Cavernous Smooth Muscle Cells Research Articles

Exosomes secreted by adipose mesenchymal stem cells overexpressing circPIP5K1C exert

BackgroundErectile dysfunction (ED) seriously affects men's normal life, and obstructive sleep apnoea (OSA) has been diagnosed as a causative factor. Currently, exosomes secreted by adipose mesenchymal stem cells (ADSC) have been used in the non-clinical experimental treatment of ED disease with prominent efficacy due to the advantages of high stability and no immune exclusion. MethodsIn this study, chronic intermittent hypoxia (CIH) exposure was used to induce ED-corresponding phenotypes in Sprague Dawley (SD) rats as well as in cavernous smooth muscle cells (CCSMCs). ED symptoms were treated using exosomes secreted by ADSCs overexpressing circPIP5K1C (EXO-circ) injected into the rat corpus cavernosum. ResultsEXO-circ has the effect of ameliorating ED induced by CIH exposure in rats, the mechanism of which is to promote the expression of the downstream target gene SMURF1 after adsorption of miR-153-3p through the sponge so that SMURF1 and PFKFB3 occur protein-protein binding and ubiquitination degradation of PFKFB3 appears to inhibit the occurrence of spongiotic smooth muscle cells glycolysis, and to restore the function of the smooth muscle. ConclusionsThese findings show that EXO-circ have a promising therapeutic potential in OSA-induced ED.

Cldn4 overexpression promotes penile cavernous smooth muscle cell fibrotic response via the JNK signaling pathway.

Erectile dysfunction (ED), defined as the inability to achieve or maintain a penile erection sufficient to satisfy sexual behavior, is prevalent worldwide. Using previous research, bioinformatics, and experimental confirmation, we aimed to discover genes that contribute to ED through regulating hypoxia in corpus cavernosum smooth muscle cells (CCSMCs). We used the Gene Expression Omnibus to acquire the sequencing data of the corpus cavernosum transcriptome for diabetic ED and nerve injury type ED rats. We intersected the common differentially expressed genes. Further verification was performed using single cell sequencing. Real-time quantitative polymerase chain reaction and immunofluorescence were used to investigate whether the differentially expressed genes are found in the corpus cavernosum. We used induced hypoxia to assess cell viability changes, and we developed a lentivirus overexpressing Cldn4 for in vitro and in vivo experiments to measure changes in JNK signaling, fibrosis, hypoxia, and erectile function. Our results indicate that targeting the JNK pathway and decreasing local hypoxia may be better options for therapeutic intervention to improve erectile function. We identified Cldn4 and found its expression increased in the corpora cavernosa of the 2 datasets. In addition, we found that hypoxia can increase the expression of Cldn4, activate the JNK signaling pathway, and exacerbate fibrosis in CCSMCs. Cldn4 overexpression in CCSMCs activated the JNK signaling pathway and increased fibrotic protein expression. Last, rat corpus cavernosum overexpressing Cldn4 activated the JNK signaling pathway, increased local fibrosis, and impaired erectile function. Through bioinformatics and in vitro and in vivo experiments, we found that Cldn4 has a negative effect on ED, and targeting Cldn4 may provide new ideas for ED treatment. Although we have identified Cldn4 as a potential target for ED treatment, we have only conducted preliminary validation on CCMSCs, and we still need to further validate in other cell lines. CCSMC hypoxia leads to increased Cldn4, in both nerve injury and diabetic ED rat models, and promotes fibrosis by activating the JNK signaling pathway.

Simvastatin attenuates diabetes mellitus erectile dysfunction in rats by miR-9-5p-regulated PDCD4.

DMED is a common complication of diabetes, for which new treatment methods are urgently required. Focused on DMED, the pharmacological mechanism of simvastatin (Sim) was probed. A model of DMED was made in rats with streptozotocin and orally medicated with Sim. Lentiviral vectors that interfere with miR-9-5p or PDCD4 were injected, and the erectile function, histopathology of cavernous tissue, and α-SMA expression were evaluated. Cavernous smooth muscle cells (CMSCs) obtained from DMED rats were treated with Sim and transfected with the plasmid vector that interferes with miR-9-5p or PDCD4 to observe cell viability and apoptosis. The binding relationship between miR-9-5p and PDCD4 was checked. After 8-week treatment with Sim, erectile function was improved and the corpus cavernosum injury was alleviated. Upregulating miR-9-5p or downregulating PDCD4 further improved erectile function and cavernous injury in rats. miR-9-5p targeted regulation of PDCD4. In vitro cell experiment results showed that Sim induced proliferation and reduced apoptosis of CSMCs by enhancing miR-9-5p-targeted regulating PDCD4 in vitro. Sim attenuates DMED in rats via miR-9-5p/PDCD4.

Low androgen levels induce ferroptosis of rat penile cavernous endothelial cells.

Endothelial dysfunction caused by low androgen levels in penile tissue can lead to erectile dysfunction. The exact mechanism of endothelial dysfunction has not been thoroughly studied. The study sought to verify whether low androgen levels induce ferroptosis of endothelial cells in rat penile tissue. Rat penile cavernous endothelial cells (CP-R133) were divided into a no-androgen group (Dihydrotestosterone (DHT): 0nmol/L), very low-androgen group (DHT: 0.1nmol/L), low-androgen group (DHT: 1nmol/L), DHT = 10nmol/L group, DHT (0nmol/L) + ferrostatin-1 (Fer-1) group, DHT (0.1nmol/L) + Fer-1 group, DHT (1nmol/L) + Fer-1 group, DHT (10nmol/L) + Fer-1 group. Cell viability, intracellular ferrous ion (Fe2+), malondialdehyde (MDA), GSH into oxidized glutathione (GSSG), reactive oxygen species (ROS), nitric oxide (NO), transferrin receptor 1 protein (TfR1), solute carrier family 7 member 11 (SLC7A11), glutathione peroxidase 4 (GPX4), acyl-CoA synthetase long-chain family member 4 (ACSL4), endothelial nitric oxide synthase (eNOS), and phospho-eNOS (p-eNOS) were detected. Low androgen levels could induce ferroptosis of rat penile cavernous endothelial cells in vivo by upregulating the expressions of TfR1 and ACSL4 and downregulating the expressions of SLC7A11 and GPX4. Cell viability, the levels of glutathione (GSH), NO, SLC7A11, GPX4, and p-eNOS/eNOS in the DHT = 0nmol/L group were lower than those in the other groups (P < .05). The levels of Fe2+, ROS, MDA, GSSG, TfR1, and ACSL4 in the DHT = 0nmol/L group were higher than those in the other groups (P < .05). Cell viability and the levels of GSH, NO, SLC7A11, GPX4, and p-eNOS/eNOS in the DHT = 1nmol/L group were lower than those in the DHT (1nmol/L) + Fer-1 group, DHT = 10nmol/L group, and DHT (10nmol/L) + Fer-1 group (P < .05). The levels of Fe2+, ROS, MDA, GSSG, TfR1, and ACSL4 in the DHT = 1nmol/L group were higher than those in the DHT (1nmol/L) + Fer-1 group, DHT = 10nmol/L group, and DHT (10nmol/L) + Fer-1 group (P < .05). A ferroptosis inhibitor might be a novel drug for treating erectile dysfunction caused by low androgen level. The results of this study need to be further confirmed in in vitro and in human studies. Meanwhile, further investigation is needed to clarify whether low androgen levels affect ferroptosis of rat penile cavernous smooth muscle and nerve cells. Low androgen levels can induce ferroptosis of endothelial cells in rat penile tissue. Inhibition of ferroptosis can reverse endothelial dysfunction caused by low androgen levels.

Erectile Dysfunction: Key Role of Cavernous Smooth Muscle Cells.

Erectile dysfunction is increasingly affecting men, from the elderly to young adults, being a sexual disorder related to the inability to generate or maintain a penile erection. This disorder is related to psychosocial factors such as anxiety, depression, and low self-esteem, to organic factors such as the presence of preexisting conditions like hypertension, diabetes and dyslipidemia. The pathophysiology of the disease is related to changes in the neurotransmission of the autonomic or the non-cholinergic non-adrenergic nervous system, as well as the release of local mediators, such as thromboxane A2 and endothelin, and hormonal action. These changes lead to impaired relaxation of cavernous smooth muscle, which reduces local blood flow and impairs penile erection. Currently, therapy is based on oral vasodilation, such as sildenafil, tadalafil, vardenafil and iodenafil, or by direct administration of these agents into the corpus cavernosum or by intraurethral route, such as alprostadil and papaverine. Despite this, studies that consolidate the understanding of its pathophysiological process contribute to the discovery of new more efficient drugs for the treatment of erectile dysfunction. In this sense, in the present work an extensive survey was carried out of the mechanisms already consolidated and the most recent ones related to the development of erectile dysfunction.

MP38-08 HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS AMELIORATE ERECTILE DYSFUNCTION IN TYPE I AND TYPE II DIABETES MELLITUS RATS THROUGH THE ATTENUATION OF FERROPTOSIS BY SLC7A11/GPX4/ACSL4 PATHWAY

You have accessJournal of UrologyCME1 May 2022MP38-08 HUMAN UMBILICAL CORD MESENCHYMAL STEM CELLS AMELIORATE ERECTILE DYSFUNCTION IN TYPE I AND TYPE II DIABETES MELLITUS RATS THROUGH THE ATTENUATION OF FERROPTOSIS BY SLC7A11/GPX4/ACSL4 PATHWAY Huan Feng, Tao Wang, and Jihong Liu Huan FengHuan Feng 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.0000000000002592.08AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Diabetic Erectile Dysfunction (DMED) is a common complication of diabetes, which seriously impairs the life of male patients of all ages. Diabetes could destroy various cells in the corpus cavernosum, such as nerve cells, endothelial cells, smooth muscle cells. Human Umbilical Cord Mesenchymal Stem Cells (hUCMSCs) are cells with multipotential differentiation potential.The present study aims to investigate the therapeutic role and mechanism of hUCMSCs on type I and type II diabetes mellitus rats. METHODS: Fifty male Sprague-Dawley rats received intraperitoneal injection with streptozotocin for 3 days to induce type 1 diabetes. Another 50 male ZDF rats were fed with Purina 5008 to induce type 2 DMED. After 8 weeks, type 1 and type 2 DMED rats were separately selected by the apomorphine (APO) test. Both of types DMED rats were divided into normal control group (NC), diabetic group (DM), cavernous injection (1×106) group (CI), tail vein injection (1×106) group (VI). Stem cells were labeled with Edu prior to injection for subsequent observation. Primary corpus cavernous smooth muscle cells (CCSMCs) from healthy rats were cultured and treated with berberine. Mitochondria in ccSMCs were observed by transmission electron microscopy (TEM). RESULTS: None of rats received stem cell injection had obvious adverse reactions. Injection of hUCMSCs improved erectile function, reduced levels of NADPH oxidases, ROS production and tissue iron content. Among the type 1 DMED rats, the results showed that mICP/MAP of DM (0.232±0.082 mmHg) was significantly lower than NC (0.793±0.153 mmHg), CI (0.333±0.104 mmHg) and VI (0.384±0.095 mmHg) were improved compared with DM, and similar phenomenon also showed among the type 2 DMED rats. Besides, co-cultured CCSMCs with hUCMSCs exhibited a lower level of ferroptosis by SLC7A11/GPX4/ACSL4 pathway with high glucose (HG) treatment. TEM showed that the mitochondria shrunk, the cristae of mitochondria were destroyed, and exhibited high electron density in CCSMCs treated with HG, and these effects could be obviously reversed by hUCMSCs. CONCLUSIONS: Human umbilical cord mesenchymal stem cells can safely and effectively ameliorate erectile dysfunction in both type I and type II diabetes mellitus rats, and attenuate ferroptosis of CCSMCs through SLC7A11/GPX4/ACSL4 pathway. It provides preclinical evidence for application of hUCMSCs to DMED treatment and potential molecular mechanism of it. Source of Funding: None © 2022 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 207Issue Supplement 5May 2022Page: e621 Advertisement Copyright & Permissions© 2022 by American Urological Association Education and Research, Inc.MetricsAuthor Information Huan Feng More articles by this author Tao Wang More articles by this author Jihong Liu More articles by this author Expand All Advertisement PDF DownloadLoading ...

Retraction Statement.

Retraction Statement.

Berberine ameliorates erectile dysfunction in rats with streptozotocin-induced diabetes mellitus through the attenuation of apoptosis by inhibiting the SPHK1/S1P/S1PR2 and MAPK pathways.

The population with diabetes mellitus-induced erectile dysfunction is increasing rapidly, but current drugs are not effective in treating erectile dysfunction. Studies of the traditional Chinese medicine extract berberine on diabetes and its complications provide us with new ideas. To evaluate the therapeutic effect and potential mechanism of berberine on the erectile function of diabetic rats. Fifty male Sprague-Dawley rats were randomly grouped, and 42 rats were injected intraperitoneally with streptozotocin to establish a diabetes model. Erectile dysfunction rats were screened out through the apomorphine test and randomly divided into the diabetes mellitus and berberine groups, and these animals were administered berberine (200mg/kg/day) and normal saline by gavage for 4weeks. Primary corpus cavernous smooth muscle cells from healthy rats were cultured and treated with berberine. Fasting blood glucose in the diabetes mellitus group was significantly increased, while berberine showed no significant effect on glucose. Erectile function was obviously impaired in the diabetes mellitus group, and berberine administration partially rescued this impairment. The expression of sphingosine kinase 1, S1PR2, and sphingosine-1-phosphate in the diabetes mellitus group was increased. Berberine partially inhibited the expression of sphingosine kinase 1 and S1PR2, but the decrease in sphingosine-1-phosphate was not significant. Moreover, mitogen-activated protein kinase pathway factor expression was upregulated and eNOS activity was decreased in the diabetes mellitus group. Berberine treatment could partially reverse these alterations. Severe fibrosis and apoptosis were detected in diabetic rats, accompanied by higher expression of TGFβ1, collagen I/IV, Bax/Bcl-2, and caspase 3 than in the other groups. However, supplementation with berberine inhibited the expression of these proteins and attenuated fibrosis and apoptosis. Berberine ameliorated erectile dysfunction in rats with diabetes mellitus, possibly by improving endothelial function and inhibiting apoptosis and fibrosis by suppressing the sphingosine kinase 1/sphingosine-1-phosphate/S1PR2 and mitogen-activated protein kinase pathways.

Improvement of damaged cavernosa followed by neuron-like differentiation at injured cavernous nerve after transplantation of stem cells seeded on the PLA nanofiber in rats with cavernous nerve injury.

This study investigated the differentiation of transplanted transplanted mesenchymal stem cells MSCs into neuron-like cells, repair of erectile dysfunction (ED), and synergy of MSCs seeded to nanofibrous scaffolds with after transplantation around the injured cavernous nerve (CN) of rats. The synthesized polymer was electrospun in a rotating drum to prepare nanofiber meshes (NMs). Human MSCs were prepared and confirmed. Eight-week-old male Sprague-Dawley rats were divided into five groups of six each: group 1-sham operation; group 2-CN injury; group 3-MSCs treatment after CN injury; group 4-nanofibrous scaffold treatment after CN injury; and group 5-post-CN injury treatment combining a nanofibrous scaffold and MSCs (nano-MSCs). In the latter group, the damaged CN was instantly surrounded by an MSC-containing a nanofibrous scaffold in the aftermath of injury. Morphological analysis and immuno-histochemical staining in relation to nerves (Tuj1, NF, MAP2, MBP and peripherin), endothelium (vWF), smooth muscle (SMA), neurofilament (NF), and apoptosis (TUNEL) were performed. We evaluated the mean proportion expressed as a percentage of the ratio of muscle to collagen of penile cavernous smooth-muscle cells as well as the expression of cavernous SMA, NF, vWF, and TUNEL makers. Compared to the group free of CN injury, erectile function was markedly reduced in the group with CN injury at 2 and 4weeks (p < 0.05). By contrast, compared to the sham operation group, erectile function was better in the group with MSC transplantation (p < 0.05). Similarly, by comparison to the group solely with hMSCs, erectile function was better in the group with nano-MSC transplantation (p < 0.05). Transplantation of MSCs demonstrated the neuronal differentiation. By contrast to MSCs on their own, neuronal differentiation was more significantly expressed in nano-MSCs. The mean proportion expressed as a percentage of the ratio of muscle to collagen of penile cavernous smooth-muscle cells, the expression of cavernous SMA, NF, vWF, and apoptosis improved in the cavernosum after transplantation. NMs showed synergy with MSCs for the repair of erectile dysfunction. Transplanted MSCs differentiated into neuron-like cells and repaired erectile dysfunction in the rats with CN injury. Transplanted MSCs increased the mean percentage of the collagen area of the caversnosum as well as the expression levels of cavernous neuronal, endothelial, smooth-muscle markers, and apoptosis.

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.

Ionic Channels as Potential Therapeutic Targets for Erectile Dysfunction: A Review.

Erectile dysfunction (ED) is a prevalent condition, especially in men over 40 years old, characterized by the inability to obtain and/or maintain penile erection sufficient for satisfactory sexual intercourse. Several psychological and/or organic factors are involved in the etiopathogenesis of ED. In this context, we gathered evidence of the involvement of Large-conductance, Ca2+-activated K+ channels (BKCa), Small-conductance, Ca2+-activated K+ channels (SKCa), KCNQ-encoded voltage-dependent K+ channels (KV7), Transient Receptor Potential channels (TRP), and Calcium-activated Chloride channels (CaCC) dysfunctions on ED. In addition, the use of modulating agents of these channels are involved in relaxation of the cavernous smooth muscle cell and, consequent penile erection, suggesting that these channels are promising therapeutic targets for the treatment of erectile dysfunction.

Human Tissue Kallikrein 1 Improves Erectile Dysfunction of Streptozotocin-Induced Diabetic Rats by Inhibition of Excessive Oxidative Stress and Activation of the PI3K/AKT/eNOS Pathway

Objective To investigate the protective effects and mechanisms of human tissue kallikrein 1 (hKLK1) on type 1 diabetes mellitus- (DM-) induced erectile dysfunction in rats. Materials and Methods. The homozygous transgenic rats (TGR) harboring the hKLK1 gene and age-matched wild-type Sprague Dawley rats (WTR) were involved, and intraperitoneal injection of streptozotocin was utilized to induce diabetes in rats. Forty-eight-week-old male rats were randomly divided into a WTR group, TGR group, diabetic WTR group (WTDM), diabetic TGR group (TGDM), and TGDM with HOE140 group (TGDMH), with eight rats in each group. Twelve weeks later, the erectile response of all rats was detected by cavernous nerve electric stimulation, and corpus cavernosums were harvested to evaluate the levels of cavernous oxidative stress (OS), apoptosis, fibrosis, and involved pathways. Moreover, cavernous smooth muscle cells (CSMC) and endothelial cells (EC) were primarily isolated to build a coculture system for a series of in vitro verification. Results The hKLK1 gene and age-matched wild-type Sprague Dawley rats (WTR) were involved, and intraperitoneal injection of streptozotocin was utilized to induce diabetes in rats. Forty-eight-week-old male rats were randomly divided into a WTR group, TGR group, diabetic WTR group (WTDM), diabetic TGR group (TGDM), and TGDM with HOE140 group (TGDMH), with eight rats in each group. Twelve weeks later, the erectile response of all rats was detected by cavernous nerve electric stimulation, and corpus cavernosums were harvested to evaluate the levels of cavernous oxidative stress (OS), apoptosis, fibrosis, and involved pathways. Moreover, cavernous smooth muscle cells (CSMC) and endothelial cells (EC) were primarily isolated to build a coculture system for a series of Conclusions hKLK1 preserves erectile function of DM rats through its antitissue excessive OS, apoptosis, and fibrosis effects, as well as activation of the PI3K/AKT/eNOS/cGMP pathway in the penis. Moreover, hKLK1 promotes relaxation and prevents high glucose-induced injuries of CSMC mediated by EC-CSMC crosstalk.

Simvastatin alleviated diabetes mellitus-induced erectile dysfunction in rats by enhancing AMPK pathway-induced autophagy.

Diabetes mellitus-induced erectile dysfunction is a common diabetic complication, and new therapeutics and the pathogenesis of diabetes mellitus-induced erectile dysfunction need to be investigated. The aim was to investigate the pathogenesis of diabetes mellitus-induced erectile dysfunction and the pharmacological mechanism of simvastatin treatment in diabetes mellitus-induced erectile dysfunction model rats. A total of 86 male Sprague Dawley rats aged 8weeks old were used in this study. The rats were divided into three groups: control (normal), diabetes mellitus-induced erectile dysfunction (streptozotocin-injected), and diabetes mellitus-induced erectile dysfunction+simvastatin (sim). Each group was subdivided into two subgroups for in vitro and in vivo analyses. A bioinformatics method was used to detect differences in gene expression in the corpus cavernosum between normal and diabetes mellitus-induced erectile dysfunction rats. Erectile function was measured by a cavernous nerve electrostimulation test. Corpus cavernosum fibrosis was assessed by Masson staining and Western blotting. Immunofluorescence and Western blotting were performed to explore the differential expression of autophagy-related genes and the AMPK-SKP2-CARM1 pathway genes in rat cavernous smooth muscle cells and the corpus cavernosum. The autophagosomes of the corpus cavernosum tissue were observed by transmission electron microscopy. Autophagy-related genes and pathways (the AMPK and FoxO pathway) were identified by bioinformatics analysis and confirmed at the protein level. Simvastatin, an AMPK agonist, was used to treat diabetes mellitus-induced erectile dysfunction rats for 8weeks, demonstrating that erectile function was improved for 80.5% (P<.05) of rats. Corpus cavernosum fibrosis was alleviated (P<.05), and autophagy was further enhanced (P<.05); these results might be partially caused by AMPK-SKP2-CARM1 pathway activation (P<.05). Simvastatin could enhance protective autophagy by activating the AMPK-SKP2-CARM1 pathway to improve erectile function in diabetes mellitus-induced erectile dysfunction rats.

MicroRNA-205 is associated with diabetes mellitus-induced erectile dysfunction via down-regulating the androgen receptor.

As a major class of regulatory genes in majority metazoans, microRNAs (miRs) play an important role in various diseases including diabetes mellitus (DM). Lack of androgens has previously been associated with DM‐induced erectile dysfunction (DMED). In addition, the biological functioning of androgen is mediated by androgen receptor (AR). Herein, we sought to investigate whether miRs participate in AR‐associated DMED. Sprague‐Dawlay rats were employed to establish DMED models. After modelling, levels of miR‐205 and AR in their cavernous bodies were measured. The relationship between miR‐205 and AR was verified using a dual‐luciferase reporter gene assay. The underlying regulatory mechanisms of miR‐205 were investigated in concert with the treatment of mimics or inhibitors of miR‐205, or AR overexpression in the cavernous smooth muscle cells (CSMCs) isolated from rats with DMED. Meanwhile, the effects of miR‐205 and AR on cell proliferation and apoptosis were evaluated using MTT assay and flow cytometry respectively. Rats with DMED presented with increased miR‐205 and decreased AR levels in the cavernous bodies. AR was identified as a target gene of miR‐205. Down‐regulation of miR‐205 or up‐regulation of AR could increase proliferation and inhibits apoptosis of CSMCs in addition to improvements in the erectile functioning of rats with DMED. In summary, miR‐205 may contribute to the pathogenesis of DMED via down‐regulation of AR expressions.

Testosterone increases corpus cavernous smooth muscle cells in oxidative stress-induced rodents (Sprague-Dawley)

Testosterone increases corpus cavernous smooth muscle cells in oxidative stress-induced rodents (Sprague-Dawley)

161 Low Intensity Extracorporeal Shock Wave Therapy Activates Endogenous Progenitor/Stem Cells as Indicated by Histone 3 Phosphorylation

Low Intensity Extracorporeal Shock Wave Therapy (Li-ESWT) has been employed for many years to treat ED yet its mechanism of action remains poorly defined. We have previously demonstrated that the therapeutic effects from Li-ESWT may be partially related to the activation of endogenous progenitor/stem cells and we are endeavoring to further describe these pathways. To determine the mechanism by which Li-ESWT activates endogenous stem cells by interrogating histone 3 phosphorylation, which occurs only during the mitotic phase of the cell cycle. Male Sprague-Dawley (SD) rats were treated with 5-ethynyl-2'-deoxyuridine (EdU) pulse followed by Li-ESWT, and cohort analysis was performed. Additionally, rat adipose derived stem cells (RADSCs) and rat penile cavernous smooth muscle cells (RCSMCs) were also treated with Li-ESWT. Fifty male SD rats at 12 weeks old were grouped into three cohorts: 1). Sham; 2). Sub-acute: the animals underwent bilateral cavernous nerve injury (BCNI) then were separated into control and Li-ESWT groups. One week after BCNI, the Li-ESWT group were injected with EdU (50mg/kg, i.p) 30 minutes before Li-ESWT (0.02mJ/mm2, 3 Hz, 300 pulses). One week later, ICP/MAP were measured. 3). Chronic: the animals underwent BCNI then grouped into control and Li-ESWT groups. One week after BCNI, the animals were treated with Li-ESWT (0.02mJ/mm2, 3 Hz, 300 pulses) twice a week for 4 weeks. EdU was injected 30 min before the first and last Li-ESWT. One week after the last Li-ESWT, ICP/MAP were measured. The penile erectile tissues and major pelvic ganglion (MPG) were harvested for histological study to assess smooth muscle/collagen content and endothelium content in the corpora cavernosa. EdU as marker of cell activation and phosphorylated histone 3 (H3P) as marker of the mitotic phase of the cell cycle were also assessed. To confirm the effect of Li-ESWT on the phosphorylation of H3 in vitro, RADSCs and RCSMCs were treated with Li-ESWT (0.02mJ/mm2, 3 Hz, 100 pulses), and EdU incorporation rate and the expression of H3P was assessed.

MicroRNA-141 Inhibits the Proliferation of Penile Cavernous Smooth Muscle Cells Associated with Down-Regulation of the Rhoa/Rho Kinase Signaling Pathway

The role of the RhoA/Rho kinase signaling pathway in diabetes mellitus-induced erectile dysfunction has been partially understood. In the present study, we explored the changes of the RhoA/Rho associated kinase (ROCK) signaling pathway in diabetic erectile dysfunction in vivo and the effects of microRNA-141 on the RhoA/ROCK signaling pathway in vitro. The mRNA and protein expressions of RhoA and ROCK2 were significantly increased while the expression of microRNA-141 was decreased in the penile cavernous smooth muscle cells of rats with diabetic erectile dysfunction. Moreover, increased expression of microRNA-141, decreased expressions of RhoA and ROCK2 (mRNA and protein), accelerated cell proliferation rate and reduced cell apoptosis were found in the microRNA-141 mimics group and the siRNA-Rho group. The microRNA-141 expression in the microRNA-141 inhibitors + siRNA-Rho group was significantly decreased. microRNA-141 specifically bound to Rho-3'-UTR and down-regulated the expression of Rho gene at the post transcriptional level. Decreased expression of miR-141 is associated with up-regulation of RhoA and ROCK2 in the RhoA/ROCK signaling pathway in rats with diabetic erectile dysfunction. miR-141 inhibits the growth of penile cavernous smooth muscle cells associated with down-regulation of the RhoA/ROCK signaling pathway in vitro.

Additive effects of the Rho kinase inhibitor Y-27632 and vardenafil on relaxation of the corpus cavernosum tissue of patients with erectile dysfunction and clinical phosphodiesterase type 5 inhibitor failure.

To evaluate the expression of the Rho/Rho-associated protein kinase (ROCK) pathway in the corpus cavernosum of patients with severe erectile dysfunction (ED) compared with healthy human corpus cavernosum, and to test the functional effects of two Rho kinase inhibitors (RKIs) on erectile tissue of patients with severe ED, which did not respond to phosphodiesterase type 5 inhibitors (PDE5Is). Human corpus cavernosum samples were obtained after consent from men undergoing penile prosthesis implantation (n = 7 for organ bath experiments, n = 17 for quantitative PCR [qPCR]). Potent control subjects (n = 5) underwent penile needle biopsy. qPCR was performed for the expression of RhoA and ROCK subtypes 1 and 2. Immunohistochemistry staining against ROCK and α smooth muscle actin (αSMA) was performed on the corpus cavernosum of patients with ED. Tissue strips were precontracted with phenylephrine and incubated with 1 μm of the PDE5I vardenafil or with DMSO (control). Subsequently, increasing concentrations of the RKIs azaindole or Y-27632 were added, and relaxation of tissue was quantified. The expression of ROCK1 was unchanged (P > 0.05), while ROCK2 (P < 0.05) was significantly upregulated in patients with ED compared with controls. ROCK1 and ROCK2 protein colocalized with αSMA, confirming the presence of this kinase in cavernous smooth muscle cells and/or myofibroblasts. After incubation with DMSO, 10 μm azaindole and 10 μm Y-27632 relaxed precontracted tissues with 49.5 ± 7.42% (P = 0.1470 when compared with vehicle) and 85.9 ± 10.3% (P = 0.0016 when compared with vehicle), respectively. Additive effects on relaxation of human corpus cavernosum were seen after preincubation with 1 μm vardenafil. The RKI Y-27632 causes a significant relaxation of corpus cavernosum in tissue strips of patients with severe ED. The additive effect of vardenafil and Y-27632 shows that a combined inhibition of Rho-kinase and phosphodiesterase type 5 could be a promising orally administered treatment for severe ED.

MP86-12 HUMAN TISSUE KALLIKREIN 1 RESTORES ERECTILE FUNCTION OF STREPTOZOTOCIN- INDUCED DIABETIC RATS BY ACTIVATING PI3K/ENOS PATHWAY, INHIBITING OXIDATIVE STRESS AND APOPTOSIS IN THE CORPUS CAVERNOSUM

You have accessJournal of UrologySexual Function/Dysfunction: Basic Research & Pathophysiology I1 Apr 2016MP86-12 HUMAN TISSUE KALLIKREIN 1 RESTORES ERECTILE FUNCTION OF STREPTOZOTOCIN- INDUCED DIABETIC RATS BY ACTIVATING PI3K/ENOS PATHWAY, INHIBITING OXIDATIVE STRESS AND APOPTOSIS IN THE CORPUS CAVERNOSUM Yang Luan, Yajun Ruan, Tao Wang, Yan Zhang, Kai Cui, Rui Li, Jun Yang, Ruibao Chen, Shaogang Wang, Jihong Liu, and Zhangqun Ye Yang LuanYang Luan More articles by this author , Yajun RuanYajun Ruan More articles by this author , Tao WangTao Wang More articles by this author , Yan ZhangYan Zhang More articles by this author , Kai CuiKai Cui More articles by this author , Rui LiRui Li More articles by this author , Jun YangJun Yang More articles by this author , Ruibao ChenRuibao Chen More articles by this author , Shaogang WangShaogang Wang More articles by this author , Jihong LiuJihong Liu More articles by this author , and Zhangqun YeZhangqun Ye More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2016.02.2320AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Our previous study has proved the protective role of human tissue kallikrein 1 (hKLK1) in erectile function of aged rats with the transgenic Sprague-Dawley (SD) rats harboring hKLK1 gene (TGR), but its effect on diabetic erectile dysfunction (ED) is still unknown. METHODS Eight-week-old male wild type SD rats (WTR) and age matched TGR were randomly divided into five groups: citrate buffer treated (1) WTR and (2) TGR; streptozotocin-induced (3) diabetic wild type rats (WTDM) and (4) diabetic transgenic rats (TGDM); (5) TGDM with daily administration of bradykinin receptor 2 inhibitor HOE 140 (TGDMH). Erectile response was measured by cavernous nerve electrostimulation after 12-week treatment. Then the corpus cavernosum (CC) tissues were harvested for hKLK1 gene identification and mechanism exploration. The regulation of hKLK1 on PI 3 kinase (PI3K)/ endothelial nitric oxide synthase (eNOS) pathway was detected within CC in vivo and TGR derived cavernous smooth muscle cell (CSMC) in vitro. In addition, the effect of hKLK1 on oxidative stress and apoptosis in CC of diabetic rats and high glucose treated CSMC was also evaluated. RESULTS hKLK1 attenuated the decreased erectile function of WTDM, while the protective effect was abolished in TGDMH. The decrease of PI3K(p55), AKT, eNOS, phospho-eNOS(Ser1177), cyclic guanosine monophosphate (cGMP) and increase of phospho-eNOS(Thr495), phosphodiesterase type 5, L-type calcium channel in CC of WTDM were improved in that of TGDM, but deteriorated again by HOE 140. Meanwhile, hKLK1 could elevate the level of cGMP and reduce the concentration of Ca2+ in CSMC in vitro, but these changes were diminished by B2R, PI3K or eNOS inhibitors. In addition, the CC of WTDM had higher proteins expression of nicotinamide adenine dinucleotide phosphate oxidase, malondialdehyde level, lower activity of superoxide dismutase and elevated apoptosis percentage when compared to WTR. However, hKLK1 improved these parameters in TGDM and inhibition with HOE 140 would abolish these advantages. Besides, the level of reactive oxygen species (ROS) and apoptosis rate elevated in high glucose treated CSMC in a concentration-dependent manner and hKLK1 could considerably inhibit the generation of ROS and cell apoptosis in vitro. CONCLUSIONS These results indicate that hKLK1 could restore diabetic ED through activation of PI3K/eNOS signaling and inhibition of oxidative stress and apoptosis in CC. © 2016FiguresReferencesRelatedDetailsCited bySeftel A (2016) Re: Preserved Erectile Function in the Aged Transgenic Rat Harboring Human Tissue Kallikrein 1Journal of Urology, VOL. 197, NO. 2, (493-493), Online publication date: 1-Feb-2017. Volume 195Issue 4SApril 2016Page: e1110 Advertisement Copyright & Permissions© 2016MetricsAuthor Information Yang Luan More articles by this author Yajun Ruan More articles by this author Tao Wang More articles by this author Yan Zhang More articles by this author Kai Cui More articles by this author Rui Li More articles by this author Jun Yang More articles by this author Ruibao Chen More articles by this author Shaogang Wang More articles by this author Jihong Liu More articles by this author Zhangqun Ye More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...

AB106. Role of Schwann cells in the regeneration of penile and peripheral nerves

Schwann cells (SCs) are the principal glia of the peripheral nervous system. The end point of SC development is the formation of myelinating and non-myelinating cells which ensheath large and small diameter axons, respectively. They play an important role in axon regeneration after injury, including cavernous nerve injury that leads to erectile dysfunction (ED). Although surgical techniques for radical prostatectomy (RP) for the treatment of prostate cancer have improved with the introduction of nerve-sparing techniques and robotic procedures, many patients still suffer from ED following surgery. During surgery trauma causes traction injury and local inflammatory changes in the neuronal microenvironment of the autonomic fibers innervating the penis resulting pathophysiological alterations in the end organ. The suggested pathophysiologic mechanisms responsible for RP-induced ED include: reduction in neuronal nitric oxide synthase (nNOS), a decrease in the number of cavernous endothelial and smooth muscle cells, increase in reactive oxygen species, and up-regulation of profibrotic factors and cavernous fibrosis as a result of cavernous hypoxia. Current research strategies to prevent ED following RP have focused on pharmacological interventions, such as PDE5 inhibitors, that prevent penile corporal fibrosis in an effort to preserve the hemodynamic mechanisms of penile erection. However, a number of patients with RP-induced ED do not respond well to such pharmacological intervention suggesting a need for novel therapies. Currently there are no interventions targeted at cavernosal nerve regeneration because the mechanisms of regeneration including demyelination, axon degeneration, re-myelination, SC and neuron interaction, the molecular signaling process is poorly understood. For decades SCs received relatively little scientific attention with poor understanding of their function despite the fact that they outnumber the neurons in the peripheral nerve system (PNS) and are involved in almost all neural functions. New technologies available to neurobiologists have now provided unexpected insight into SC function and have greatly expanded research.