Diabetes Mellitus Erectile Dysfunction Research Articles

Inhibition of YAP1 Rescues Erectile Dysfunction by Inhibiting Phenotypic Modulation through Myocardin in Diabetic Rats

Background. Phenotypic modulation was shown to be related to the pathological mechanism of diabetes mellitus erectile dysfunction (DMED), and excessive expression of YAP1 was measured in the cavernous tissues of rats with DMED. Objectives. This investigation was performed to explore the efficiency and mechanism of gene therapy through the knockdown of YAP1 in the field of ED treatment. Materials and Methods. Corpus cavernosum smooth muscle cells were obtained and cultivated in vitro employing the explant technique. EdU, CCK-8, and collagen gel lattice contraction assays were subsequently performed to assess cell proliferation and contractility, which indicate smooth muscle phenotypes. Results. Initially, overexpression of YAP1 enhanced the proliferation and expression of YAP1 and osteopontin (OPN) while reducing cell contractility and the expression of myocardin, α-SMA, and calpontin, which is consistent with phenotypic modulation. Moreover, knocking down YAP1 resulted in the opposite effects, and the blockade of myocardin attenuated the reversal of phenotypic transformation. Then, 45 screened DMED rats were randomly divided into three equal groups, named streptozotocin (STZ)-induced DMED with intracavernosal injection of Ad-myocardin (DMED + myocardin), DMED with lenti-shYAP1 (DMED + shYAP1) and DMED with PBS (DMED + PBS), and 10 rats were in the negative control group. Staining with H&E and Masson’s trichrome, immunochemistry, and determination of the ratio of the intracavernosal pressure/mean arterial pressure were performed to evaluate penile erection and histological changes. Western blotting was conducted to determine the molecular protein expression levels. In vivo, similar to myocardin overexpression, YAP1 inhibition rescued erectile function, repaired morphology, and maintained the contractile phenotype of smooth muscle tissues 14 days after treatment. Discussion and Conclusion. In conclusion, our study demonstrated the validity of gene therapy by downregulating excessive YAP1 expression to ameliorate ED in diabetic rats, and the ability of YAP1 inhibition to reverse phenotypic modulation was mediated through interaction with myocardin.

The mechanistic study on Wa medicine Niang-Mu-Liang medicinal liquor mitigating diabetes mellitus erectile dysfunction in rats by inhibiting ferroptosis

The mechanistic study on Wa medicine Niang-Mu-Liang medicinal liquor mitigating diabetes mellitus erectile dysfunction in rats by inhibiting ferroptosis

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.

Combination of mesenchymal stem cells carried on artificial extracellular matrix with LIPUS is a potential therapy that may cure diabetes mellitus erectile dysfunction

Diabetes mellitus erectile dysfunction (DMED) is a common chronic disease in andrology, which often responds insufficiently to routine treatment. Mesenchymal stem cell (MSC) treatment, as an important regenerative therapy, has the potential to reverse pathological changes and cure ED, but it is not satisfactory due to the limitation of the survival rate and retention rate. Meanwhile, the artificial extracellular matrix (ECM) shows potent characteristics of enhancing the biological behavior and activity of stem cells. Low intensity pulsed ultrasound (LIPUS) has also been widely used in regenerative medicine, which can promote cell proliferation and tissue repair. LIPUS can also stimulate the paracrine function of MSCs. These technologies have their own unique mechanisms and are of great potentiality in stem cell research. We hypothesized that MSCs carried on artificial ECM combined with LIPUS may play a synergistic role, which further improved the curative effect on DMED. This concept of combined therapy may be a prospective direction of regenerative therapy to cure DMED.

Combining transcriptomic analysis and network pharmacology to explore the mechanism by which Shaofu Zhuyu decoction improves diabetes mellitus erectile dysfunction

BackgroundErectile dysfunction is common among the complications of diabetes mellitus. Shaofu Zhuyu decoction (SFZYD) is commonly used to treat diabetic mellitus erectile dysfunction (DMED). However, its main active components and specific mechanism are still unknown. PurposeTo confirm the activity of SFZYD in improving DMED, explore the main active components of SFZYD, and clarify the underlying mechanism. MethodsA diabetic rat model was induced with streptozotocin (STZ). After intragastric administration, erectile function was assessed by the maximum intracavernous pressure (ICPmax)/mean arterial pressure (MAP). Corpus cavernosum fibrosis was evaluated by Masson staining, and ELISA methods were used to determine the serum levels of IL-6, TNF-α, IL-10, IL-4 and IL-1β to evaluate inflammation. Then, the main active components of SFZYD were identified by UPLC‒MS/MS. Finally, the target and biological mechanism of SFZYD in improving DMED were predicted by combined network pharmacology and transcriptomics, which was also validated by molecular docking and cellular thermal shift assay (CETSA) experiments. ResultsSFZYD significantly improved erectile dysfunction and inhibited inflammatory responses and local tissue fibrosis in diabetic rats. A total of 1846 active components were identified by UPLC‒MS/MS, and isorhamnetin was the main active component. The transcriptomic results were used to identify differentially expressed genes among the control, DM and SFZYD groups, and 1264 differentially expressed genes were obtained from the intersection. The network pharmacology results showed that SFZYD acts on core targets such as AKT1, ALB, HSP90AA1 and ESR1 through core components such as isorhamnetin, quercetin and chrysophanic acid. Further combined analysis revealed that multiple targets, such as CYP1B1, DPP4, NOS2 and LCN2, as well as the regulation of the PI3K-AKT signaling pathway, may be important mechanisms by which SFZYD improves DMED. Molecular docking verification showed that isorhamnetin, the key component of SFZYD, has good binding ability with several core targets, and its binding ability with CYP1B1 was the strongest. The CETSA results showed that isorhamnetin binds to CYP1B1 in CCECs. ConclusionSFZYD improves DMED, inhibits the inflammatory response and alleviates local tissue fibrosis. The combined application of transcriptomic, network pharmacology, molecular docking and CETSA approaches was helpful for revealing the mechanism by which SFZYD improves DMED, which may be related to the regulation of CYP1B1 and the PI3K-Akt signaling pathway.

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.

Mechanism of the Xuefu Zhuyu decoction in treating diabetes mellitus erectile dysfunction based on the CaSR/Gq-PLC-PKC pathway

Mechanism of the Xuefu Zhuyu decoction in treating diabetes mellitus erectile dysfunction based on the CaSR/Gq-PLC-PKC pathway

N‐acetylcysteine improves diabetic associated erectile dysfunction in streptozotocin‐induced diabetic mice by inhibiting oxidative stress

Oxidative stress appears to play a role in the pathogenesis of diabetes mellitus erectile dysfunction (DMED). This study aimed to investigate the effect of N‐acetylcysteine (NAC) on DMED in streptozotocin‐induced diabetic mice and to explore potential mechanisms. In the present study, we show that an erectile dysfunction is present in the streptozotocin‐induced mouse model of diabetes as indicated by decreases in intracavernous pressure responses to electro‐stimulation as well as from results of the apomorphine test of erectile function. After treatment of NAC, the intracavernous pressure was increased. In these DMED mice, oxidative stress and inflammatory responses were significantly reduced within the cavernous microenvironment, while activity of antioxidant enzymes in this cavernous tissue was enhanced after NAC treatment. These changes protected mitochondrial stress damage and a significant decreased in apoptosis within the cavernous tissue of DMED mice. This appears to involve activation of the nuclear factor erythroid 2‐like‐2 (Nrf2) signalling pathway, as well as suppression of the mitogen‐activated protein kinase (MAPK) p38/ NF‐κB pathway within cavernous tissue. In conclusion, NAC can improve erectile function through inhibiting oxidative stress via activating Nrf2 pathways and reducing apoptosis in streptozotocin‐induced diabetic mice. NAC might provide a promising therapeutic strategy for individuals with DMED.

Identification of Key microRNAs in Diabetes Mellitus Erectile Dysfunction Rats with Stem Cell Therapy by Bioinformatic Analysis of Deep Sequencing Data.

PurposeDiabetes mellitus erectile dysfunction (DMED) is a common resulting complication of diabetes. Studies have shown mesenchymal stem cell (MSC)-based therapy was beneficial in alleviating erectile function of DMED rats. While the pathogenesis of DMED and the mechanism MSCs actions are unclear.Materials and MethodsWe constructed a rat model of DMED with or without intracavernous injection of MSCs, and performed microRNA (miRNA) sequencing of corpora cavernosa tissues.ResultsWe identified three overlapping differentially expressed miRNAs (rno-miR-1298, rno-miR-122-5p, and rno-miR-6321) of the normal control group, DMED group, and DMED+MSCs group. We predicted 285 target genes of three miRNAs through RNAhybrid and miRanda database and constructed a miRNA-target gene network through Cytoscape. Next, we constructed protein-protein interaction networks through STRING database and identified the top 10 hub genes with highest connectivity scores. Five GO terms including cellular response to growth factor stimulus (GO:0071363), ossification (GO:0001503), response to steroid hormone (GO:0048545), angiogenesis (GO:0001525), positive regulation of apoptotic process (GO:0043065), and one Reactome pathway (Innate Immune System) were significantly enriched by 10 hub genes using the Metascape database. We selected the GSE2457 dataset to validate the expression of hub genes and found only the expression of B4galt1 was statistically different (p<0.001). B4galt1 was highly expressed in penile tissues of diabetic rats and would be negatively regulated by rno-miR-1298.ConclusionsThree key miRNAs were identified in DMED rats with stem cell therapy and the miR-1298/B4GalT1 axis might exert function in stem cell therapy for ED.

Mesenchymal Stem Cells Enhance Vascular Endothelial Growth Factor-A, Endothelial Nitric Oxide Synthetase, and HSP70 Expression in Improving Erectile Dysfunction in Streptozotocin-induced Diabetic Rats

This study investigated the therapeutic role of mesenchymal stem cells (MSCs) on erectile function in a diabetes mellitus erectile dysfunction (DMED) rat model by analyzing the expression of endothelial nitric oxide synthetase (eNOS), vascular endothelial growth factor (VEGF), and the 70 kilodalton heat shock proteins (HSP70). MSCs were isolated from umbilical cords (UCs), and their characteristics identified by flow cytometry and osteogenic differentiation analysis. Thirty 8-week-old rats were divided into four groups: sham, control, T1, and T2. After a 16 h fast, 24 rats were randomly selected and intraperitoneally injected with streptozotocin (STZ) to induce DM. At 8 weeks after STZ injection, rats with DMED were classified into four groups, sham, control group [DMED rats received 500 μL phosphate buffer saline (PBS)]; T1 [DMED rats treated with 500 μL PBS containing 1 × 106 UC-MSCs]; T2 [DMED rats treated with 500 μL PBS containing 2 × 106 UC-MSCs]. Eight weeks after MSCs administration, the rats’ erectile function was measured by cavernous nerve stimulation. The blinded histological and gene expression assessment were used to analyze the eNOS, HSP70 content, and VEGF expression on the penile tissues. MSCs administration, rats in T1 and T2 groups showed a significant enhancement of erectile response that showed a trend of increase of VEGF mRNA level expression was 2.2 ± 0.61 in T2 Group supported with the optimum recovery of eNOS, in which the value of eNOS expression was 20.66% ± 2.32%. While optimum decrease of HSP70 content, the value of HSP70 expression was 15.50% ± 0.90%. IHC results showed that the DMED induction in rats caused a significant decrease of eNOS content in corpus cavernosum tissue. MSCs could ameliorate DMED in rats by increasing VEGF and decreasing HSP70 and eNOS, indicating these cells offer a potential application for DMED patients’ treatment.

Knockdown of miR-423-5p simultaneously upgrades the eNOS and VEGFa pathways in ADSCs and improves erectile function in diabetic rats.

This study aimed to explore the possibility of miR‐423‐5p modified adipose‐derived stem cell (ADSCs) therapy on streptozotocin (STZ)‐induced diabetes mellitus erectile dysfunction (DMED) rats. MiR‐423‐5p was knocked down in ADSCs. ADSCs, NC‐miR‐ADSCs and miR‐ADSCs were co‐cultured with human umbilical vein endothelial cells (HUVECs). Normal and high glucose media were supplemented. The supernatant and HUVECs were collected for assessment of eNOS and VEGFa expression, cell proliferation, and apoptosis. HUVECs co‐cultured with ADSCs or miR‐ADSCs exhibited higher eNOS and VEGFa protein expression levels compared to DM groups. MiR‐ADSCs enhanced HUVEC proliferation compared to the ADSCs and NC‐miR‐ADSCs. Lower apoptotic rates were observed when HUVECs were co‐cultured with miR‐ADSCs, compared to ADSCs and NC‐miR‐ADSCs. Fifteen male Sprague‐Dawley (SD) rats aged 12 weeks were induced to develop diabetes mellitus by intraperitoneal injection with STZ, and five healthy SD rats were used as normal controls. Eight weeks after developing diabetes, the rats received ADSCs and miR‐ADSCs via injection into the corpora cavernosa, whereas normal controls and DM controls were injected with saline. Erectile function and histological assessment of penile tissues were performed 8 weeks after injection. The ICP/MAP indicated that erectile function was impaired in the DM rats compared with the normal group. Injection of ADSCs and miR‐ADSCs improved erectile function significantly and was associated with the overexpression of eNOS and VEGFa. MiR‐423‐5p knockdown in ADSCs ameliorated high glucose‐mediated damage to HUVECs and improved erectile function in DM rats by inducing eNOS and VEGFa overexpression, indicating that miR‐423‐5p may be a potential target in the treatment of DMED.

Inhibition of inducible nitric oxide synthase improved erectile dysfunction in rats with type 1 diabetes.

Diabetes mellitus (DM), which is closely related to microvascular dysfunction, is a risk factor for erectile dysfunction (ED). Furthermore, the upregulation of inducible nitric oxide synthase (iNOS) is associated with systemic vascular dysfunction in rats with diabetes. The purpose of this study was to investigate the role of iNOS in diabetes mellitus erectile dysfunction (DMED). First, we developed a type 1 DM rat model using streptozotocin and selected those that developed DMED. Then, we injected these rats with the 1400W, an iNOS inhibitor, for 10weeks and subsequently assessed their ED. Lastly, we performed various molecular studies and histopathological analyses of penile tissues collected from these rats after the experiments. Through the histopathological studies, we also found that the treatment restored the ratios of the smooth muscle to collagen fibres, delayed the development of microvascular injury and alleviated the oxidative stress caused by hyperglycaemia. Based on these results, we confirmed that upregulation of iNOS leads to microvascular dysfunction in patients with ED. Overall, we found that inhibition of iNOS displayed beneficial effects in the treatment of ED, suggesting that its mechanism should be further explored.

Extracorporeal shock wave therapy combined with engineered mesenchymal stem cells expressing stromal cell-derived factor-1 can improve erectile dysfunction in streptozotocin-induced diabetic rats.

BackgroundFor erectile dysfunction (ED) in diabetes mellitus (DM) patients who have poor response to drugs, extracorporeal shock wave therapy (ESWT) and engineered mesenchymal stem cell (MSC) therapy have been studied as alternative treatment options. The objective of this study is to investigate whether ESWT in combination with stromal cell-derived factor-1 expressing engineered mesenchymal stem cell (SDF-1 eMSC) therapy can have synergistic effects on ED in streptozotocin-induced diabetic rats.MethodsFifty 8-week-old male Sprague-Dawley rats were randomly divided into five groups (N=10 per group): (I) Normal group, (II) DM ED, (III) DM ED + ESWT group, (IV) DM ED + SDF-1 eMSC group, and (V) DM ED + ESWT + SDF-1 eMSC group. Each groups were treated with bilateral injections of SDF-1 eMSC or ESWT following the experiment protocol for eight weeks.ResultsThe ratio of ICP/MAP was distinctly higher in the DM ED + ESWT + SDF-1 eMSC group than that in the DM ED group. Concentration of α-smooth muscle actin (α-SMA) was elevated the highest in the DM ED + ESWT + SDF-1 eMSC group. Additionally, ESWT increased the intensity of SDF-1 expression in the corpus cavernosum. ESWT + SDF-1 eMSC treatment also induced neuronal nitric oxide synthase (nNOS) and NO/cGMP expression in the corpus cavernosum. Furthermore, numbers of penile progenitor cells were increased in DM ED rats.ConclusionsCombined treatment of ESWT with SDF-1 eMSC treatment is more effective than by a single therapy. It could be used as a potential and effective synergistic treatment for DM ED.

Identification of key genes in type 2 diabetes-induced erectile dysfunction rats with stem cell therapy through high-throughput sequencing and bioinformatic analysis.

Diabetes mellitus erectile dysfunction (DMED) is a frequent complication of diabetes. Mesenchymal stem cell (MSC) therapy was demonstrated to improve erectile function in DMED. However, the pathogenesis of DMED and the mechanism by which MSCs function are still unclear. We established a rat model of DMED and gave MSC therapy through intracavernous injection. After transcriptome sequencing of rats' penile tissue, we identified a total of 1,097 overlapped differentially expressed genes (DEGs) of the normal control group, DMED group, and MSC-treated group, containing 189 upregulated genes and 908 downregulated genes. The enriched functions of upregulated DEGs included extracellular matrix organisation (GO:0030198), extracellular structure organisation (GO:0043062), and wound healing (GO:0042060), PPAR signalling pathway (rno03320), arachidonic acid metabolism (rno00590) and retinol metabolism (rno00830). The enriched functions of downregulated DEGs included peptidase activity (GO:0052547), hair follicle development (GO:0001942), intermediate filament-based process (GO:0045103), nitrogen metabolism (rno00910), aldosterone-regulated sodium reabsorption (rno04960) and retinol metabolism (rno00830). We constructed a PPI network with 547 nodes and 2,365 edges and identified 15 hub genes with high connectivity degree. In summary, 15 hub genes with potential roles in the development of ED were identified. Further functional research would be required to elucidate the molecular mechanism underlying misregulated genes.

Elevated pigment epithelium-derived factor induces diabetic erectile dysfunction via interruption of the Akt/Hsp90β/eNOS complex.

Diabetes mellitus erectile dysfunction (DMED) is a common complication of diabetes. The level of pigment epithelium-derived factor (PEDF) is significantly upregulated in the serum of individuals with obesity and diabetes. However, whether elevated PEDF levels contribute to DMED remains unknown. This study aimed to investigate the pathogenic role of PEDF and its related mechanism in DMED. We enrolled 65 men, of whom 20 were nondiabetic control participants, 21 participants with diabetes but without erectile dysfunction, and 24 with DMED. The International Index of Erectile Function (IIEF-5) questionnaire was administered to evaluate erectile function. Plasma PEDF in diabetic participants and streptozotocin (STZ)-induced diabetic animals was detected by ELISA. Erectile function was evaluated by measuring the intracavernous pressure (ICP) and the ICP/mean arterial pressure (MAP) ratio in STZ-induced diabetic rats treated with PEDF-neutralising antibody (PEDF-Ab), db/db mice treated with PEDF-Ab, and Pedf knockout mice with STZ-induced diabetes. The overexpression of PEDF was implemented by intraperitoneal injection of recombinant PEDF and intracavernous injection of PEDF-expressing adenovirus. A mechanistic study was performed by immunofluorescence staining, bimolecular fluorescence complementation (BiFC), immunoprecipitation and western blotting. We found that the plasma level of PEDF was significantly higher in participants with DMED compared with diabetic counterparts without erectile dysfunction and nondiabetic controls. Interestingly, PEDF levels were negatively correlated with plasma nitrite/nitrate levels and erectile function in DMED patients and STZ-induced diabetic rats. Furthermore, overexpression of PEDF significantly suppressed ICP and endothelial nitric oxide synthase (eNOS) phosphorylation in control rats. In contrast, the PEDF-Ab and Pedf knockout ameliorated ICP and eNOS phosphorylation in diabetic rats and mice. Mechanistically, PEDF promoted the membrane translocation of Hsp90β and directly bound to the amino acid residues 341-724 of Hsp90β on the endothelial cell surface, subsequently blocking intracellular Hsp90β/Akt/eNOS complex formation and downregulating eNOS phosphorylation. These results indicate that elevated PEDF levels contribute to impaired erectile function by suppressing Hsp90β-mediated eNOS phosphorylation and that PEDF may represent a novel therapeutic target for diabetic erectile dysfunction. Graphical abstract.

Effect of icariside II and metformin on penile erectile function, glucose metabolism, reaction oxygen species, superoxide dismutase, and mitochondrial autophagy in type 2 diabetic rats with erectile dysfunction.

BackgroundIcariside II (ICAII) is a flavonoid isolated from herb Epimedium that has been shown to improve erectile function in rats. However, ICAII’s underlying mechanism remains unclear.MethodsType 2 diabetes mellitus erectile dysfunction (T2DMED) rats were induced by single intraperitoneal injection of 25 mg/kg streptozotocin (STZ) and fed a high-fat, high-sugar, and high-calorie diet for 8 weeks. In the control and T2DMED groups, rats were administered with normal saline; in the metformin (MET) group, rats were administered with MET at 0.2 g/kg/day; and in the ICAII + MET group, rats were administered with ICA II at 10 mg/kg/day and MET for 0.2 g/kg/day. The deposition of advanced glycation end products (AGEs), expression of receptor for AGEs (RAGEs), reactive oxygen species (ROS), and superoxide dismutase (SOD) activity, and corpus cavernosum smooth muscle cells (CCSMCs) mitochondrial autophagy were measured. We also evaluated the expression of LC3-II/I, Beclin-1, P70S6K, PI3K, AKT, mTOR, phospho-AKT, phospho-mTOR, and phospho-P70S6K.ResultsICAII and MET can improve erectile function, and decrease the levels of fasting plasma glucose (FPG), hemoglobin A1c (HbA1c), and AGEs in rats with T2DMED. Furthermore, ICAII and MET can decrease excessive CCSMC mitochondrial autophagy and the level of RAGE and oxidant stress in rats with T2DMED. ICAII and MET may enhance signaling via the PI3K-AKT-mTOR pathway in order to reduce the excessive mitochondrial autophagy of CCSMCs.ConclusionsICAII may effectively improve penile erectile function via decreasing excessive CCSMCs mitochondrial autophagy, RAGE, and oxidant stress. Furthermore, ICAII may enhance signaling via the PI3K-AKT-mTOR pathway in order to reduce excessive CCSMC mitochondrial autophagy.

Effect of Icariside II and Metformin on Penile Erectile Function, Histological Structure, Mitochondrial Autophagy, Glucose-Lipid Metabolism, Angiotensin II and Sex Hormone in Type 2 Diabetic Rats With Erectile Dysfunction.

IntroductionType 2 diabetes mellitus erectile dysfunction (T2DMED) is one of the common complications of type 2 diabetes mellitus (T2DM). Icariside II (ICA II), a flavonoid derived from Epimedium, has been shown to improve erectile function in T2DMED rats.AimTo investigate the effect of ICA II and metformin (MET) on penile erectile function, mitochondrial autophagy, glucose-lipid metabolism in rats with T2DMED.MethodsIn the control and T2DMED groups, rats were administered normal saline. In the MET group, rats were administered MET for 0.2 g/kg/day. In the ICA II+MET group, rats were administered ICA II for 10 mg/kg/day and MET for 0.2 g/kg/day.ResultsThe number of mating rats, number of erectile rats, erection rate, erection frequency, intracorneal pressure, and intracorneal pressure/mean arterial pressure in the ICA II+MET group and control group were significantly higher than corresponding values in than T2DMED group. The absolute values of fasting plasma glucose, glycated haemoglobin in the ICA II+MET group, MET group, and control group were significantly lower than in the T2DMED group. The advanced glycation end product (AGE) values in the ICA II+MET group and the MET group were lower than in the T2DMED group. The receptors for the AGE values and angiotensin II values in the ICA II+MET group were lower than in the T2DMED and MET groups. The high-density lipoprotein values, testosterone values, nitric oxide synthase activity, and cyclic guanosine monophosphate content in the ICA II+MET and control groups were higher than in the T2DMED group. The low-density lipoprotein values, triglyceride values, estradiol values, and total cholesterol values in the ICA II+MET and control groups were lower than in the T2DMED group.ConclusionICA II could increase erectile function and smooth muscle cell/collagen fibril proportions, decreased mitochondrial autophagy, and AGE concentrations and improve lipid metabolism, nitric oxide synthase activity, cyclic guanosine monophosphate content, testosterone, estradiol, and Ang II in rat with T2DMED.Zhang J, Li S, Zhang S, et al. Effect of Icariside II and Metformin on Penile Erectile Function, Histological Structure, Mitochondrial Autophagy, Glucose-Lipid Metabolism, Angiotensin II and Sex Hormone in Type 2 Diabetic Rats With Erectile Dysfunction. J Sex Med 2020;8:168–177.

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.

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).

Epigenetic silencing of microRNA-874-3p implicates in erectile dysfunction in diabetic rats by activating the Nupr1/Chop-mediated pathway.

Diabetes is a global medical problem that causes many deaths every year. Complications caused by diabetes are serious and affect patients' quality of life. Diabetes mellitus erectile dysfunction (DMED) affects more than half of male diabetes patients. In this study, we determined the role of microRNA-874-3p (miR-874-3p) and nuclear protein-1 (Nupr1) in streptozocin-induced DMED rats. Control rats received equal amount of vehicle. These rats were also injected with lentiviral vector or agomir to silence or overexpress miR-874-3p or Nupr1. Apomorphine (100μg/kg, s.c.) was used to induce erection and time of erection was recorded. Intracavernosal and mean arterial pressure ratio (ICP/MAP) were also recorded. O2- level and concentration of thiobarbituric acid reactive substances (TBARs) were detected using lucigenin-derived chemiluminescence method and Colorimetry. Rat cavernosum tissues were collected for subsequent experiments. Cavernosum smooth muscle cells (CSMCs) were also used for in vitro experiments. Nupr1 was found highly expressed (by RT-qPCR and Western blot analysis) in cavernosum tissues from DMED rats. Nupr1 silencing improved the ICP/MAP ratio and erection time. Nupr1 silencing also reduced CSMC apoptosis (by TUNEL assay) as well as decreased O2- level and TBAR concentration. Nupr1 was targeted and inhibited by miR-874-3p (by luciferase activity and RNA immunoprecipitation assays), which was downregulated in DMED. miR-874-3p downregulation was due to increased methylation at the promoter region (methylation-specific PCR). miR-874-3p overexpression improved erection time and reduced apoptosis. In summary, miR-874-3p was downregulated which led to increased apoptosis and erectile dysfunction in DMED rats, through inhibition of Nupr1-mediated pathway. This study may also provide a new therapeutic direction for the treatment of DMED.