Castration Group Research Articles

Low androgen status inhibits erectile function by inducing eNOS uncoupling in rat corpus cavernosum.

The number of erectile dysfunction (ED) patients is increasing annually. How to improve the effectiveness of ED treatment is an important issue for the field of andrology. To investigate whether low androgen status impairs the erectile function of rats by regulated endothelial nitric oxide synthase (eNOS) uncoupling. Thirty-six 8-week-old male Sprague Dawley (SD) rats were randomly divided into six groups as follows: 4-week sham-operated group (4w-sham), 4-week castration group (4w-cast), 4-week castration+testosterone (T) group (4w-cast+T), 8-week sham-operated group (8w-sham), 8-week castration group (8w-cast), and 8-week castration+T group (8w-cast+T). Three mg/kg of T was subcutaneously injected every other day in castration+T groups. The ratio of the maximum intracavernous pressure/the mean arterial pressure (ICPmax/MAP), the level of serum T, dihydrobiopterin(BH2 ), tetrahydrobiopterin (BH4 ), nitric oxygen(NO), 3-nitrotyrosine(3NT), dihydrofolate reductase (DHFR), guanosine triphosphate cyclohydrolase 1 (GTPCH1), nicotinamide adenine dinucleotide phosphate oxidase 2 (NOX2), and eNOS monomers/dimers in the corpus cavernosum were detected. The ratio of ICPmax/MAP and BH4 /BH2 , the level of serum T, NO, and GTPCH1 decreased significantly in castration groups compared with sham-operated groups and castration+T groups (P<.05) and decreased significantly in 8w-cast group compared with 4w-cast group (P<.05). The expression of 3NT and NOX2 and the ratio of eNOS monomers/dimers increased significantly in castration groups compared with sham-operated groups and castration+T groups (P<.01) and increased significantly in 8w-cast group compared with 4w-cast group (P<.01). The expression of DHFR in 4w-cast group was significantly higher than that in 4w-sham group and 4w-cast+T group (P<.01) and in 8w-cast group was significantly lower than that in 8w-sham group and 8w-cast+T group (P<.01). Low androgen status induces eNOS uncoupling by reducing BH4 /BH2 and increasing 3NT. Due to the decreased NO production, the erectile function of the rats was impaired.

Integrated lipidomics and targeted metabolomics analyses reveal changes in flavor precursors in psoas major muscle of castrated lambs

Castration may decrease off-odors and improve meat flavor. Meat flavor is generated through complex chemical reactions that involve hydrophilic and hydrophobic flavor precursors. In this study, we investigated the flavor precursors in psoas major muscles of castrated and intact sheep using lipidomics and targeted metabolomics. Castration decreased testosterone levels and increased intramuscular fat content. Six hundred fourteen lipid molecules confirmed showed a separation between castrated and intact sheep based on principal component analysis. Fourteen lipid species and 224 lipid molecules increased in castrated sheep. Targeted metabolomics analysis showed that 18 hydrophilic metabolites were affected by castration; however, only hypoxanthine significantly increased in the castration group. Among 45 volatiles identified, 1-octen-3-ol and hexanal were significantly higher in castrated sheep. These results revealed that lipids, hydrophilic metabolites, and volatile compounds in lamb were affected by castration, which might be beneficial in lamb quality.

Effect of androgen on bone metabolism in hyperuricemic rats.

IntroductionThis study aimed to investigate the effect of androgen on bone metabolism in hyperuricemic rats.Material and methodsForty male Wistar rats were randomly divided into four groups: sham operation group, simple hyperuricemic group, hyperuricemic castration group, and simple castration group. A rat model of chronic hyperuricemia was established using potassium oxonate and ethambutol. Blood was sampled from the vena angularis at week 0, 4, 6, 8 and 12 after surgery to detect for uric acid, calcium, phosphorus and alkaline phosphatase, and investigate the effect of androgen on bone metabolism in hyperuricemic rats.ResultsFrom the 4th week, compared with the sham operation group, the differences in uric acid levels between the simple hyperuricemic group and hyperuricemia castration group were statistically significant (p < 0.05), suggesting the successful establishment of the model of hyperuricemia. At the 6th week, uric acid levels decreased in the two hyperuricemic groups, and the difference from the sham operation group decreased (p = 0.05), showing that the modeling method had deteriorated. At the 8th week, the differences in uric acid levels between the two castration groups and sham operation group were statistically significant (p < 0.05). At the 12th week, the differences in serum levels of phosphorus between the simple hyperuricemic group and hyperuricemic castration group were statistically significant (p < 0.05).ConclusionsAndrogen can induce bone metabolism changes in rats with hyperuricemia.

Castration-induced stromal remodeling disrupts the reconstituted prostate epithelial structure

AbstractThe normal prostate epithelial structure is maintained by homeostatic interactions with smooth muscle cells. However, structural alterations of the stroma are commonly observed in prostatic proliferative diseases, leading to the abnormalities of prostate epithelial structure. A decrease in the androgen level experimentally induces stromal remodeling, i.e., replacement of smooth muscle cells with fibroblasts or myofibroblasts. In this study, we investigated the effects of castration-induced stromal remodeling and subsequent aberrant activation of epithelial–stromal interactions on the reconstituted human prostate-like epithelial structure. We performed in vivo experiments using the human prostate epithelial cell line BPH-1 and fetal rat urogenital sinus mesenchyme to generate heterotypic tissue recombinants that form human prostate-like epithelial structure (i.e., solid- and canalized-epithelial cords). Host mice were castrated at 12 weeks post transplantation (castration) and implanted with a dihydrotestosterone pellet at 14 days post castration (androgen replacement treatment; ART). In the castration group, the percentages of fibrotic area and disrupted prostate epithelial structure without the basement membrane (BM) increased proportionally in a time-dependent manner, but were suppressed by ART. In the castration group, tenascin-C (TNC)-positive fibroblasts were abundant in the stroma surrounding disrupted prostate epithelial structure without the BM. TGF-β1 secretion from BPH-1 cells was increased by co-culturing with human primary cultured prostate fibroblasts. TNC mRNA expression was increased in fibroblasts co-culturing with BPH-1 cells and was suppressed by treatment with a TGF-β RI kinase inhibitor. Moreover, in the castration group, the percentage of p-Smad2-positive cells was significantly higher in the stroma surrounding disrupted prostate epithelial structure without the BM. Our results demonstrate that castration-induced stromal remodeling disrupted the reconstituted human prostate-like epithelial structure and induced the appearance of TNC-positive fibroblasts accompanied by activation of TGF-β signaling. The alteration of prostate stromal structure may be responsible for loss of the BM and epithelial cell polarity.

Low androgen status inhibits erectile function by up-regulating the expression of P2X receptors in rat corpus cavernosum.

The aim of our study was to investigate whether low androgen level inhibits the erectile function of rats by regulating the expression of P2X receptors. Thirty-six 8-week-old male SD rats were randomly divided into six groups: sham-operated groups (4w-sham, 8w-sham), castration groups (4w-cast, 8w-cast) and androgen replacement after castration groups (4w-cast+T, 8w-cast+T). The maximum intracavernous pressure/mean arterial pressure (ICPmax/MAP), the levels of serum testosterone (T) and nitric oxide (NO), and the expression of P2X1, P2X2, P2X3, eNOS, p-eNOS, ROCK1 and ROCK2 in the cavernous tissue of rats were determined. The serum T, ICPmax/MAP and NO levels in penile corpus cavernosum in the castration groups were significantly lower than those in other groups (p<.01). The protein expression of P2X1, P2X2, P2X3, ROCK1 and ROCK2 in the castration groups was significantly higher than those in other groups (p<.01). P-eNOS/eNOS of the castration groups were significantly lower than those of other groups (p<.01). The serum T level was negatively correlated with the expression of P2X1, P2X2 and P2X3 in the corpus cavernosum. Low androgen level inhibits erectile function by up-regulating the expression of P2X1, P2X2, P2X3 and RhoA/Rho-kinase resulting in reducing the ratio of p-eNOS/eNOS and the level of NO in corpus cavernosum of rats.

Does the Addition of Abiraterone to Castration Affect the Reduction in Bone Mineral Density?

The in vivo effect of abiraterone on bone mineral density (BMD) in addition to androgen deprivation therapy was examined using a murine model. The mice were separated into the following groups: control, abiraterone, castration, and castration+abiraterone. The percentage change in the ratio of bone to tissue volume (BV/TV), number of osteoblasts and osteoclasts, and the serum level of bone markers were compared on day 21. The BV/TV ratio of the abiraterone, castration, and castration+abiraterone groups was lower than that of the control group. However, the change in the BV/TV ratio in the castration+abiraterone group was not significantly different from that in the castration group. There was no significant difference in the serum TRAP5b level and the number of osteoclasts and osteoblasts between the castration+abiraterone and the castration groups. The addition of abiraterone to castration did not affect BMD in the murine model.

Effects of Enzymatic Hydrolysate of Oyster on Erectile Function in Male Hemicastrated Rats

Oysters have been thought to significantly improve the male sexual function for thousands of years. To explore the effects of an enzymatic hydrolysate of oyster preparation on erectile dysfunction (ED), hemicastrated male rats were divided into control, positive control and oyster hydrolysate high dose group (0.617 g/kg body weight per day), medium dose group (0.308 g/kg) and low dose group (0.103 g/kg), respectively. After castration, the growth, development and the organ indexes of accessory sex glands and immune organs of the rats were significantly decreased than those of normal rats. Oyster hydrolysate significantly shortens the latency of penile erection induced by electrical stimulation in a dose-dependent manner, indicating that oyster hydrolysate also has significant effects on alleviating ED. Further analysis revealed that the serum levels of testosterone and luteinizing hormone and NO were significantly higher than those in the castrated model group, and were similar to those in the normal control group, indicating that oyster hydrolysate promotes the synthesis of testosterone by stimulating the synthesis of luteinizing hormone, which then increased the serum NO content through the NO-cGMP signaling pathway.

Effects of testosterone suppression, hindlimb immobilization, and recovery on [3H]ouabain binding site content and Na+, K+-ATPase isoforms in rat soleus muscle.

We investigated the effects of testosterone suppression, hindlimb immobilization, and recovery on skeletal muscle Na+,K+-ATPase (NKA), measured via [3H]ouabain binding site content (OB) and NKA isoform abundances (α1-3, β1-2). Male rats underwent castration or sham surgery plus 7 days of rest, 10 days of unilateral immobilization (cast), and 14 days of recovery, with soleus muscles obtained at each time from cast and noncast legs. Testosterone reduction did not modify OB or NKA isoforms in nonimmobilized control muscles. With sham surgery, OB was lower after immobilization in the cast leg than in both the noncast leg (-26%, P = 0.023) and the nonimmobilized control (-34%, P = 0.001), but OB subsequently recovered. With castration, OB was lower after immobilization in the cast leg than in the nonimmobilized control (-34%, P = 0.001), and remained depressed at recovery (-34%, P = 0.001). NKA isoforms did not differ after immobilization or recovery in the sham group. After castration, α2 in the cast leg was ~60% lower than in the noncast leg (P = 0.004) and nonimmobilized control (P = 0.004) and after recovery remained lower than the nonimmobilized control (-42%, P = 0.039). After immobilization, β1 was lower in the cast than the noncast leg (-26%, P = 0.018), with β2 lower in the cast leg than in the noncast leg (-71%, P = 0.004) and nonimmobilized control (-65%, P = 0.012). No differences existed for α1 or α3. Thus, both OB and α2 decreased after immobilization and recovery in the castration group, with α2, β1, and β2 isoform abundances decreased with immobilization compared with the sham group. Therefore, testosterone suppression in rats impaired restoration of immobilization-induced lowered number of functional NKA and α2 isoforms in soleus muscle.NEW & NOTEWORTHY: The Na+,K+-ATPase (NKA) is vital in muscle excitability and function. In rats, immobilization depressed soleus muscle NKA, with declines in [3H]ouabain binding, which was restored after 14 days recovery. After testosterone suppression by castration, immobilization depressed [3H]ouabain binding, depressed α2, β1, and β2 isoforms, and abolished subsequent recovery in [3H]ouabain binding and α2 isoforms. This may have implications for functional recovery for inactive men with lowered testosterone levels, such as in prostate cancer or aging.

Telmisartan use in rats with preexisting osteoporotics bone disorders increases bone microarchitecture alterations via PPARγ

AimsTelmisartan (TEL), an angiotensin II type I receptor blocker and PPARγ partial agonist, has been used for to treat hypertension. It is known that PPARγ activation induces bone loss. Therefore, we evaluate the effects of telmisartan on PPARγ protein expression, biomechanics, density and bone microarchitecture of femurs and lumbar vertebrae in SHR ovariectomized animals, a model of hypertension in which preexisting bone impairment has been demonstrated. Main methodsSHR females (3 months old) were distributed into four groups: sham (S), sham + TEL (ST), OVX (C) and OVX + TEL (CT). TEL (5 mg/kg/day) or vehicle were administered according to the groups. After the protocol, blood pressure was measured and density, microarchitecture and biomechanics of bone were analyzed. Western blotting analysis was performed to evaluate PPARγ protein expression in the bones. Key findingsCastration induced a deleterious effect on mineral density and trabecular parameters, with telmisartan enhancing such effects. Telmisartan increased PPARγ levels, which were at their highest when the treatment was combined with castration. As to biomechanical properties, telmisartan reduced the stiffness in the castration group (CT vs. S or C group), as well as resilience and failure load in ST group (vs. all others groups). SignificanceThese results demonstrated that telmisartan compromised bone density and microarchitecture in animals that shows preexisting osteoporotic bone disorders, probably via mechanisms associated with increased PPARγ. If this translates to humans, a need for greater caution in the use of telmisartan by patients that have preexisting bone problems, as in the postmenopausal period, may be in order.

Changes in the oxidative stress factors and inflammatory proteins following the treatment of BPH-induced dogs with an anti-proliferative agent called tadalafil.

Finding a medical treatment which can combat cell proliferation and relax smooth muscles in canine benign prostatic hyperplasia (BPH) appears to be imperative. This study aimed to evaluate the oxidative stress and inflammatory proteins following the treatment of dogs induced for BPH with an anti-proliferative agent called tadalafil. Twenty-five adult intact male dogs were randomly designated into five groups (n=5): Control group was not induced for BPH and not treated with tadalafil; dogs induced for BPH by testosterone enanthate and estradiol benzoate and treated with tadalafil (5mg/day P.O.); dogs which received tadalafil (5mg/day P.O.); dogs induced for BPH and treated with castration; and dogs induced for BPH. Oxidative stress factors (glutathione peroxidase [GPX], superoxide dismutase [SOD], catalase) and inflammatory proteins (haptoglobin, serum amyloid A [SAA], malondialdehyde [MDA]) were measured in the blood serum for four sequential weeks. Glutathione peroxidase and SOD serum levels declined in dogs in the BPH-induced group compared to those in the control group. Those levels diminished in BPH-induced castrated and tadalafil-treated groups. The changes in the GPX and SOD serum concentrations were not significant between the BPH-induced castrated group and BPH-induced tadalafil-treated group. Moreover, MDA concentration increased slightly in groups with BPH and groups which were castrated. Generally, however, there were no significant differences in the MDA serum concentrations between other groups. Haptoglobin and SAA concentrations increased in BPH-castrated group. Also, the differences in haptoglobin and SAA were not significant between the groups. Tadalafil could not control oxidative stress and inflammatory mediators which happened during BPH in dogs.

Effect of low androgen status on the expression of adenosine A 2A and A 2B receptors in rat penile corpus cavernosum

To investigate whether low androgen status affects erectile function by regulating the expression of adenosine A2A and A2B receptors in rat penile corpus cavernosum. Thirty-six 8-week-old male Sprague-Dawley rats were randomly divided into six groups: sham-operated group (4w-sham, 8w-sham), castration group (4w-cast, 8w-cast) and androgen replacement group (4w-cast+T, 8w-cast+T). The rats in the androgen replacement groups were subcutaneously injected with testosterone propionate (3mg/kg) every other day after castration. The maximum intracavernous pressure/mean arterial pressure (ICPmax/MAP), the expression of A2A , A2B , AKT and eNOS and the concentrations of cAMP and cGMP in the corpus cavernosum were detected at the 4th and 8th weeks after the operation. The serum testosterone level and the ratio of ICPmax/MAP decreased significantly in the castration group as compared to other groups (p<0.01). There was no significant difference in the expression of A2A receptor among groups, while the expression of A2B , AKT and eNOS and the concentrations of cAMP and cGMP in the castration group were significantly lower than in other groups (p<0.01). Low androgen status inhibits the AKT/eNOS/cGMP signalling pathways and the production of cAMP in the corpus cavernosum of castrated rats by down-regulating the expression of A2B receptor, and results in decreased of ICPmax/MAP.

Mechanism of androgen in improving erectile dysfunction in castrated rats by regulating androgen receptor/vascular endothelial growth factor

Objective: To explore the mechanism of androgen in improving erectile dysfunction in castrated rats. Methods: Forty 8-week-old male Sprague-Dawley (SD) rats were randomly divided into 4 groups:normal control group (Group A); castration group (Group B, in which rats were castrated); intervention groups (group C and D), in which rats were treated with different concentrations of testosterone undecanoate orally every day at 10 mg/kg (low dose) and 20 mg/kg (high dose), respectively after being castrated. Animals in group A and B were given 0.9% NS instead. After 8-week treatment, the level of serum testosterone, intra cavernous pressure (ICP) and mean arterial pressure (MAP) were detected, and the expression of androgen receptor (AR)and vascular endothelial growth factor (VEGF) were detected in the penis by Immunohistochemistry and Western blot. Results: The level of serum testosterone was significantly lower in group B [(1.3±0.6) nmol/L] than in group A [(17.1±1.5) nmol/L] (P<0.05).After testosterone supplementation, serum testosterone levels in group C [(8.7±1.2) nmol/L] and group D [(15.5±1.6) nmol/L] were higher than that in group B (all P<0.05). Max ICP/MAP of group C and D were higher than that in group B (all P<0.05). Immunohistochemistry and Western blot showed that the expression levels of AR and VEGF in group B were significantly lower than those in group A, C and D, and group D > group C (all P<0.05). Conclusion: Androgen replacement therapy with testosterone undecanoate can improve the erectile function of castrated rats by protecting the integrity of endothelial cells through AR/VEGF pathway.

Gender-Related Effect of Sodium Dichloroacetate on the Number of Hassall's Corpuscles and RNA NKCC1 Expression in Rat Thymus.

The aim was to investigate the effect of dichloroacetate (DCA) on thymus weight, Hassall's corpuscle number (HCs), and NKCC1 RNA expression in Wistar rats aged 4–5 weeks. They were investigated in the controls and DCA-treated gonad-intact and castrated males and females. The treatment lasted 4 weeks with DCA 200 mg/kg/day. At the end of the experiment, rat thymus was weighted, and its lobe was taken for the expression of NKCC1 RNA determined by the PCR method and of Hassall's corpuscles by immunohistochemistry. DCA caused a thymus weight decrease in DCA-treated gonad-intact rats of both genders as compared with their controls (p < 0.05), and no such impact was found in castrated DCA-treated males and females. DCA caused an increase of the HCs in gonad-intact males (p < 0.05), and no such increase in the DCA-treated gonad-intact females was found. There was gender-related difference in the HCs when comparing DCA-treated gonad-intact males and females: males showed significantly higher HCs (p < 0.05); no gender-related differences were found in the castrated DCA-treated groups. The Slc12a2 gene RNA expression level was found to be significantly decreased only in gonad-intact and in castrated DCA-treated males. The authors discuss the gender-related DCA effects on the thymus.

Serum Estradiol/Testosterone Ratio Affects the Development of Benign Prostate Hyperplasia in Rats

ObjectiveBenign prostatic hyperplasia (BPH) develops more likely with increasing age and changing serum concentrations of circulating estradiol (E2) and/or testosterone (T).MethodsIn this study we explored the relationship between serum E2/T ratio and BPH risk in rats by fitting a mathematical model. A total of 176 male rats were randomized divided into normal control, castrated control, and 20 more groups of castrated animals were administered subcutaneously once daily with one of the four dose levels (0.06, 0.17, 0.5, and 1.5 mg per rat) of T combined with one of the five dose levels (2, 6, 17, 50, and 150μg/kg) of E2 in olive oil by different dose ratio. After 30 days, Serial blood samples were obtained to determine serum T and E2 levels by magnetic bead enzyme‐linked immunosorbent assay. The volume of prostate was measured by water displacement method. The prostate specimens were then fixed, embedded in paraffin, and subjected to hematoxylin and eosin staining for pathological examination. MATLAB software was used to simulate the relationship between prostate/body weight ratio (PBR) and E2/T ratio with a mathematical equation.ResultsCompared with the normal control group, the prostate volume decreased significantly (P &lt; 0.05) in the castrated control group and the treatment groups receiving E2/T dose of 2/0.06 or 6/0.06, while the prostate volume increased significantly (P &lt; 0.05) in the other treatment groups. Compared with the normal control group, the PBR increased statistically significant in the treatment groups receiving E2/T dose of 50/0.5, 150/0.5, 2/1.5, 6/1.5,17/1.5, 50/1.5, or 150/1.5 (P &lt; 0.05). The values of PBR, E2 and T in the treatment groups were significantly higher than those in the control groups. Stepwise regression showed that PBR was a function of E2 and T. PBR=−0.1782 + 0.0081E2 + 0.063T − 0.6 × 10−5E2 2 − 0.28 × 10−3T2, (R2=0.6317)ConclusionE2 /T ratio change may be one of the risk factors for PBR, which is associated with the development of BPH.Support or Funding InformationNational Natural Science Foundation of China (No.: 81772429)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Effect of voluntary exercise on intracellular signalling pathways of angiogenesis in the sciatic nerve of type 1 diabetic castrated male rats.

Impaired angiogenesis in sciatic nerve is a major complication of diabetic neuropathy. Protein kinase B (AKT) and extracellular signal regulated kinase (ERK) signaling pathways play critical roles during capillary-like network formation in the angiogenesis process. Twenty-four adult male Wistar rats (weight: 250-300g) were used in the research. The rats were randomly divided into four groups (n = 6): (1) diabetic (Dia), (2) diabetic + castration (Dia-Cas), (3) diabetic + exercise (Dia-E), and (4) diabetic + castration + exercise (Dia-Cas-E). Type 1 diabetes (T1D) was induced with streptozotocin (50mg/kg). After 6 weeks, sciatic nerve was separated and used for histological evaluation and determination of phosphorylated AKT (P-AKT) and phosphorylated ERK (P-ERK) levels by ELISA method. Glucose levels decreased in the Dia-E group compared to the Dia-Cas group (p < 0.01). In addition, our finding shows that exercise in the Dia-Cas group diminished blood glucose levels compared to the Dia-Cas group but this effect of exercise was not significant. Voluntary exercise in the diabetic castrated group decreased P-AKT protein and increased P-ERK 1/2 protein levels in the sciatic tissue compared to the diabetes group significantly (p < 0.05). Histopathological findings showed that Dia-Cas group with 6-week exercise training significantly raised the number of microvascular density in the sciatic tissue of diabetic rats compared to the diabetic group (p < 0.05). Voluntary exercise in diabetic rats increases angiogenesis in the sciatic nerve. The possible mechanism is the increase of P-ERK 1/2 but not P-AKT levels in the sciatic nerve of T1D rats.

\u201cProstate telocytes change their phenotype in response to castration or testosterone replacement\u201d

Telocytes are CD34-positive cells with a fusiform cell body and long, thin cytoplasmic projections called telopodes. These cells were detected in the stroma of various organs, including the prostate. The prostate is a complex gland capable of undergoing involution due to low testosterone levels; and this condition can be reversed with testosterone replacement. Telocyte function in the mature prostate remains to be dermined, and it is not known whether telocytes can take place in tissue remodeling during prostate involution and regrowth. The present study employed structural, ultrastructural and immunohistochemical methods to investigate the telocyte’s phenotypes in the ventral prostate (VP) from control (CT), castrated (CS) and testosterone replacement (TR) groups of adult male Wistar rats. Telocytes were found in the subepithelial, perimuscular and interstitical regions around glandular acini. Telocytes from CT animals have condensed chromatin and long and thin telopodes. In CS group, telocytes appeared quiescent and exhibited layers of folded up telopodes. After TR, telocytes presented loose chromatin, abundant rough endoplasmic reticulum and enlarged telopodes, closely associated with bundles of collagen fibrils. We called these cells “telocytes with a synthetic phenotype”. As testosterone levels and glandular morphology returned toward to the CT group parameters, after 10 days of TR, these telocytes progressively switched to the normal phenotype. Our results demonstrate that telocytes exhibit phenotypic plasticity upon androgen manipulation and interact with fibroblast and smooth muscle cells to maintain glandular architecture in control animals and during tissue remodeling after hormonal manipulation.

Testosterone preserves endothelial function through regulation of S1P1/Akt/FOXO3a signalling pathway in the rat corpus cavernosum.

The aim of the study was to investigate how testosterone regulating endothelial function in the corpus cavernosum of rats. A total of 32 male Sprague-Dawley (SD) rats, 10-weeks-old, were divided randomly into four groups: normal control group (Control); castration group (Castration); the other 16 rats were castrated followed by testosterone supplementation (orally) every day for 8weeks: castration +10mg/kg (lower dose) testosterone group (Castration +LT) and castration +20mg/kg (high dose) testosterone group (Castration +HT). The data showed that androgen deficiency in the Castration group could induce oxidative stress to attenuate endothelial function, manifested by the impairment of endothelial intercellular junction and endothelial content. This was in parallel with a significant decrease in Akt, Akt target and FOXO3a phosphorylation. Testosterone supplementation in the Castration +LT and Castration +HT groups could greatly preserve testosterone serum levels, endothelial function and erectile function through activation of sphingosine-1-phosphate receptor 1 (S1P1)/Akt/FOXO3a pathway. Together, this study suggested that S1P1/Akt/FOXO3a pathway was involved in endothelial dysfunction in the context of androgen deficiency and oxidative stress, which might further explain the mechanism of androgen deficiency inducing ED.

Effect of low androgen levels on the sulphur dioxide signalling pathway in rat penile corpus cavernosum.

The aim of this study was to investigate the relationship between sulphur dioxide (SO2 ) signalling pathway and the changes in erectile function under low androgen levels. Thirty-six healthy male Sprague Dawley (SD) rats aged eight weeks were randomly divided into androgen replacement group, castration group and sham group. Rats in the androgen replacement group were subcutaneously injected with testosterone propionate at 3mg/kg every other day postcastration. The maximum intracavernous pressure/mean arterial pressure (ICPmax /MAP) and the relative content of SO2 in the penile corpus cavernosum were measured. The mRNA and protein expressions of aspartate aminotransferase (AAT1 and AAT2), cysteine oxidase (CDO), endothelial nitric oxide synthase (eNOS) and phosphorylation of endothelial nitric oxide synthase (P-eNOS) were detected. ICPmax /MAP, P-eNOS/eNOS and the level of SO2 decreased significantly in the castration group compared to the other groups (p<0.05). The expressions of mRNA and protein decreased significantly in the castration group compared to the androgen replacement group and the sham group (p<0.05), while there was no significant difference between the androgen replacement group and sham group. Low androgen levels can inhibit erectile function by downregulating the SO2 signalling pathway.

AB011. The regulatory effects of androgen in wound healing of the prostatic urethra after thulium laser resection of the prostate

BackgroundThis research aims to determine the role of androgen in wound healing of the prostatic urethra after thulium laser resection of the prostate (TmLRP). Further, the change of basal cell proliferation and differentiation as well as macrophage polarization and the inflammatory response influenced by androgen were explored. Moreover, the macrophage cytokines effected by finasteride were detected in urine specimens from thulium laser prostatectomy patients, thus to reveal the mechanism of androgen in wound repair process after prostatectomy.Methods(I) Twenty-four beagles that received TmLRPs were randomly distributed into a castration group, a testosterone undecanoate (TU) group and a control group. The prostate wound was assessed once a week using a cystoscope. Histological analysis was then carried out to study the re-epithelialization of the prostatic urethra in each group; (II) immunohistochemical and immunofluorescence techniques were used to determine the basal cell proliferation and differentiation influenced by androgen; (III) the macrophage polarization and inflammatory response in the wound tissue and urine was also investigated; (IV) the macrophage cytokines effected by finasteride were detected in urine specimens from thulium laser prostatectomy patients by enzyme-linked immunosorbent assay (ELISA).Results(I) The healing of the prostatic urethra after a TmLRP was more rapid in the castration group and slower in the TU group than that in the control group; (II) castration accelerated re-epithelialization by promoting basal cell proliferation in the wound surface and beneath the wound and by accelerating the differentiation of basal cells into urothelial cells; (III) castration reduced the duration of the inflammatory phase and induced the conversion of M1 macrophages to M2 macrophages, thus accelerating the maturation of the wound. By contrast, androgen supplementation enhanced the inflammatory response and prolonged the inflammatory phase. Moreover, the anti-inflammatory phase was delayed and weakened. TNF-α, IL-6 and IL-12 that secreted by M1 were higher than the other two groups, and IL-10 and TGF-1 that secreted by M2 were obviously postponed compared with the other two groups; (IV) in the urine specimens from thulium laser prostatectomy patients, the expression of TNF-α, IL-6 and IL-12 were lower while the expression of IL-10 and TGF-1 were higher under the influence of finasteride.Conclusions(I) Androgen deprivation promotes re-epithelialization of the wound, regulates the inflammatory response, and accelerates wound healing of the prostatic urethra after a TmLRP; (II) castration accelerates re-epithelialization by promoting basal cell proliferation in the wound surface and beneath the wound and by accelerating the differentiation of basal cells into urothelial cells; (III) androgen inhibits the prostatic urethra wound healing by regulating the macrophage polarization; (IV) this research established foundation to reveal the mechanism and application of anti-androgen in the treatment of wound healing after prostatectomy.

Leukocyte coping capacity chemiluminescence as an innovative tool for stress and pain assessment in calves undergoing ring castration.

Pain and stress assessment in animals is considered an imperative issue and also a difficult challenge. Unfortunately, no gold standard technique for pain and stress assessment in animals has been validated nowadays. A new tool to assess stress in animals consists of measuring the leukocyte coping capacity (LCC). The aim of this study was to evaluate the whole-blood LCC chemiluminescence as an innovative tool for stress and pain assessment in the bovine species undergoing ring castration. Twenty 2-mo-old male mix-breed Piemontese-Angus-Belgian Blue calves (Bos taurus) weighing 90 ± 4 kg were used. The animals were randomly allocated in 2 groups composed of 10 subjects each as follows: ring castration group (CAS) and sham castration group (SHAM). Blood drawing, scrotal and perineal temperature recording, scrotal lesion score, pain assessment, and LCC Chemiluminescence were performed at different time points, which were as follows: 1 h before castration/sham (-1 h), 30 min postcastration/sham (30 min), 3 d postcastration/sham (3 d), 7 d postcastration/sham (7 d), 14 d postcastration/sham (14 d). Results showed that in CAS LCC values significantly increased (P < 0.05) at 3 d and decreased at 7 d, whereas in SHAM, LCC values did not significantly vary between the study times. Significant differences in LCC values between CAS and SHAM were seen at 7 d (P < 0.0001). In the CAS group, scrotal lesion was scored as 0, 0, 3.8, 2.7, and 0.2 at -1 h, 30 min, 3 d, 7 d, and 14 d, respectively, whereas in SHAM, its score was 0 at every time point. Perineal temperatures did not vary throughout all the study times in both CAS and SHAM. Differences among the 2 groups were noted in scrotal temperatures only at 3, 7, and 14 d (P < 0.05). In CAS, the percentage of animals which obtained a pain score ≥ 1 was: 10% at -1 h, 30% at 30 min, 20% at 3 and 7 d, and 10% at 14 d, whereas in SHAM, no pain signs were noted at any time point. No significant difference between CAS and SHAM was recorded in cortisol blood level at any time point. No stress leukogram nor variation in neutrophil/lymphocyte ratio was noted at any of the time points in both CAS or SHAM. Our results suggest that ring castration might cause long-lasting pain in calves, but its magnitude is not easily detected by conventional methods. We argue that whole-blood LCC chemiluminescence might be a useful tool for detecting pain and stress in calves undergoing ring castration.