Electrical Stimulation Of Cavernous Nerve Research Articles

(274) BMP2 Restores Erectile Dysfunction Through Neurovascular Regeneration and Fibrosis Reduction in Diabetic Mice

Abstract Introduction The odds of erectile dysfunction (ED) are thrice more prevalent in diabetes. Severe peripheral vascular and neural damage in diabetic patients responds poorly to PDE5 inhibitors. However, bone morphogenetic protein 2 (BMP2) is known to be involved in angiogenesis. Objective To assess the efficacy of BMP2 stimulating angiogenesis and augmenting nerve regeneration in a mouse model of diabetic-induced erectile dysfunction. Methods The induction of diabetes mellitus was done by streptozotocin (STZ, 50 mg/kg daily) administered intraperitoneally for five successive days to male C57BL/6 mice that were 8 weeks old. Eight weeks post inductions, animals were allocated to one of five groups: a control group, an STZ-induced diabetic mouse group receiving two intra-cavernous 20 μL PBS injections, or one of three BMP2 groups administered two injections of BMP2 protein (1μg, 5 μg, or 10 μg) diluted in 20 μL of PBS with three days interval between the first and second injection. The erectile functions were assessed two weeks after PBS or BMP2 protein injections by recording the ICP (intracavernous pressure) through cavernous nerve electrical stimulation. Angiogenic activities also nerve regenerating effects of BMP2 were determined in penile tissues, aorta, vena cava, the main pelvic ganglions, the dorsal roots, and from the primary cultured MCECs (mouse cavernous endothelial cells). Moreover, fibrosis-related factors protein expressions were evaluated by western blot. Results Erectile function recovery to 81% of the control value in diabetic mice was found with intra-cavernous BMP2 injection (5 μg/20 μL). Pericytes and also endothelial cells were extensively restored. It was confirmed that angiogenesis was promoted in the corpus cavernosum of diabetic mice treated with BMP2 through increased ex vivo sprouting of aortic rings, vena cava and penile tissues, and migration and tube formation of MCECs. BMP2 protein enhanced cell proliferation and reduced apoptosis in MCECs and penile tissues, and promoted neurite outgrowth in major pelvic ganglia and dorsal root ganglia under high-glucose conditions. Furthermore, BMP2 suppressed fibrosis by reducing MCEC fibronectin, collagen 1, and collagen 4 levels under high-glucose conditions. Conclusions BMP2 modulates neurovascular regeneration and inhibits fibrosis to revive the mice's erection function in diabetic conditions. Our findings propose that the BMP2 protein represents a novel and promising approach to treating diabetes-related erectile dysfunction (ED). Disclosure No.

Dimethyl fumarate ameliorates erectile dysfunction in bilateral cavernous nerve injury rats by inhibiting oxidative stress and NLRP3 inflammasome-mediated pyroptosis of nerve via activation of Nrf2/HO-1 signaling pathway

ObjectiveTo investigate the therapeutic potential of dimethyl fumarate (DMF) in improving erectile function of bilateral cavernous nerve injury (BCNI) rats, along with elucidating its underlying mechanisms. MethodsA BCNI rat model was established by clamping bilateral cavernous nerve (CN). DMF was given by gavage at low (20 mg/kg/day) and high (40 mg/kg/day) dosages for a duration of 4 weeks. Erectile function was assessed by electrical stimulation of CN. Penis and CN tissues were collected for subsequent analysis. Additionally, PC-12 cell line was used to verify the mechanism of DMF in vitro. Nfe2l2 or Ho-1 gene knockdown PC-12 cell lines were constructed by lentiviral transfection, respectively. A damaged cell model was induced using H2O2. And then molecular biological methods were employed to analyze cellular molecules and proteins. ResultsDMF administration for 4 weeks led to improvements in erectile function, reduced fibrosis of penis corpus cavernosum in BCNI rats. The morphology of CN was improved and the number of nerve fibers increased. Furthermore, the levels of nNOS, NO, and cGMP were increased, while Ca2+ was decreased in penis corpus cavernosum. Notably, the levels of ROS, 3-NT and NLRP3 inflammasomes production were reduced, alongside increased expression of Nrf2 and HO-1 proteins in the dorsal penile nerve (DPN) and CN. In vitro, DMF increased cell viability, reduced ROS level, promoted SOD, diminished 3-NT, MDA and DNA damage markers, and inhibited the activation of NLRP3 inflammasomes in H2O2 induced PC-12 cells. Nfe2l2 knockdown and Ho-1 knockdown significantly attenuated the protective effect of DMF, respectively. Furthermore, inhibition of ROS production by N-acetylcysteine led to a reduction in NLRP3 inflammasome activation in H2O2 induced PC-12 cells. ConclusionsDMF improved erectile function of BCNI rats by protecting nerves through inhibiting oxidative stress and the activation of NLRP3 inflammasome-mediated pyroptosis via activation of Nrf2/HO-1 pathway.

BMP2 restores erectile dysfunction through neurovascular regeneration and fibrosis reduction in diabetic mice.

The odds of erectile dysfunction are three times more prevalent in diabetes. Severe peripheral vascular and neural damage in diabetic patients responds poorly to phosphodiesterase-5 (PDE5) inhibitors. However, bone morphogenetic protein 2 is known to be involved in angiogenesis. To assess the efficacy of bone morphogenetic protein 2 in stimulating angiogenesis and augmenting nerve regeneration in a mouse model of diabetic-induced erectile dysfunction. The induction of diabetes mellitus was performed by streptozotocin (50mg/kg daily) administered intraperitoneally for 5 successive days to male C57BL/6 mice that were 8weeks old. Eight weeks post-inductions, animals were allocated to one of five groups: a control group, a streptozotocin-induced diabetic mouse group receiving two intracavernous 20μL phosphate-buffered saline injections, or one of three bone morphogenetic protein 2 groups administered two injections of bone morphogenetic protein 2 protein (1, 5, or 10μg) diluted in 20μL of phosphate-buffered saline within a 3-day interval between the first and second injections. The erectile functions were assessed 2 weeks after phosphate-buffered saline or bone morphogenetic protein 2 protein injections by recording the intracavernous pressure through cavernous nerve electrical stimulation. Angiogenic activities and nerve regenerating effects of bone morphogenetic protein 2 were determined in penile tissues, aorta, vena cava, the main pelvic ganglions, the dorsal roots, and from the primary cultured mouse cavernous endothelial cells. Moreover, fibrosis-related factor protein expressions were evaluated by western blotting. Erectile function recovery to 81% of the control value in diabetic mice was found with intracavernous bone morphogenetic protein 2 injection (5μg/20μL). Pericytes and endothelial cells were extensively restored. It was confirmed that angiogenesis was promoted in the corpus cavernosum of diabetic mice treated with bone morphogenetic protein 2 through increased ex vivo sprouting of aortic rings, vena cava and penile tissues, and migration and tube formation of mouse cavernous endothelial cells. Bone morphogenetic protein 2 protein enhanced cell proliferation and reduced apoptosis in mouse cavernous endothelial cells and penile tissues, and promoted neurite outgrowth in major pelvic ganglia and dorsal root ganglia under high-glucose conditions. Furthermore, bone morphogenetic protein 2 suppressed fibrosis by reducing mouse cavernous endothelial cell fibronectin, collagen 1, and collagen 4 levels under high-glucose conditions. Bone morphogenetic protein 2 modulates neurovascular regeneration and inhibits fibrosis to revive the mouse erection function in diabetic conditions. Our findings propose that the bone morphogenetic protein 2 protein represents a novel and promising approach to treating diabetes-related erectile dysfunction.

MP27-10 CRISPRA-ENGINEERED ADIPOSE-DERIVED STEM CELLS IMPROVE ERECTILE DYSFUNCTION IN DIABETIC RATS

You have accessJournal of UrologyCME1 Apr 2023MP27-10 CRISPRA-ENGINEERED ADIPOSE-DERIVED STEM CELLS IMPROVE ERECTILE DYSFUNCTION IN DIABETIC RATS Wenchao Xu, Hao Li, Taotao Sun, Jiaxin Wang, Tao Wang, and Jihong Liu Wenchao XuWenchao Xu More articles by this author , Hao LiHao Li More articles by this author , Taotao SunTaotao Sun More articles by this author , Jiaxin WangJiaxin Wang More articles by this author , Tao WangTao Wang More articles by this author , and Jihong LiuJihong Liu More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000003255.10AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Due to the high incidence of diabetes mellitus (DM) and poor response to phosphodiesterase type 5 inhibitor in DM-induced erectile dysfunction (DMED) patients, new therapeutic strategies for DMED are needed. Adipose-derived stem cells (ADSCs) transplantation is considered a promising treatment for DMED but is limited by poor survival and efficacy after transplantation. This study explored the therapeutic effect of CRISPRa-engineered ADSCs overexpressing GPX4 (GPX4-ADSCs) on erectile function in diabetic rats. METHODS: Thirty-two SD rats were randomly divided into four groups: the control, DMED, ADSCs, GPX4-ADSCs groups. Among them, the control group rats were non-diabetic rats, and the latter four groups were all diabetic ED rats of the same age. CRISPRa system was used to overexpress GPX4 in ADSCs. The GPX4-ADSCs group was treated with intracavernous injections of GPX4-ADSCs, while the ADSCs group was treated with ADSCs. Four weeks after transplantation, intracavernous pressure (ICP) and mean arterial pressure (MAP) were measured by electrical stimulation of cavernous nerve, and penile tissue was collected for subsequent testing. RESULTS: ADSCs and GPX4-ADSCs transplantation did not affect the weight or the glucose levels of diabetic rats. Although both the ADSCs group and the GPX4-ADSCs group improved erectile function in diabetic rats, the GPX4-ADSCs group had better improvement than the ADSCs group. In the DMED group, oxidative levels in the corpus cavernosum increased, PI3K/AKT/eNOS signal expression was down-regulated, Caspase3 activity was enhanced, and TGFβ1-Smad2/3-Collagen IV pathway was upregulated, showing obvious oxidative stress, apoptosis and fibrosis. ADSCs and GPX4-ADSCs can reverse these changes caused by diabetes to some extent, and GPX4-ADSCs are more effective than ADSCs. CONCLUSIONS: The CRISPRa-engineered ADSCs overexpressing GPX4 had stronger efficacy in improving erectile function than ADSCs, and the mechanism may involve reducing oxidative stress, apoptosis, and fibrosis, which provides a new strategy for the treatment of DMED. Source of Funding: This research was funded by grants from the National Natural Science Foundation of China (No. 82001536 and No.81873831) and the Fundamental Research Funds for the Central Universities (HUST: YCJJ202201021) © 2023 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 209Issue Supplement 4April 2023Page: e366 Advertisement Copyright & Permissions© 2023 by American Urological Association Education and Research, Inc.MetricsAuthor Information Wenchao Xu More articles by this author Hao Li More articles by this author Taotao Sun More articles by this author Jiaxin Wang More articles by this author Tao Wang More articles by this author Jihong Liu More articles by this author Expand All Advertisement PDF downloadLoading ...

Nitro-oleic acid (NO2-OA) ameliorates erectile dysfunction in a rat model of diabetes mellitus via modulation of fibrosis, inflammation and autophagy.

Inflammation, fibrosis and autophagy represent closely related factors associated with the pathogenesis of diabetes mellitus erectile dysfunction (DMED). In this study, the therapeutic effect of nitro-oleic acid (NO2-OA) in a streptozotocin-induced rat model of DMED was evaluated. Sixty rats were randomly divided into four groups: control, DMED, DMED + Vehicle and DMED + NO2-OA. DMED was induced by intraperitoneal injection of streptozotocin in male rats. Blood glucose and body weight were measured every 2 weeks. After 4 weeks of NO2-OA treatment, erectile function was measured by electrical stimulation of cavernous nerve (CN). Western blotting, quantitative real-time polymerase chain reaction (qRT-PCR), enzyme-linked immunosorbent assay (ELISA), immunofluorescence and Masson's trichrome staining were used to verify the related factors and protein expression levels. We found that NO2-OA could significantly increase erectile pressure in the corpus cavernosum of DMED rats. Results of western blot, confocal immunofluorescence and qRT-PCR assays revealed that NO2-OA significantly reduced inflammatory factors and the expression of nuclear factor kappa B (NF-κB). In addition, Masson staining results indicated that NO2-OA significantly reduced the display of fibrotic tissue in the corpus cavernosum. These beneficial effects may be related to reductions in the expression of transforming growth factor-β1 (TGF-β1) and connective tissue growth factor (CTGF) and the increase in the expression of α-smooth muscle actin (α-SMA). Finally, NO2-OA treatment increased the expression of the autophagy marker, LC3, while P62 was decreased, effects suggesting that one of the underlying mechanisms of NO2-OA may involve an activation of the PI3K/AKT/mTOR pathway to enhance the capacity for autophagy within this tissue. NO2-OA enhances erectile function within a rat model of DMED by inhibiting inflammation and fibrosis along with activating autophagy.

Intracavernosal onabotulinumtoxin a exerts a synergistic pro-erectile effect when combined with sildenafil in spontaneously hypertensive rats

<h3>Objectives</h3> Our overall aim was to provide experimental pharmacological evidence for the use of intracavernosal (ic) onabotulinumtoxinA alone or in combination with phosphodiesterase type 5 inhibitors for difficult-to-treat erectile dysfunction. We thus compared the effects of botulinum toxin A alone and botulinum toxin A combined with phosphodiesterase type 5 inhibitor, and a placebo treatment. <h3>Methods</h3> Erectile function was evaluated following cavernous nerve electrical stimulation at different frequencies in 4 groups (n = 8 / group) of anesthetized, spontaneously hypertensive rats. Rats were treated by onabotulinumtoxinA 10U or saline ic 1 week prior to erectile function testing and sildenafil (0.3 mg/kg) or saline iv 4 minutes prior to testing. Frequency-response curves were compared with a two-way ANOVA. <h3>Results</h3> Intracavernosal pressure/mean arterial pressure ratios were significantly increased by sildenafil and onabotulinumtoxinA ic versus the control condition. OnabotulinumtoxinA 10U ic combined with sildenafil significantly potentiated erectile responses. Area under the curve/mean arterial pressure ratio increased by 19% with sildenafil, by 15% with onabotulinumtoxinA ic and by 58% with the combined treatment. <h3>Conclusions</h3> Both onabotulinumtoxinA ic, and sildenafil, significantly improved erectile responses in spontaneously hypertensive rats, however the effect was greatly amplified when the treatments were combined. These results support further studies into the mechanisms behind the pro-erectile effect of BTX-A ic, as well as multicenter randomized control trials to evaluate the safety and efficacy of BTX-A combined with sildenafil for difficult-to-treat ED. <h3>Conflicts of Interest</h3> No conflict of interest

STIM/Orai Inhibition as a Strategy for Alleviating Diabetic Erectile Dysfunction Through Modulation of Rat and Human Penile Tissue Contractility and in vivo Potentiation of Erectile Responses

Stromal interaction molecule (STIM)/Orai calcium entry system appears to have a role in erectile dysfunction (ED) pathophysiology but its specific contribution to diabetic ED was not elucidated. To evaluate STIM/Orai inhibition on functional alterations associated with diabetic ED in rat and human penile tissues and on in vivo erectile responses in diabetic rats. Rat corpus cavernosum (RCC) strips from nondiabetic (No DM) and streptozotocin-induced diabetic (DM) rats and human penile resistance arteries (HPRA) and corpus cavernosum (HCC) from ED patients undergoing penile prosthesis insertion were functionally evaluated in organ chambers and wire myographs. Erectile function in vivo in rats was assessed by intracavernosal pressure (ICP) responses to cavernous nerve electrical stimulation (CNES). Expression of STIM/Orai elements in HCC was determined by immunofluorescence and immunoblot. Functional responses in RCC, HCC and HPRA and STIM/Orai protein expression in HCC. In vivo erectile responses to CNES. Inhibition of Orai channels with YM-58483 (20 µM) significantly reduced adrenergic contractions in RCC but more effectively in DM. Thromboxane-induced and neurogenic contractions were reduced by STIM/Orai inhibition while defective endothelial, neurogenic and PDE5 inhibitor-induced relaxations were enhanced by YM-58483 (10 µM) in RCC from DM rats. In vivo, YM-58483 caused erections and attenuated diabetes-related impairment of erectile responses. YM-58483 potentiated the effects of PDE5 inhibition. In human tissues, STIM/Orai inhibition depressed adrenergic and thromboxane-induced contractions in ED patients more effectively in those with type 2 diabetes. Diabetes was associated with increased expression of Orai1 and Orai3 in ED patients. Targeting STIM/Orai to alleviate diabetes-related functional alterations of penile vascular tissue could improve erectile function and potentiate therapeutic effects of PDE5 inhibitors in diabetic ED. Improving effects of STIM/Orai inhibition on diabetes-related functional impairment was evidenced in vitro and in vivo in an animal model and validated in human tissues from ED patients. Functional findings were complemented with expression results. Main limitation was low numbers of human experiments due to limited human tissue availability. STIM/Orai inhibition alleviated alterations of functional responses in vitro and improved erectile responses in vivo in diabetic rats, potentiating the effects of PDE5 inhibition. STIM/Orai inhibition was validated as a target to modulate functional alterations of human penile vascular tissue in diabetic ED where Orai1 and Orai3 channels were upregulated. STIM/Orai inhibition could be a potential therapeutic strategy to overcome poor response to conventional ED therapy in diabetic patients. Sevilleja-Ortiz A, El Assar M, García-Gómez B, et al. STIM/Orai Inhibition as a Strategy for Alleviating Diabetic Erectile Dysfunction Through Modulation of Rat and Human Penile Tissue Contractility and in vivo Potentiation of Erectile Responses. J Sex Med 2022;19:1733-1749.

Intracavernosal OnabotulinumtoxinA Exerts a Synergistic Pro-Erectile Effect When Combined With Sildenafil in Spontaneously Hypertensive Rats

Botulinum toxin A (BTX-A) has a variety of uses in medicine. Some evidence suggests that intracavernosal (ic) BTX-A injection administered in addition to phosphodiesterase type 5 inhibitors (PDE5-Is) could effectively treat erectile dysfunction (ED) in insufficient responders to PDE5-Is. To provide experimental pharmacological evidence for the use of onabotulinumtoxinA ic alone or in combination with PDE5-Is for difficult-to-treat ED. We thus compared the effects of BTX-A ic alone and BTX-A ic combined with PDE5-I iv, and a placebo treatment ic or iv. Erectile function was evaluated following cavernous nerve electrical stimulation (6 V, 1-millisecond pulse, 45-second duration) at different frequencies (0, 2, 3, 4, 5, 7.5, and 10 Hz) in 4 groups (n = 8 / group) of anesthetized, spontaneously hypertensive rats, a robust animal model of ED of vascular origin. Rats were treated by onabotulinumtoxinA 10U or saline ic 1 week prior to erectile function testing and sildenafil (0.3 mg/kg) or saline iv 4 minutes prior to testing. Frequency-response curves were compared with a 2 way ANOVA. Both onabotulinumtoxinA ic, and sildenafil iv significantly improved erectile responses in spontaneously hypertensive rats, however the effect was greatly amplified when the treatments were combined. Intracavernosal pressure and/or mean arterial pressure ratios were significantly increased by sildenafil and onabotulinumtoxinA ic versus the control condition. OnabotulinumtoxinA 10U ic combined with sildenafil iv significantly potentiated erectile responses. Area under the curve and/or mean arterial pressure ratio increased by 19% with sildenafil iv, by 15% with onabotulinumtoxinA ic and by 58% with the combined treatment following cavernous nerve electrical stimulation at 6V, 1 ms, 10 Hz: these stimulation parameters elicited the maximal erectile response. These data provide a pharmacological rationale for the combined administration of onabotulinumtoxinA ic and sildenafil iv since the effects of both treatments were potentiated when their administration was combined. First evidence of a synergistic pro-erectile effect of BTX-A combined with PDE5-I, however the mechanism behind the pro-erectile effect of BTX-A ic remains hypothetical. These results support further studies into the mechanisms behind the pro-erectile effect of BTX-A ic, as well as multicenter randomized control trials to evaluate the safety and efficacy of BTX-A ic combined with sildenafil for difficult-to-treat ED.

Diabetes associated with hypertension exacerbated oxidative stress-mediated inflammation, apoptosis and autophagy leading to erectile dysfunction in rats.

Diabetes or hypertension contributes to erectile dysfunction (ED). We hypothesized that excess reactive oxygen species (ROS) production evoked by diabetes combined with hypertension may further suppress endothelial nitric oxide (NO) expression/activity and promote oxidative stress in the ED penis. Twenty-four adult male Wistar Kyoto rats (WKY) and spontaneously hypertensive rats (SHR) were divided into four groups: normal WKY, diabetic WKY, normal SHR and diabetic SHR. Intraperitoneal streptozotocin (65 mg/kg) was applied to induce type I diabetes. After 4-week diabetes and/or hypertension induction, we determined the intra-cavernous pressure (ICP) using electrical stimulation of cavernous nerves, intra-cavernosum NO amount using an electrochemical NO probe, and blood ROS using an ultrasensitive chemiluminescence-amplified analyzer. Western blot analysis and immunohistochemistry were used to explore the pathophysiologic mechanisms of inflammation, apoptosis and autophagy in the penis. A novel NO donor, CysaCysd Lu-5 (CCL5, (RCH2CH2S)(R'R"CHCH2S)Fe(NO)2, 1-4 µg), was intravenously administered to these ED rats for evaluating their ICP responses. In the baseline status, the lucigenin- and luminol-amplified blood ROS were significantly enhanced in the diabetic SHR rats vs normal WKY rats. Significantly decreased ICP, eNOS expression and NO amount were found in the normal SHR, diabetic WKY, and diabetic SHR vs normal WKY rats. Intravenous NO donor L-Arginine markedly increased ICP and NO amount, whereas eNOS inhibitor, Nω-Nitro-L-Arginine methyl ester hydrochloride depressed ICP in all four groups. Diabetes and/or hypertension alone increased fibrosis, proinflammatory NF-kB/ICAM-1 expression, mast cell numbers, CD68 expression and infiltration, Caspase 3-mediated apoptosis, Beclin-1/LC3-II-mediated autophagy and mild Nrf-2/HO-1 expression and depressed eNOS expression in the ED penis. The novel NO donor, CCL5, was more efficient than L-arginine to improve diabetes and/or hypertension-induced ED by the significant increase of ICP. Diabetes combined with hypertension synergistically exacerbated ED through enhanced oxidative stress, inflammation, apoptosis and autophagy and depressed eNOS activity and NO production.

Ipidacrine (Axamon), A Reversible Cholinesterase Inhibitor, Improves Erectile Function in Male Rats With Diabetes Mellitus-Induced Erectile Dysfunction.

ABSTRACTSBackgroundManagement of diabetes mellitus-induced erectile dysfunction (DMED) is challenging because of its insufficient responses to phosphodiesterase type 5 inhibitors.AimTo compare the effects of ipidacrine, a reversible cholinesterase inhibitor, and sildenafil on DMED in a rat model of streptozotocin (STZ)-induced diabetes.MethodsErectile dysfunction (ED) caused by STZ-induced diabetes mellitus was modeled in adult male Wistar rats, which were randomized to 4 groups: untreated diabetic rats, sildenafil (5 mg/kg), ipidacrine (3.6 mg/kg) and ipidacrine (6.7 mg/kg). The test drug (ipidacrine), comparator (sildenafil) or control substance (1% starch solution) were administered orally for 5 days or 14 days. Erectile function was assessed by the change in the maximum intracavernous pressure (ICPmax) following cavernous nerve electrical stimulation. The mean arterial pressure (MAP) was recorded, and the ICPmax/MAP ratio was calculated. Sexual behavior, cholinesterase activity and blood testosterone level tests assessed.Main Outcome MeasureThe quantitative value of ICPmax/MAP 14 days after the start of administration of the test drug and the comparison drug.ResultsAnimals with STZ-induced diabetes mellitus showed a significant decrease in ICPmax and ICPmax/MAP ratio compared to the intact control group. When ipidacrine was administered to rats with DMED for 14 days, an increase in these indicators was noted. It was proved that ipidacrine at a dose of 6.7 mg/kg has noninferiority compared to sildenafil on the DMED model. Significant increase in ICPmax compared to STZ-control after electrostimulation of the cavernous nerve was recorded following administration of ipidacrine at a dose of 6.7 mg/kg (P < .05) and sildenafil at a dose 5 mg/kg (P < .05). Neither the test drug, nor the comparator were associated with increase in testosterone levels in blood; as well both drugs did not promote activation of sexual behavior.Clinical ImplicationsIpidacrine may be considered as an effective therapy for DMED but needs to be verified in human investigations.Strengths & LimitationsThe role of ipidacrine, was firstly demonstrated in rats with DMED. However, the results were obtained in animal experiments, and will be further tested in the study of receptor interactions and the determination of cellular targets.ConclusionThis is the first study to show that administration of ipidacrine, the reversible cholinesterase inhibitor, improved erectile function in diabetic rats and these results may be beneficial in further studies using ipidacrine for treatment of DMED, particularly in non-responders to PDE5 inhibitors.Bykov V, Gushchina E, Morozov S, et al. Ipidacrine (Axamon), A Reversible Cholinesterase Inhibitor, Improves Erectile Function in Male Rats With Diabetes Mellitus-Induced Erectile Dysfunction. Sex Med 2022;10:100477.

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.

Exertional heat stroke on fertility, erectile function, and testicular morphology in male rats

The association of exertional heat stroke (EHS) and testicular morphological changes affecting sperm quality, as well as the association of EHS and hypothalamic changes affecting sexual behavior, has yet to be elucidated. This study aimed to elucidate the effects of EHS on fertility, erectile function, and testicular morphology in male rats. Animals were exercised at higher room temperature (36 ℃ relative humidity 50%) to induce EHS, characterized by excessive hyperthermia, neurobehavioral deficits, hypothalamic cell damage, systemic inflammation, coagulopathy, and multiple organ injury. In particular, EHS animals had erectile dysfunction (as determined by measuring the changes of intracavernosal pressure and mean arterial pressure in response to electrical stimulation of cavernous nerves). Rats also displayed testicular temperature disruption, poorly differentiated seminiferous tubules, impaired sperm quality, and atrophy of interstitial Leydig cells, Sertoli cells, and peri-tubular cells in the testicular tissues accompanied by no spermatozoa and broken cells with pyknosis in their seminal vesicle and prostatitis. These EHS effects were still observed after 3 days following EHS onset, at least. Our findings provide a greater understanding of the effect of experimentally induced EHS on masculine sexual behavior, fertility, stress hormones, and morphology of both testis and prostate.

Neutralizing antibody to proNGF rescues erectile function by regulating the expression of neurotrophic and angiogenic factors in a mouse model of cavernous nerve injury.

Radical prostatectomy induces some degree of cavernous nerve injury (CNI) and causes denervation-induced pathologic changes in cavernous vasculature, regardless of the advances in surgical techniques and robotic procedures. The precursor for nerve growth factor (proNGF) is known to be involved in neuronal cell apoptosis and microvascular dysfunction through its receptor p75NTR . To determine the expression of proNGF/p75NTR and the efficacy of proNGF neutralizing antibody (anti-proNGF-Ab) in a mouse model of ED induced by CNI. Age-matched 12-week-old C57BL/6 mice were distributed into three groups: sham group and bilateral CNI group treated with intracavernous injections of PBS (20μL) or of anti-proNGF-Ab (20µg in 20μL of PBS) on days -3 and 0. Two weeks after treatment, erectile function was measured by electrical stimulation of cavernous nerve. Penis tissues from a separate group of animals were harvested for further analysis. We also determined the efficacy of anti-proNGF-Ab on neural preservation in major pelvic ganglion (MPG) ex vivo. We observed increased penile expression of proNGF and p75NTR after CNI. Intracavernous administration of anti-proNGF-Ab increased nNOS and neurofilament expression probably by enhancing the production of neurotrophic factors, such as neurotrophin-3, NGF, and brain-derived neurotrophic factor. Anti-proNGF-Ab preserved the integrity of cavernous sinusoids, such as pericytes, endothelial cells, and endothelial cell-to-cell junctions, possibly by controlling angiogenic factors (angiopoietin-1, angiopoietin-2, and vascular endothelial growth factor) and induced endogenous eNOS phosphorylation in CNI mice. And finally, treatment with anti-proNGF-Ab rescued erectile function in CNI mice. Anti-proNGF-Ab also enhanced neurite sprouting from MPG exposed to lipopolysaccharide. The preservation of damaged cavernous neurovasculature through inhibition of the proNGF/p75NTR pathway may be a novel strategy to treat radical prostatectomy-induced erectile dysfunction.

Extracorporeal Shock Waves Therapy Delivered by Aries Improves Erectile Dysfunction in Spontaneously Hypertensive Rats Through Penile Tissue Remodeling and Neovascularization.

BackgroundLow-intensity extracorporeal shock wave therapy (Li-ESWT) has been reported to improve erectile function in patients with moderate-to-severe erectile dysfunction (ED) or even convert phosphodiesterase type 5 inhibitors nonresponders to responders. ED is highly prevalent in hypertensive patients. The effect of Li-ESWT on an animal model of hypertension-associated ED has not been reported.AimTo investigate the effect of Li-ESWT on hypertension-associated ED and provide plausible mechanisms of action of Li-ESWT on local mechanisms of penile erection.MethodsSpontaneously hypertensive rats (SHRs) in the active group (n = 13) received Li-ESWT at energy flux density 0.06 mJ/mm2 (Aries; Dornier MedTech, Wessling, Germany) twice weekly for 6 weeks. The emitter was set to zero for SHRs in the sham group (n = 12). Erectile function was assessed 4 weeks post-treatment by monitoring intracavernosal pressure (ICP) in response to electrical stimulation of cavernous nerve before and after single dose of 0.3 mg/kg intravenous sildenafil. Cavernosal tissue was then evaluated for collagen/smooth muscle content, neuronal nitric oxide synthase (nNOS), and vascular endothelial factor (CD31) expression.OutcomesErectile function was assessed with ICP, erectile tissue remodeling was studied by smooth muscle/collagen ratio, nNOS and CD31 were semiquantitatively evaluated on cavernosal sections.ResultsThe improvement of ICP parameters was greater in Li-ESWT–treated rats compared with controls with and without sildenafil. Sildenafil led to 20% increase in area under the intracavernosal pressure curve measured during the entire response/mean arterial pressure at 10 Hz in ESWT_SHR + sildenafil compared with ESWT_SHR. The smooth muscle/collagen ratio increased 2.5-fold in Li-ESWT compared with sham. Expression of CD31 tended to be increased whereas nNOS was unchanged.ConclusionsLi-ESWT by Aries may represent an effective noninvasive therapeutic alternative and a relevant add-on therapy to phosphodiesterase type 5 inhibitors for ED in hypertensive patients, and it is suggested that it acts via remodeling of the penile tissue and promoting cavernosal vascularization.Assaly R, Giuliano F, Clement P, et al. Extracorporeal Shock Waves Therapy Delivered by Aries Improves Erectile Dysfunction in Spontaneously Hypertensive Rats Through Penile Tissue Remodeling and Neovascularization. Sex Med 2019;7:441–450

S-nitrosylation of endothelial nitric oxide synthase impacts erectile function

Neuronal and endothelial nitric oxide synthases (nNOS and eNOS respectively) play major roles in generating the nitric oxide bioactivity necessary for erectile function. S-nitrosylation has been shown to regulate NOS activity. The presence of S-nitrosylated NOS in the penis and the impact of NOS S-nitrosylation/denitrosylation on erectile function were examined. S-nitrosylated forms of NOS were identified by biotin-switch assay followed by western blot analysis. Erectile function in S-nitrosoglutathione reductase deficient (GSNO+/-) and null (GSNO-/-) mice were assessed by continuous cavernous nerve electrical stimulation (CCNES). Glutathione ethyl ester (GSHee) was used to manipulate S-nitrosylated NOS levels. Immunohistological and immunofluorescence analyses were used to identify the location of eNOS and GSNO-R in corporal tissue. eNOS and nNOS were S-nitrosylated in unstimulated penises of the mice. CCNES resulted in a time-dependent increase in eNOS S-nitrosylation with peak eNOS S-nitrosylation observed during detumescence. S-nitrosylated nNOS levels were unchanged. Intracorporal injection of GSHee reduced S-nitrosylated eNOS levels, enhancing time to maximum intracorporal pressure (ICP). eNOS and GSNO-R co-localize to the endothelium of the corpus cavernosum in the mouse and the human. ICP measurements obtained during CCNES demonstrate GSNO-R+/- and GSNO-R-/- animals cannot maintain an elevated ICP. Results suggest eNOS S-nitrosylation/denitrosylation is an important mechanism regulating eNOS activity during erectile function. GSNO-R is a key enzyme involved in the eNOS denitrosylation. The increase in eNOS S-nitrosylation (inactivation) observed with tumescence may begin a cycle leading to detumescence. Clinically this may indicate that alterations in the balance of S-nitrosylation/denitrosylation either directly or indirectly contribute to erectile dysfunction.

Treatment with a New Nitric Oxide Donor, a Hybrid Derived from Thalidomide and Hydroxycarbamide 3-(1,3-dioxoisoindolin-2-yl)Benzyl Nitrate, Reverses Priapism in the Sickle Cell Mouse and the Nitric Oxide-Deficient Mouse

▪Introduction: Sickle cell disease (SCD) men display priapism. Previous studies showed that endothelial nitric oxide synthase (eNOS) gene-deficient mice and combined eNOS and neuronal nitric oxide synthase double gene-deficient mice (dNOS-/-) display a priapism phenotype associated with phosphodiesterase-5 (PDE5) downregulation. The Berkeley SCD transgenic mice display features of priapism, and, as in SCD human, this appears to be due to decreased nitric oxide (NO) signaling, which leads to lower PDE5 in the penis. Thus, when an erectile stimulus occurs in vivo, cGMP accumulates into the cavernosal smooth muscle cells, rendering the penile vasculature uncontrollably dilated, penile erection persists (i.e. priapism) as cGMP is not degraded as a consequence of PDE5 downregulation. Moreover, oxidative/nitrosative stress have been associated with priapism in SCD mice. In this study, since decreased NO bioavailability leads to lower PDE5 in penis, we aimed to evaluate the effects of new NO donor, a hybrid derived from thalidomide and hydroxycarbamide, 3-(1,3-dioxoisoindolin-2-yl)benzyl nitrate (4C), on functional and molecular alterations of erectile function in SCD and dNOS-/- mice. We have focused on the dysregulated NO-cGMP-PDE5 pathway and oxidative/nitrosative stress in erectile tissue of SCD and dNOS-/-mice.Methods: Wild type (WT, C57BL/6), Berkeley SCD transgenic and dNOS-/-male mice (3-5 mo old) were treated with compound 4C (100 µmol/kg/day) or its vehicle (20% Cremophor) daily for 3 weeks via intraperitoneal injection. The intracavernous pressure (ICP) was assessed following electrical stimulation of cavernous nerve (CN) in anaesthetized mice. ICP were normalized per mean arterial pressure (MAP). In separate protocols, corpus cavernosum (CC) strips were mounted in isolated organ baths, and the relaxing responses to acetylcholine (ACh; endothelium-dependent response) and sodium nitroprusside (SNP; endothelium-independent response), as well as electrical-field stimulation (EFS; nitrergic relaxation) were obtained in CC strips precontracted with the α1-adrenergic receptor agonist phenylephrine (10 µM).Results: The CN stimulation (4 V) caused increases of ICP/MAP ratio in WT, SCD and dNOS-/- mice. However, half detumescence time (sec) and poststimulated ICP/MAP ratio values after 9 min were (P<0.05) higher in SCD compared to the WT group, indicating that SCD mice display priapism. In dNOS-/- group, maximal ICP/MAP ratio values were higher (P<0.05) compared to WT mice, indicating that dNOS-/- mice also display priapism. Compound 4C treatment reversed the priapism in SCD and dNOS-/- mice. The cumulative addition of ACh (0.001-10 µM) produced concentration-dependent CC relaxations in WT and SCD groups, but the ACh potency (pEC50) value was higher (P<0.05) in CC of SCD compared to WT mice, which was reversed by 4C. Likewise, the nitrergic relaxation induced by EFS was also higher (P<0.05) in SCD mice compared to control mice (2 Hz: 12 ± 3 and 37 ± 9 %, respectively; n=5), which was reversed by 4C treatment. Similarly, SNP (0.01- 10 µM) produced concentration-dependent CC relaxations in WT, SCD and dNOS-/- groups, but, again, the maximal relaxations elicited by this agent were higher (P<0.05) in SCD (93 ± 7%) and dNOS-/- (105 ± 1%) groups compared to WT mice (72 ± 4%), which were fully restored to WT values with compound 4C treatment (n=5-9). PDE5 protein expressions were reduced (P<0.05) by approximately 48% and 35% in penile tissues from SCD and dNOS-/- compared to the WT group, respectively. Compound 4C treatment restored (P<0.05) the protein levels of PDE5 in the penises from SCD and dNOS-/- group. The protein expressions for gp91phox and 3-NT were significantly higher (P<0.05) in erectile tissues from SCD and dNOS-/- compared to the WT group, which were reduced (P<0.05) by 4C treatment (n=5-9). In WT mice, PDE5, gp91phox and 3-NT protein expressions were not affected by 4C, as well as CC relaxations induced by ACh, SNP and EFS.Conclusion: Compound 4C treatment reversed the priapism, as well as enhanced NO-mediated CC relaxations and normalized PDE5 expressions in the penises from SCD and dNOS-/- mice. Moreover, increased gp91phox and 3-nitrotyrosine protein expressions were also decreased by 4C in SCD and dNOS-/-mice. Compound 4C may constitute an additional strategy to prevent priapism in SCD, since SCD-related priapism therapies are few.Financial Support: FAPESP DisclosuresNo relevant conflicts of interest to declare.

Anti-inflammatory and anti-fibrotic effects of annexin1 on erectile function after cavernous nerve injury in rats

The aim of this study was to investigate the effect of the anti-inflammatory and anti-fibrotic actions of ANX1 on erectile function (EF). Forty-eight male Wistar rats were randomly distributed into four equal groups: one group (sham operation-control) and three groups (bilateral cavernous nerve (CN) crush injury). Crush injury groups were treated prior to injury with an intravascular injection of either ANX1 (50 or 100 μg kg-1) or vehicle. EF was assessed by CN electrical stimulation at 2 and 7 days after CN injury with histomorphometric and immunohistochemical analysis. ANX1 demonstrated functional preservation as the increase in intracavernous pressure (ICP). A dose-response relationship regarding the effect on penile tissue was confirmed, and preservation of the penile dorsal nerves and anti-apoptotic effects in the corpus cavernosum (real P-value vs injured control). ANX1 treatment prevented collagen deposition and smooth muscle loss in the penis. ANX1 normalized the expression of vascular endothelial growth factor and decreased tumor necrosis factor-α in the lumen of the blood vessels of the organ. ANX1 proved effective in preserving EF in a rat model of neurogenic erectile dysfunction. ANX1 treatment before CN injury in rats improved erectile recovery, enhanced vascular regeneration and preserved the micro-architecture of the corpus cavernosum. The clinical availability of this compound merits application in penile rehabilitation studies following radical prostatectomy.

Development of Radiation-Induced Erectile Dysfunction Using Conventional Pelvic Radiation Therapy in a Rat Model

Objective: Radiation therapy (RT) is an effective and common treatment for prostate cancer. Although RT-induced erectile dysfunction (ED) is one of the particularly devastating side effects, there is a lack of preclinical studies on RT-induced ED. The aim of this study was to develop a RT-induced ED rat model. Design and Method: 10 adult male Sprague-Dawley (SD) rats were divided into control (n=5) and RT group (n=5). Irradiated rats received external pelvic radiation in a single 24 Gy fraction. After 10 weeks, intracavernous pressure (ICP) and mean arterial pressure (MAP) were measured with cavernous nerve electrical stimulation. Results: ICP/MAP ratios in the RT group were significantly lower than those in the control group (p=0.009). In the RT group, only focal loss of hair on irradiated area was observed, and there were no significant physical and physiological changes. Conclusions: In SD rats, external pelvic radiation in a single 24 Gy fraction induced decreased erectile function significantly, and did not cause serious adverse events. These results suggest that a rat model induced by external pelvic radiation in a single 24 Gy fraction is feasible and safe.

MP86-03 THE INFLUENCE OF MACA (LEPIDIUM MEYENII) ON THE GLUCOSE METABOLISM AND ERECTILE FUNCTION IN TYPE 1 DIABETIC RATS

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology I1 Apr 2016MP86-03 THE INFLUENCE OF MACA (LEPIDIUM MEYENII) ON THE GLUCOSE METABOLISM AND ERECTILE FUNCTION IN TYPE 1 DIABETIC RATS Masaki Kimura, Amr Abdelhamed, Takahiro Noguchi, Takeshi Ashizawa, Hisashi Hirano, Hiroki Koyasu, Kazutaka Terai, Keisuke Saito, Hisamitsu Ide, Satoru Muto, Raizo Yamaguchi, and Shigeo Horie Masaki KimuraMasaki Kimura More articles by this author , Amr AbdelhamedAmr Abdelhamed More articles by this author , Takahiro NoguchiTakahiro Noguchi More articles by this author , Takeshi AshizawaTakeshi Ashizawa More articles by this author , Hisashi HiranoHisashi Hirano More articles by this author , Hiroki KoyasuHiroki Koyasu More articles by this author , Kazutaka TeraiKazutaka Terai More articles by this author , Keisuke SaitoKeisuke Saito More articles by this author , Hisamitsu IdeHisamitsu Ide More articles by this author , Satoru MutoSatoru Muto More articles by this author , Raizo YamaguchiRaizo Yamaguchi More articles by this author , and Shigeo HorieShigeo Horie More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2311AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Maca (Lepidium meyenii) is widely dispersed plant on high plateaus of the Andean mountain range in Peru. Maca is also known for its supportive effects on fertility and increase of libido. However, the influence of Maca on erectile dysfunction (ED) has not been fully investigated in diabetic rats. In the present study, we aim to investigate the influence of Maca on the Glucose metabolism and erectile function using streptozotocin (STZ)-induced type 1 diabetic rat. METHODS 40 adult male rats (8 weeks age) were divided into five groups, including control, STZ diabetic, STZ with Tadalafil, STZ with Maca, and STZ with Maca/Tadalafil. Diabetes was induced by a single intra-peritoneal injection of STZ in 0.1 citrate buffer (pH4.5) in a dose of 60mg/kg. Rats with blood glucose =200mg/dl were considered diabetic. STZ rats received daily Maca of 100mg/kg and/or Tadalafil of 1mg/kg for 8 weeks. Body weight and blood glucose level were measured at baseline and 8 weeks after treatment. Intracavernous pressure (ICP) measurements with cavernous nerve electrical stimulation were conducted after 8 weeks of treatment. We investigated the results of ICP/ mean arterial pressure (MAP) ratios for each rat. One way ANOVA test was used to test significance among the study groups. Each group contained 6 to 8 rats. RESULTS The body weight and blood glucose of STZ with Maca and STZ with Maca/Tadalafil groups were significantly decreased in comparison to STZ (P<0.001), and STZ with Tadalafil groups (P<0.001). In the STZ with Maca group, a significant increase in ICP/MAP was observed compared with STZ diabetic group (P<0.001). Similarly, In the STZ with Maca/Tadalafil group, a significant increase in ICP/MAP was observed compared with STZ diabetic group (P<0.001) and STZ with Tadalafil group (P=0.010). CONCLUSIONS Current data showed improvement of glucose metabolism and erectile function after 8 weeks of Maca treatment in STZ induced diabetic rats. Furthermore, synergetic effects of Maca along with PDE5 inhibitor administration were observed. Although more detailed mechanisms are needed to be elucidated, these findings suggest that Maca may exert beneficial effects for glucose tolerance and erectile dysfunction in diabetic patients. © 2016FiguresReferencesRelatedDetails Volume 195Issue 4SApril 2016Page: e1107 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Masaki Kimura More articles by this author Amr Abdelhamed More articles by this author Takahiro Noguchi More articles by this author Takeshi Ashizawa More articles by this author Hisashi Hirano More articles by this author Hiroki Koyasu More articles by this author Kazutaka Terai More articles by this author Keisuke Saito More articles by this author Hisamitsu Ide More articles by this author Satoru Muto More articles by this author Raizo Yamaguchi More articles by this author Shigeo Horie More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

Shengjing Capsule Improves Erectile Function Through Regulation of Nitric Oxide-induced Relaxation in Corpus Cavernosum Smooth Muscle in a Castrated Rat Model

To evaluate the effects of Shengjing capsule on erectile function in a castrated rat model, and further to investigate the underlying molecular mechanism. Thirty male Sprague-Dawley rats were randomly divided into six groups (n = 5 per group), including sham group, castration group, testosterone replacement group, high-dose Shengjing capsule group, medium-dose Shengjing capsule group, and low-dose Shengjing capsule group. The weight of the body and androgen-sensitive organs, and the serum level of testosterone were assessed. Erectile function was evaluated using cavernous nerve electrical stimulation after treatment. Corpus cavernosum tissue was examined by Masson's trichrome staining, immunohistochemistry, quantitative reverse transcriptase-polymerase chain reaction, and Western blot. Serum testosterone level, mean weights of body, and accessory sexual organs were not significantly different between Shengjing capsule treatment and the castration groups (P > .05 for all). Significant recovery of erectile function and the increased smooth muscle components were observed in the Shengjing capsule treatment group as compared with the castration group (P < .01 and P < .01, respectively). The gene and protein expression of 3 subtypes of nitric oxide synthase (NOS)-neuronal (nNOS), inducible (iNOS), and endothelial (eNOS) -in cavernous tissue in Shengjing capsule-treated rats were significantly higher than in the castration group (P < .05 for all). Phosphodiesterase type 5 messenger ribonucleic acid and protein expression in each group followed a trend similar to that of smooth muscle content. These results show that Shengjing capsule improves the erectile function by protecting the smooth muscle content and enhancing NOS activity in penile tissues of castrated male rats.