combined administration of a JNK inhibitor and HGF (hepatocyte growth factor) would restore erectile function through both antiapoptotic and regenerative effects in a rat model of erectile dysfunction induced by cavernous nerve crush injury (CNCI), and to compare it with administration of a JNK inhibitor alone or HGF alone. METHODS: We randomized 70 rats into five experimental groups for two weeks from the day of CNCI: a sham surgery group (S), a CNCI (I) group, a group treated with once-daily intraperitoneal administration of 10.0 mg/kg of JNK inhibitor (J), a twice-weekly intracavernosal administration of 4.2 μg HGF group (H), and a combined treatment with 10.0 mg/kg JNK inhibitor and 4.2 μg HGF group (J+H). We investigated erectile responses to electrostimulation, histological staining, caspase-3 activity assay, and immunoblotting at two weeks following surgery. RESULTS: The three treatment groups showed improvements in the erectile responses (ICP/MAP or AUC/MAP ratio) after stimulation at all voltages as compared to the I group. The erectile responses in the J+H group were greater than those in the J or H group. The erectile responses in the J+H group were not generally different from those in the S-group. Regarding the cavernosal apoptosis, the caspase-3 activity and c-Jun phosphorylation in the J and J+H groups were improved as compared to the I group, whereas the caspase-3 activity in the H group partially improved. Regarding the endothelial status and smooth muscle (SM) content, the protein expression of PECAM-1, eNOS phosphorylation, and α-SM actin content in the J+H group were normalized, although those in the J or H group were partially restored. Phosphorylation of cMet was normalized in the H and J+H groups. CONCLUSIONS: Our data indicates that combination therapy with a JNK inhibitor and HGF can generally normalize erectile function through antiapoptosis and the preservation of cavernosal endothelium or SM in a rat CN injury model. The combined treatment with these two kinds of agents also appears to be superior to the respective agent alone in terms of the therapeutic effects. Source of Funding: This research was supported by the National Research Foundation of Korea, grant 2018R1D1A1A09083833, and by the JEIL Pharmaceutical Co., LTD, grant 01-2018-33 © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e619 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Junghoon Lee More articles by this author Sangjun Yoo More articles by this author Min Soo Choo More articles by this author Hwancheol Son More articles by this author Hyeon Jeong More articles by this author Soo Woong Kim More articles by this author Min Chul Cho More articles by this author Expand All Advertisement PDF DownloadLoading ...

Treatment with levothyroxine and testosterone ameliorated erectile dysfunction in rat model of propylthiouracil-induced hypothyroidism

ABSTRACT Introduction The endocrine disease frequently interrupts sexual function, and sexual dysfunction may signal serious endocrine disease. Recent studies have claimed an association with hypothyroidism, low testosterone (T) levels and erectile dysfunction (ED), but causality is unclear. There is no study on the experimental model of hypothyroidism that was induced in rats by administration of propylthiouracil (PTU). Objective The purpose of this study was to investigate the deleterious effects of PTU-induced hypothyroidism on ED and T levels. In addition, Levothyroxine is known worldwide as the most common and least effective drug for controlling hypothyroidism. Methods Adult Sprague-Dawley rats (n = 35) were divided into five groups; control, PTU-induced hypothyroidism, PTU+ Levothyroxine, PTU+ Sustanon (a mixture of 4 types of T: propionate, phenylpropionate, isocaproate, decanoate) and PTU+ Levothyroxine + Sustanon. PTU was given in drinking water (0.05%) for 6 weeks. 4 weeks after the beginning of PTU treatment, Levothyroxine (20 mg/kg/day, oral) and Sustanon (10 mg/kg, I.M., once/week) were given for 2 weeks. Serum levels of total T, triiodothyronine (T3) and thyroxine (T4) were determined. The ratio of intracavernosal pressure (ICP) to mean arterial pressure (MAP) and total ICP were measured by cavernous nerve stimulation. The relaxant and contractile responses were evaluated on in vitro preparations of corpus cavernosum (CC) strips. The expression and localization of neuronal (nNOS), endothelial NOS (eNOS) and phosphodiesterase type 5 (PDE5) were determined using Western blotting and immunohistochemistry. The relative area of smooth muscle to collagen was measured using Masson trichrome staining. Results The rat model of hypothyroidism showed a significant decline in serum levels of total T, T3 and T4. Levothyroxine increased T3 and T4 levels while Sustanon normalized only total T levels. In addition, combined treatment of Levothyroxine and Sustanon enhanced all hormone levels. Rats with hypothyroidism displayed the lowest ICP/MAP ratio and total ICP for 7.5 V (P < .001 vs controls). Combined treatment returned reduced ICP/MAP and total ICP responses, while partial amelioration was observed after Levothyroxine and Sustanon treatment alone. In vitro endothelium-dependent acetylcholine-caused relaxation at 1 nM (P < .001 vs controls), electrical field stimulation-induced relaxant response at 20 Hz (P < .001 vs controls) and sildenafil-induced relaxation at 10 µM (P < .05 vs controls) were decreased in the CC strips from hypothyroid rats. The treatment with Levothyroxine and Sustanon increased the reduction in relaxation responses. Decreased in the expressions of eNOS and nNOS and increased in the expressions of PDE5 in the hypothyroid group were restored by Levothyroxine and Sustanon. Mono-treatment partially enhanced reduced smooth muscle mass while combined therapy completely recovered. Conclusions These results show that hypothyroidism adversely affects T levels and erectile responses via decreasing neurogenic and endothelium-dependent relaxation responses which were reversed by the combination therapy with Levothyroxine and Sustanon. Even though the associated alterations in T levels, ED and its reversibility indicate an exact role of thyroid hormones in the erection physiology. Disclosure No

Adipose-Derived Stem Cells from Type 2 Diabetic Rats Retain Positive Effects in a Rat Model of Erectile Dysfunction.

Erectile dysfunction is a common complication associated with type 2 diabetes mellitus (T2DM) and after prostatectomy in relation to cancer. The regenerative effect of cultured adipose-derived stem cells (ASCs) for ED therapy has been documented in multiple preclinical trials as well as in recent Pase 1 trials in humans. However, some studies indicate that diabetes negatively affects the mesenchymal stem cell pool, implying that ASCs from T2DM patients could have impaired regenerative capacity. Here, we directly compared ASCs from age-matched diabetic Goto–Kakizaki (ASCGK) and non-diabetic wild type rats (ASCWT) with regard to their phenotypes, proteomes and ability to rescue ED in normal rats. Despite ASCGK exhibiting a slightly lower proliferation rate, ASCGK and ASCWT proteomes were more or less identical, and after injections to corpus cavernosum they were equally efficient in restoring erectile function in a rat ED model entailing bilateral nerve crush injury. Moreover, molecular analysis of the corpus cavernosum tissue revealed that both ASCGK and ASCWT treated rats had increased induction of genes involved in recovering endothelial function. Thus, our finding argues that T2DM does not appear to be a limiting factor for autologous adipose stem cell therapy when correcting for ED.

ITRAQ-based identification of differentially expressed proteins in ED rat model induced by cold stress combined with environmental oestrogen.

We aim to identify serum biomarkers and key proteins of erectile dysfunction (ED) induced by cold stress combined with environmental oestrogen through iTRAQ combined with LC-MS/MS. ED rat model was established by using oestrogen-like feed and cold stimulation. Differentially expressed proteins in serum were screened and analysed with iTRAQ combined with LC-MS/MS and IPA bioanalysis platform. We found that 35 proteins were differentially expressed in the ED group, including 31 upregulated and four downregulated proteins. The molecular functions of differential proteins mainly involved the activation of endopeptidase inhibitor and regulator. The main pathways involved were the FXR/RXR activation signalling pathway and the acute phase response signalling pathway. Furthermore, three protein markers related to ED were obtained, namely C-reactive protein (CRP), T-kininogen 1 and apolipoprotein Protein H (APOH). ELISA results showed that compared with the control group, proteins of HPX, TTR, APOH, RBP4, CRP and ORM1 were significantly upregulated, but ANGPT1 protein was significantly downregulated in the serum of the ED group (p<0.05). Conclusively, the candidate serum markers for ED induced by cold stress combined with environmental hormones were obtained. Our results indicate that inflammation and vascular endothelial function changes may play a key role in the occurrence and development of ED.

Establishing a Thermal Imaging Technology (IRT) Based System for Evaluating Rat Erectile Function.

IntroductionMeasurement of intra-cavernous pressure (ICP) is an internationally recognized method to evaluate erectile function of animals, however, this process is invasive, destructive, and cannot be repeated, leading to a daunting challenge for monitoring the changes in erectile function throughout the whole treatment duration.AimTo verify whether infrared ray thermography technology based system could be a good substitution of ICP for evaluating rat erectile function.MethodsA novel thermal image-based method, infrared ray thermography technology (IRT) was employed to monitor erectile function in erectile dysfunction (ED) rats. To detect the sensitivity and specificity of this new technology, 4 ED rat models (Diabetic, nerve-injury, vascular-injury and aged ED models) were established and subjected to both ICP and IRT test.OutcomesStatistical comparisons were done to test the effectiveness of this new way for detecting and dynamically monitoring erectile function.ResultsBased on the data curves obtained from ICP and IRT, the IRT showed a similar trend (including peak value, climbing speed) as that of ICP. IRT is considered as a precise way to monitor the real-time changes of erectile function in all ED rat models. The AUC of peak temperature detected by IRT in DMED, aged ED, vascular-injury ED, the nerve-injury ED and total ED rat models were 0.9811,0.9836,0.9893,0.9989 and 0.9882, respectively. Meanwhile, the AUC of temperature climbing rate were 0.6486,0.8357,0.9184,0.8675and 0.8168.Also,it is a non-invasive process of dynamically monitoring erectile function of a same rat at different time points (before and after drug intervention). The data showed that the real-time recovery by tadalafil was obtained by IRT methods even after treatment for only 2 weeks in the diabetic ED (DMED) rat model.ConclusionA novel noninvasive method for monitoring erectile function in rat ED models was established, and can replace or supplement ICP test.Liu S, Zhao Z, Wang Z et al. Establishing a Thermal Imaging Technology (IRT) Based System for Evaluating Rat Erectile Function. Sex Med 2022;10:100475.

In vitro and in vivo antioxidant potentials of the methanolic crude extract from Inula glomerata Oliv. &amp; Hiern (Asteraceae) and Salacia kraussii (Harv.) Harv (Celastraceae)

Reactive oxygen species are implicated in multiple pathological conditions including erectile dysfunction. This study evaluated the in vitro and in vivo antioxidant potential of the methanolic extracts of Inula glomerata and Salacia kraussii. The plant materials were pulverized and extracted with methanol. The phytochemical analysis, ability of the crude extracts to scavenge free radicals (ABTS, DPPH, NO.) in vitro as well as the total phenolic and flavonoid contents was investigated. In vivo, antioxidant potentials of the crude extracts (50/250 mg/kg body weight) were determined in an erectile dysfunction rat model. The phytochemical analysis revealed that both plants contain flavonoids, tannins, terpenoids, and alkaloids. The crude extracts at varying degree of efficiency, scavenged ABTS and DPPH radicals. The crude extracts at low concentrations (50 mg/kg b.w) significantly (p&lt;0.05) diminished the level of malondialdehyde, augmented catalase activities and elevated glutathione levels. However, SOD activities were significantly boosted in a dose-dependent manner by the crude extracts. Therefore, I. glomerata and S. kraussii possess antioxidant properties, hence, can serve as a therapeutic modality in the treatment of oxidative stress-induced erectile dysfunction.

Exosomes derived from miR-301a-3p-overexpressing adipose-derived mesenchymal stem cells reverse hypoxia-induced erectile dysfunction in rat models

BackgroundErectile dysfunction (ED) has often been observed in patients with obstructive sleep apnea (OSA). Research on adipose-derived mesenchymal stem cell (ADSC)-derived exosomes has shown that they have significant therapeutic effects in many diseases including ED.MethodsIn this study, ED was induced in Sprague Dawley (SD) rats using chronic intermittent hypoxia (CIH) exposure. CIH-mediated influences were then measured in the corpus cavernous smooth muscle cells (CCSMCs).ResultsOur data showed that miR-301a-3p-enriched exosome treatment significantly recovered erectile function in rats and CCSMCs by promoting autophagy and inhibiting apoptosis. The treatment also significantly recovered the level of alpha smooth muscle actin (α-SMA) in rats and CCSMCs. Bioinformatics predicted that phosphatase and tensin homolog (PTEN) and Toll-like receptor 4 (TLR4) might be targets of miR-301a-3p.ConclusionsOur results indicate that PTEN-overexpression vectors or TLR4-overexpression vectors reverse the therapeutic effects achieved by miR-301a-3p in CCSMCs indicating that PTEN/hypoxia-inducible factor-1 alpha (HIF-1α) and TLR4 signaling pathways play key roles in the progression of ED. The findings in this study suggest that miR-301a-3p should be considered a new therapeutic target for treating ED associated with OSA.

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.

Current Status and Prospects in the Treatment of Erectile Dysfunction by Adipose-Derived Stem Cells in the Diabetic Animal Model.

Erectile dysfunction (ED) is defined as the persistent inability to achieve and maintain an erection status sufficient to permit satisfactory sexual performance. Current evidence suggests that diabetes mellitus erectile dysfunction (DMED) is one of the leading causes of ED which remains a difficult condition to manage because of its complicated pathophysiological mechanisms. Recently, stem cell therapies have been added to the therapeutic treatment options for ED. Stem cells derived from adipose tissue are now considered an alternative approach to DMED with preliminary studies demonstrating their capability of promoting endothelial cell, smooth muscle cell, and cavernous nerve regeneration. We will review the epidemiology and pathophysiology of ED, rat models, and adipose-derived stem cell (ASC) therapy and its effects. The relevant literature and contemporary data, using keywords "adipose-derived stem cells and diabetes erectile dysfunction," were reviewed. The main outcome measure was the evidence supporting the association between ASCs, diabetes ED, and rat model. ASCs can restore erectile function of DMED rats by promoting vascularization and neuralization of corpus cavernosum. They can also inhibit fibrosis and inflammation and protect smooth muscle cells. ASCs have achieved satisfactory therapeutic effects in rat models of ED, but effectiveness and safety of their application in clinical research remain to be determined. Yan H, Ding Y, Lu M. Current Status and Prospects in the Treatment of Erectile Dysfunction by Adipose-Derived Stem Cells in the Diabetic Animal Model. Sex Med Rev 2020;8:486-491.

PRESERVATION OF ERECTILE FUNCTION BY STATINS IN A RAT MODEL OF ERECTILE DYSFUNCTION INDUCED BY HYPERCHOLESTEROLEMIA

Background and objectiveTo assess the effects of hypercholesterolemia (HC) on the quality of erections and to evaluate the effects of stain therapy in a rat model of erectile dysfunction (ED) induced by HC.Material and methodsSprague&ndash;Dawley rats were randomly divided into three groups (n=12 in each): control, HC, and HC with simvastatin treatment (HC+SS). The control was fed a normal chow diet, and the HC and HC+SS were fed a high-fat and high-cholesterol diet for 12 weeks. The HC+SS received simvastatin once daily via oral gavage for 12 weeks. Subsequently, the intraperitoneal glucose tolerance test (IPGTT), intracavernous pressure and mean arterial pressure, lipid profiles, expression of endothelial nitric oxide synthase (eNOS) and neuronal nitric oxide synthase (nNOS), oxidative stress (8-hydroxy-2-deoxyguanosine, 8-OHdG level), serum testosterone levels, and the ratio of collagen fibers (CF) and smooth muscle (SM) were evaluated in the serum and corpora tissue.ResultsIPGTT was not different among all groups. The HC showed markedly lower erectile parameters than the control. In contrast, the HC+SS showed preserved erectile function, improved lipid profiles, increased eNOS and nNOS, decreased oxidative stress, and minimized change in SM/CF ratio. ConclusionsOur results suggest that oxidative stress damage by HC may cause ED and that statin therapy may have beneficial effects on preserving erectile function by improving lipid profiles and minimizing damage caused by oxidative stress.

102 Adipose-derived Stem Cells (ASCs) Combined with Control-released DF-PEG/GCS Hydrogel Scaffold Restore the Erectile Function in a Diabetes Rat Model

To investigate the feasibility of seeding adipose-derived stem cells (ASCs) onto a control-released hydrogel scaffold, Telechelic difunctional poly/ Glycol chitosan (DF-PEG/GCS), and then inject the scaffold into corpus cavernosum for erectile function restoration in a diabetes mellitus erectile dysfunction rat model and investigate the underlying mechanism. Fourty-eight 8-week-old male Sprague-Dawley rats were randomly divided into 6 groups, including the Normal (n=8) group, PBS (n=8) group, ASCs (n=8) group, G+ASCs group (n=8), D+ASCs group (n=8) and H+ASCs group (n=8). For in vivo studies, rats with streptozocin-induced DM were intravenously injected with PBS, ASCs or ASCs seeded on GCS, DF-PEG or DF-PEG/GCS Hydrogel and rats in Normal group were injected with an equal volume of vehicle solution. Twelve weeks later, the intracavernosal pressure (ICP) and mean arterial pressure (MAP) were measured to assess erectile function, and penile tissues were harvested for further evaluation. A significant fall in the ICP was obvious in diabetic rats compared with normal rats. After rats with diabetes were treated with ASCs seeded DF-PEG/GCS Hydrogel, the ICP/MAP ratio significantly increased compared with that in the PBS group. Partial restoration of the ICP was observed in ASCs group. The CD31, a-SMA and NEUN expression levels were considerably decreased in the corpus cavernosum tissue of the PBS group compared with the Normal group (P < 0.05;). Twelve weeks after treatment with ASCs, expression levels of CD31, NEUN and a-SMA were significantly increased compared with those treated with PBS (P < 0.05). There was no significant difference between the G+ASCs, D+ASCs and ASCs groups. Western blot analysis was performed to determine the expression levels of bioactive factors NGF, BDNF and VEGF. Densitometric analysis of the band intensity revealed that in all cases, the decrease in expression of cell growth factors (NGF, BDNF and VEGF) as a result of diabetes in the rats was partially recovered by ASCs, and further restored by ASCs seeded DF-PEG/GCS hydrogel scaffold. Changes in expression of the inflammatory cytokines TNF-α, IL-6 and IL-1β in the penile tissue were investigated by real-time PCR. In all cases, the sharp increase in the expression of inflammatory factors in the diabetic rats compared with the normal rats, was attenuated by ASCs and further decreased by ASCs seeded hydrogel scaffold (P < 0.05 for TNF-α, IL-6 and IL-1β).

Non-Invasive Cell Tracking with Brighter and Red-Transferred Luciferase for Potential Application in Stem Cell Therapy

This study investigated the safety of a novel cell-labeling technology with mKATE and Renilla reniformis luciferase (mKATE-renLUC) and assessed the efficacy on tracking implanted human placental stromal cells (PSC) in an erectile dysfunction (ED) animal model. Human PSC were labeled with mKATE-renLUC using a lentivirus. Cell viability, apoptosis, proliferation, migration, surface marker expression and differentiation potential of the labeled PSC were evaluated and compared with non-labeled PSC. The paracrine profile of labeled cells was examined using an angiogenesis protein array. The brightness and duration of labeled cells with different densities were evaluated. An ED rat model was established and labeled PSC were injected into cavernosal tissue of the penis. The migration and distribution of transplanted PSC were monitored using an IVIS imaging system in real time. Implanted PSC were identified in isolated tissues via detection of mKATE fluorescence. The cell viability, morphology, proliferation, migration, surface marker expression and differentiation potential of mKATE-renLUC-labeled PSC were similar to those of non-labeled cells in vitro (no statistical difference p>0.05). Similar expressions of trophic factors were found between labeled and non-labeled PSC. The migration and distribution of PSC expressing renLUC were tracked in vivo using IVIS imaging system. mKATE-positive PSC were detected in penile, kidney, prostate and hepatic tissues using histological methods. This labeling technology provides a safe and effective cell-tracking approach with a brighter fluorophore and codon-optimized luciferase.