Effect of testosterone and frequent low‐dose sildenafil/tadalafil on cavernous tissue oxidative stress of aged diabetic rats

This study aimed to assess the effect of testosterone (T) administration and chronic low‐dose sildenafil/tadalafil on cavernous tissue oxidative stress (OS) of aged diabetic rats. In all, 140 Sprague‐Dawley aged rats were subdivided into the following: controls; streptozotocin (STZ)‐induced diabetic rats; diabetic rats injected with T every 4 weeks; diabetic rats on sildenafil orally daily; diabetic rats on T and daily sildenafil; diabetic rats on tadalafil orally every other day; diabetic rats on T and tadalafil; diabetic rats on alternate sildenafil/tadalafil; and diabetic rats on alternate sildenafil/tadalafil with T. After 12 weeks, the rats were euthanised where in dissected cavernous tissues malondialdehyde (MAD), glutathione peroxidase (GPx) and cGMP (cyclic guanosine monophosphate) were estimated. Compared with controls, aged diabetic rats demonstrated significant increase in cavernous tissue MDA and significant decrease in GPx and cGMP where diabetic rats injected with T had marked improvement of these parameters. Diabetic rats on sildenafil, tadalafil or alternate sildenafil/tadalafil demonstrated significant increased cavernous tissue GPx, cGMP and decreased cavernous MDA that was further improved when supplemented with T. It is concluded that frequent low‐dose use of sildenafil and/or tadalafil supplemented with T has a marked impact on ameliorating cavernous OS in aged diabetic rats.     

Favourable effect of β‐glucan treatment against cisplatin‐induced reproductive system damage in male rats

The aim of this study was to investigate the potential beneficial effects of β‐glucan treatment against oxidative, histological and spermatological damage caused by cisplatin on the male reproductive system. Twenty‐eight Sprague Dawley male rats were used in the study. The rats were randomly divided into four equal‐sized groups: a control group, cisplatin group (7 mg/kg in a single‐dose cisplatin administered intraperitoneally), β‐glucan group (β‐glucan given at a dose of 50 mg kg^?1 d^?1 for 14 day) and a cisplatin plus β‐glucan group (cisplatin and β‐glucan administered together at the same dose). Cisplatin administration induced an increase in the level of thiobarbituric acid‐reactive substances, a lipid peroxidation indicator. It induced a decrease in enzymatic (superoxide dismutase, catalase and glutathione peroxidase) activities and nonenzymatic (reduced glutathione) antioxidant levels. In addition, cisplatin caused both histological and spermatological damage, as shown by a decrease in sperm motility and epididymal sperm concentrations and an increase in abnormal sperm rates. The β‐glucan treatment improved cisplatin‐induced oxidative, histological and spermatological damage. This study revealed that β‐glucan treatment provided prevention against male reproductive system damage caused by cisplatin. These preventative effects were likely due to its antioxidant properties.     

Safety and efficacy of sildenafil citrate in treating erectile dysfunction in patients with combat‐related post‐traumatic stress disorder: a double‐blind,randomized and placebo‐controlled study

OBJECTIVE

To evaluate the safety and efficacy of sildenafil citrate for treating erectile dysfunction (ED) in patients with combat‐related post‐traumatic stress disorder (PTSD).

PATIENTS AND METHODS

In all, 266 combat‐exposed war veterans with ED (aged 37–59 years) were recruited. They met the Diagnostic and Statistical Manual of Mental Disorders‐IV criteria for PTSD according to the Structured Clinical Interview for Patients, Investigator Version. The patients were also evaluated with the Clinician‐Administered PTSD Scale, both to establish the diagnosis of PTSD and to measure symptom severity. Only patients with psychogenic ED were included in the study. Patients with comorbid conditions (diabetes mellitus, hypercholesterolaemia, hypertension, Peyronie’s disease) and smokers of more than five cigarettes daily were excluded. The patients were randomly divided into a group of 133 who received 100 mg of on‐demand sildenafil 0.75–2 h before sexual stimulation, and 133 who received placebo. Patients were asked to use ≥16 doses or attempts at home. The efficacy of the treatments was assessed every four attempts during treatment, and at the end of the study, using responses to the 15‐question International Index of Erectile Function (IIEF), Sexual Encounter Profile diary questions 2 and 3, Erectile Dysfunction Inventory of Treatment Satisfaction questionnaire, patients’ event logs of sexual activity, and a Global Assessment Question about erections.

RESULTS

Sildenafil did not produce significantly and substantially greater improvement than placebo in each of the primary and secondary outcome measures (P = 0.08). A normal EF domain score (≥26) at endpoint was reported by 13 (9.8%), and 11 (8.3%) of patients on the sildenafil and placebo regimens, respectively (P = 0.09). Patients treated with sildenafil had no statistically significantly greater improvement in the five sexual function domains of the IIEF questionnaire than those treated with placebo (P = 0.08). The incidences of treatment‐emergent adverse events were significantly greater in the sildenafil arm than in the placebo group (P = 0.01).

CONCLUSIONS

Sildenafil is no better than placebo in treating PTSD‐emergent ED. Further randomized clinical trials are warranted in combat veterans and other populations with PTSD to better elucidate the role of phosphodiesterase type 5 inhibitors in treating PTSD‐emergent ED.     

Aminoguanidine prevents testicular damage–induced‐2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD) in male rats

In this study, it was aimed to determinate protective effects of aminoguanidine (AG) against reproductive toxicity caused by 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin (TCDD), an environmental contaminant. Thirty‐two rats were equally divided into four groups; the first group was kept as control and given corn oil as carrier. In second and third groups, TCDD and AG were orally administered at the dose of 2 μg kg^?1 per week and 100 mg kg^?1 per day for 45 days, respectively. In fourth group, TCDD and AG were given together at the same doses. Although TCDD significantly increased the formation of TBARS, it caused a significant decline in the levels of GSH, CAT, GPx and SOD in rats. On the other hand, AG, given together TCDD, reversed TCDD effects on TBARS SOD, GSH, GPx and CAT. In addition, sperm characteristics negatively affected and histopathological deformation occurred with TCDD exposure. However, AG treatment partly prevented these toxic effects of TCDD on spermatological parameters and histopathological changes. In conclusion, TCDD exposure induces testicular damage (oxidative stress, histopathological damage and sperm parameters), and AG treatment reversed TCDD‐induced testicular damage in rats. Thus, AG may be useful for the prevention and treatment of TCDD‐induced male infertility problems.     

Protective role of Diospyros lotus on cisplatin‐induced changes in sperm characteristics,testicular damage and oxidative stress in rats

The aim of this study was to investigate the protective effect of Diospyros lotus (DL) on cisplatin (CP)‐induced testicular damage in male rats. Twenty‐eight male rats were randomly divided into four groups: group 1 – control, given isotonic saline solution; group 2 – CP 7 mg kg⁻¹ given intraperitoneally as single dose; group 3 – DL 1000 mg kg⁻¹ per day given orally for 10 days; group 4 – CP and DL given together at the same doses. CP caused a significant increase in thiobarbituric acid‐reactive substances (TBARS) level and a significant decrease in superoxide dismutase (SOD), glutathione peroxidase (GPx), catalase (CAT) and glutathione (GSH) levels in rats testis tissues compared to the control group. CP caused a significant increase in lipid peroxidation in testis tissues compared to the control group, whereas DL led to a significant increase in SOD and GSH levels. However, there were no statistically significant changes in GPx and CAT levels. In addition, serum testosterone levels, sperm concentration and sperm motility were significantly decreased, but abnormal sperm rate and histological changes were increased with CP. However, these effects of CP on sperm parameters, histological changes and the tissue weights were eliminated by DL treatment. In conclusion, our study showed that the reproductive toxicity caused by CP may be prevented by DL treatment.     

Evaluation of 5α‐reductase inhibitory activity of certain herbs useful as antiandrogens

This study demonstrates 5α‐reductase inhibitory activity of certain herbs useful in the management of androgenic disorders. Ganoderma lucidum (Curtis) P. Karst (GL), Urtica dioica Linn. (UD), Caesalpinia bonducella Fleming. (CB), Tribulus terrestris Linn. (TT), Pedalium murex Linn. (PM), Sphaeranthus indicus Linn. (SI), Cuscuta reflexa Roxb. (CR), Citrullus colocynthis Schrad. (CC), Benincasa hispida Cogn. (BH), Phyllanthus niruri Linn. (PN) and Echinops echinatus Linn. (EE) were included in the study. Petroleum ether, ethanol and aqueous extracts of these herbs were tested for their 5α‐reductase inhibitory activity against the standard 5α‐reductase inhibitor, finasteride. A biochemical method to determine the activity of 5α‐reductase was used to evaluate the inhibition of different extracts to the enzyme. The optical density (OD) value of each sample was measured continuously with ultraviolet spectrophotometer for the reason that the substrate NADPH has a specific absorbance at 340 nm. As the enzyme 5α‐reductase uses NADPH as a substrate, so in the presence of 5α‐reductase inhibitor, the NADPH concentration will increase with the function of time. This method thus implicates the activity of 5α‐reductase. The method proved to be extremely useful to screen the herbs for their 5α‐reductase inhibitory potential. GL, UD, BH, SI and CR came out to be promising candidates for further exploring their antiandrogenic properties.     

Combined effects of TNF‐α, IL‐1β, and HIF‐1α on MMP‐2 production in ACL fibroblasts under mechanical stretch: An in vitro study

The dynamics between inflammatory factors, mechanical stress, and healing factors, in an intra‐articular joint, are very complex after injury. Injury to intra‐articular tissue [anterior cruciate ligament (ACL), synovium] results in hypoxia, accumulation of various pro‐inflammatory factors, cytokines, and metalloproteases. Although the presence of increased amounts of matrix‐metalloproteinases (MMP) in the joint fluid after knee injury is considered the key factor for ACL poor healing ability; however, the exact role of collective participants of the joint fluid on MMP‐2 activity and production has not been fully studied yet. To investigate the combined effects of mechanical injury, inflammation and hypoxia induced factor‐1α (HIF‐1α) on induction of MMP‐2; we mimicked the microenvironment of joint cavity after ACL injury. The results show that TNF‐α and IL‐1β elevate the activity of MMP‐2 in a dose‐ and time‐dependent manner. In addition, mechanical stretch further enhances the MMP‐2 protein levels with TNF‐α, IL‐1β, and their mixture. CoCl₂‐induced HIF‐1α (100 and 500 µM) also increases the levels and activity of MMP‐2. Mechanical stretch has a strong additional effect on MMP‐2 production with HIF‐1α. Our results conclude that mechanical injury, HIF‐1α and inflammatory factors collectively induce increased MMP‐2 production in ACL fibroblasts, which was inhibited by NF‐κB pathway inhibitor (Bay‐11‐7082). © 2011 Orthopaedic Research Society Published by Wiley Periodicals, Inc. J Orthop Res 29: 1008–1014, 2011     

Efficacy of tamoxifen and l‐carnitine on sperm ultrastructure and seminal oxidative stress in patients with idiopathic oligoasthenoteratozoospermia

Idiopathic oligoathenoteratozoospermia (iOAT) is a common finding in the evaluation of male infertility. Oxidative stress (OS) may underlie its pathology. Tamoxifen and l ‐carnitine are used to treat idiopathic male infertility. The aim of this work was to detect the efficacy of tamoxifen and l ‐carnitine on sperm parameters, sperm ultrastructure and seminal OS in iOAT patients. Sixty patients were recruited for this study and divided into three groups; the 1st was treated with tamoxifen, 2nd with l ‐carnitine and 3rd with both drugs. Semen analysis, malondialdehye (MDA) level and transmission electron microscopy were performed before and after three months treatment. The first group showed significant improvement in MDA levels, sperm concentration, sperm morphology, ultrastructural head, acrosomal and mitochondrial anomalies (P < 0.01). Other parameters were not significantly improved. In the 2nd group, significant improvements in MDA, sperm motility, sperm morphology, ultrastructural mitochondrial and tail anomalies were detected (P < 0.01). No significant improvement in the other parameters. Third group showed improvement in MDA, all semen parameters and all ultrastructural anomalies (P < 0.01). In conclusion, tamoxifen and l ‐carnitine are effective in improving seminal OS, semen parameters and sperm ultrastructure. Combination of both drugs is superior to monotherapy.     

Proteomic identification of 14‐3‐3ϵ as a linker protein between pERK1/2 inhibition and BIM upregulation in human osteosarcoma cells

Despite advancements in multimodality chemotherapy, conventional cytotoxic treatments still remain ineffective for a subset of patients with aggressive metastatic or multifocal osteosarcoma. It has been shown that pERK1/2 inhibition enhances chemosensitivity to doxorubicin and promotes osteosarcoma cell death in vivo and in vitro. One of the pro‐apoptotic mechanisms is upregulation of Bim by pERK1/2 inhibitors. To this end, we examined proteomic changes of 143B human osteosarcoma cells with and without treatment of PD98059, pERK1/2 inhibitor. Specifically, we identified 14‐3‐3? protein as a potential mediator of Bim expression in response to inhibition of pERK1/2. We hypothesized that 14‐3‐3? mediates upregulation of Bim expression after pERK1/2 inhibition. We examined the expression of Bim after silencing 14‐3‐3? using siRNA. The 14‐3‐3? gene silencing resulted in downregulation of Bim expression after PD98059 treatment. These data indicate that 14‐3‐3? is required for Bim expression and that it has an anti‐cancer effect under pERK1/2 inhibition in 143B cells. By playing an essential role upstream of Bim, 14‐3‐3? may potentially be a coadjuvant factor synergizing the effect of pERK1/2 inhibitors in addition to conventional cytotoxic agents for more effective osteosarcoma treatments. © 2014 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 32:848–854, 2014.

     

Inhibitory activity of α‐tocoferol on apoptosis in the rat bladder wall subjected to androgen deprivation

Alpha‐tocopherol (2,5,7,8‐tetramethyl‐2‐(4,8,12‐trimethyltridecyl)‐chromon‐8‐ol) is used in many previous urological studies. Thus to add to this knowledge in this study we studied the potential inhibitory activity on oxidative stress and process apoptosis on bladder wall in male rats subjected to androgen deprivation. A causal relationship between lower testosterone levels and apoptosis, as a component of castration‐induced muscle atrophy, has been shown. Male Wistar rats (250–300 g) were used in this experiment, divided into four groups: control (sham operation; n = 10); castration (n = 10); castration, with alpha‐tocopherol supplementation over preceding four weeks (n = 10); and castration, with alpha‐tocopherol supplementation over preceding four weeks and subsequent eight weeks (n = 10). Activated caspase‐3 was detected using a previously described technique, with analysis using stereological methodology. Nonparametric methods were used to test statistical significance, taking a significance level of P < 0.005. This study was approved by the ethics committee of the university where the project was developed. The serum testosterone concentrations before castration were less than 20 pg/ml. Analysis of 8‐isoprostane showed statistical significance (P < 0.0003). The volumetric density of caspase‐3 showed significant differences between the groups. There was no statistical significance regarding caspase‐3 between sham and alpha‐tocopherol plus castration or between the groups that received alpha‐tocopherol supplementation. The observations showed that there was greater apoptosis in the group with castration alone than in the groups with alpha‐tocopherol supplementation. This finding, together with the induced androgen deprivation and higher 8‐isoprostane levels, corroborates the hypothesis that alpha‐tocopherol supplementation has an important protective effect under conditions of oxidative stress, thereby avoiding the apoptotic process, especially regarding aging. Neurourol. Urodyn. 30:194–198, 2011. © 2010 Wiley‐Liss, Inc.     

Cytoprotective effects of fruit pulp of Eugenia jambolana on H 2 O 2‐induced oxidative stress and apoptosis in rat Leydig cells in vitro

This study was undertaken to investigate the cytoprotective effect of the fruit pulp of Eugenia jambolana (50–250 μg ml^?1) against the damage induced by H ₂ O ₂ (100 μm ) exposure to Leydig cells in vitro. Cell survival with extract was found comparable to similar effects by N‐acetyl‐l ‐cysteine. H ₂ O ₂‐induced rise in thiobarbituric acid reactive substance formation and decline in the activity and expression of antioxidant enzymes like superoxide dismutase, catalase and glutathione‐s‐transferase were effectively checked. Cellular glutathione and total antioxidant capacity demonstrated significant improvement. The increase in expression of inducible nitric oxide (NO) synthase leading to NO production was successfully countered. Co‐treatment of the extract helped in the down‐regulation of caspase‐3 and poly‐ADP‐ribose polymerase resulting in a significant reduction in Leydig cell apoptosis induced by H ₂ O ₂. Upstream marker proteins of extrinsic (caspase‐8, Fas, FasL) and intrinsic (caspase‐9) pathway of metazoan apoptosis were identically down‐regulated. The Bcl‐2 family of proteins, though, remained unaffected. The extract also positively modulated the other marker proteins like c‐Jun NH ₂‐terminal kinase, p38, Akt, nuclear factor‐κB, c‐Fos, cellular FLICE‐inhibitory protein, cyclooxygenase‐2 and p53. Taken together, the above‐mentioned findings establish the anti‐oxidative and anti‐apoptotic potency of the extract that ameliorates the H ₂ O ₂‐induced adverse effects on rat Leydig cells in vitro.     

Increased gene expression of TGF‐β in peripheral blood mononuclear cells from renal transplant patients with polyomavirus BK viremia

We aimed to investigate the roles of cytokines during polyomavirus BK (BKV) reactivation in renal transplant patients. Forty‐eight renal allograft recipients were enrolled, and their sera BKV viral load and mRNA expression levels of cytokines in peripheral blood mononuclear cells were measured by real‐time polymerase chain reaction. Patient's age and gene expression levels of interleukin (IL)‐2 (10.04 ± 2.63 vs. 8.70 ± 2.40, p = 0.049) and transforming growth factor (TGF)‐β (12.58 ± 2.59 vs. 10.89 ± 1.91, p = 0.015) were significantly higher in BKV viremia (+) renal transplant patients. Multivariate logistic regression analysis revealed that age and mRNA expression levels of TGF‐β, but not IL‐2, significantly correlated with the presence of BKV viremia. Sera BKV viral loads showed a positive correlation with patient age and the levels of TGF‐β and IL‐6 mRNA. After adjusting for age and sex in the regression model, both age and TGF‐β mRNA levels maintained a significant positive association with sera BKV viral loads. Serum TGF‐β concentration tended to be higher in BKV viremia (+) patients (p = 0.079). In conclusion, expression levels of TGF‐β were found to correlate with both BKV viremia positivity and sera BKV viral loads in renal transplant patients.     

Indirubin‐3′‐Oxime Reverses Bone Loss in Ovariectomized and Hindlimb‐Unloaded Mice Via Activation of the Wnt/β‐Catenin Signaling

Osteoporosis is a major global health issue in elderly people. Because Wnt/β‐catenin signaling plays a key role in bone homeostasis, we screened activators of this pathway through cell‐based screening, and investigated indirubin‐3′‐oxime (I3O), one of the positive compounds known to inhibit GSK3β, as a potential anti‐osteoporotic agent. Here, we show that I3O activated Wnt/β‐catenin signaling via inhibition of the interaction of GSK3β with β‐catenin, and induced osteoblast differentiation in vitro and increased calvarial bone thickness ex vivo. Intraperitoneal injection of I3O increased bone mass and improved microarchitecture in normal mice and reversed bone loss in an ovariectomized mouse model of age‐related osteoporosis. I3O also increased thickness and area of cortical bone, indicating improved bone strength. Enhanced bone mass and strength correlated with activated Wnt/β‐catenin signaling, as shown by histological analyses of both trabecular and cortical bones. I3O also restored mass and density of bone in hindlimb‐unloaded mice compared with control, suspended mice, demonstrating bone‐restoration effects of I3O in non‐aged–related osteoporosis as well. Overall, I3O, a pharmacologically active small molecule, could be a potential therapeutic agent for the treatment and prevention of osteoporosis. © 2014 American Society for Bone and Mineral Research.