Modulatory effects of Saccharomyces cerevisiae var. boulardii on experimentally induced benign prostatic hyperplasia in rats

Benign prostatic hyperplasia (BPH) is an age-related non-neoplastic disease of the prostate gland in men that has become a global health issue in recent years. Due to the side effects of conventional treatment options, attention is now focused on phytotherapeutics for its management. We investigated the possible protective effect of Saccharomyces cerevisiae var. boulardii in a rat model of testosterone propionate (TP) induced BPH. Rats were divided into five groups: Gr. I, untreated control group; Gr. II, TP group; Gr. III, TP + finasteride; Gr. IV, TP + S. cerevisiae var. boulardii; and Gr. V, S. cerevisiae var. boulardii group. Treatments were given daily for 28 days. At the end of the experiment, all rats were weighed and the prostatic indices, prostate specific antigen, serum testosterone concentration as well as the histological and histomorphometric changes were evaluated. Saccharomyces cerevisiae var. boulardii significantly (P <0.05) reduced prostate weight, prostatic index, serum prostate specific antigen, prostatic epithelial thickness and increased luminal diameter. Thus, the results of this study suggest that S. cerevisiae var. boulardii is a potential pharmacological candidate for management of benign prostatic hyperplasia.

The effect of androgen on wool follicles and keratin production in Hetian sheep / Efeito do androgênio na produção de folículos e queratina em ovinos Hetianos

Abstract To investigate the optimal androgen concentration for culturing Hetian sheep wool follicle and to detect effects of androgen concentration on wool follicle cell proliferation and apoptosis using immunofluorescence labeling and real-time quantitative fluorescence determinations of wool keratin-associated protein gene expression levels. Wool follicles were isolated by microdissection and wool follicles and skin pieces were cultured in various concentrations of dihydrotestosterone (DHT) in culture medium. Next, daily lengthwise growth measurements of wool follicles were obtained using a microscopic micrometer. Cultured Hetian wool follicles were stained using the SACPIC method to reveal wool follicle structure, while sheep skin slices were used to observe cell proliferation by immunostaining and cell apoptosis using the TUNEL method. At the molecular biological level, keratin-associated protein (Kap) gene expression was studied using wool follicles cultured for various numbers of days in vitro. Effects of androgen concentrations on Hetian wool follicle growth and development were experimentally studied. EdU proliferation assays revealed that androgen promoted cell proliferation within wool follicle dermal papillae. TUNEL apoptosis detection demonstrated that androgen treatment could delay cell apoptosis. Quantitative reverse transcription polymerase chain reaction (qPCR) results demonstrated that gene expression level patterns of Hetian mountain sheep super-high sulfur protein. Kap1.1, KIF1.2, Kap2.12 and Kap4.2 gene expression level of the mountainous experimental group was significantly higher than plains Hetian sheep. An androgen concentration of 100 nM can promote the growth of Hetian wool follicle cells in vitro, resulting in overexpression of some genes of the Kap family.

Resumo Investigar a concentração ideal de andrógenos em cultura de folículos pilosos de carneiro Hetiano e detectar os efeitos da concentração de andrógenos na proliferação e apoptose de células foliculares, por meio de imunofluorescência e de determinação quantitativa, em tempo real, da fluorescência dos níveis de expressão gênica de proteína associada à queratina. Folículos pilosos foram isolados por microdissecção, e folículos de lã e pedaços de pele foram cultivados em várias concentrações de di-hidrotestosterona (DHT) em meio de cultura. Em seguida, medições diárias de crescimento longitudinal dos folículos capilares foram obtidas usando um micrômetro microscópico. Folículos de lã cultivados de Hetianos foram corados pelo método SACPIC para revelar a estrutura do folículo piloso, enquanto fatias de pele de carneiro foram usadas para observar a proliferação celular por imunocoloração e apoptose celular por meio do método TUNEL. Em âmbito da biologia molecular, a expressão gênica da proteína associada à queratina (Kap) foi estudada usando folículos capilares cultivados por vários dias, in vitro. Os efeitos das concentrações de andrógenos no crescimento e desenvolvimento dos folículos de lã de Hetianos foram estudados experimentalmente. Ensaios de proliferação de EdU revelaram que o andrógeno promoveu a proliferação celular dentro das papilas dérmicas do folículo piloso. A detecção de apoptose por TUNEL demonstrou que o tratamento com andrógeno poderia atrasar a apoptose celular. Os resultados da reação em cadeia da polimerase transcrição reversa quantitativa (qPCR) demonstraram que os padrões de expressão gênica da proteína de enxofre Kap1.1, KIF1.2, Kap2.12 e Kap4.2 foram significativamente maiores no grupo de ovinos Hetianos de montanha. Uma concentração de androgênio de 100 nM pode promover o crescimento de células foliculares de lã de Hetianos in vitro, resultando na superexpressão de alguns genes da família Kap.

Effects of dihydrotestosterone on the expression of anti-Müllerian hormone in primary follicular granulosa cells in rats / 解剖学报

Objective To explore how dihydrotestosterone (DHT) affects the expression of anti-Müllerian hormone (AMH) in primary rat follicular granulosa cells, and then provides a new experimental basis for the role of DHT and AMH in follicular development. Methods Granulosa cells was extracted from the ovaries of 95 21 days SD female rats for primary culture. Firstly, cell morphology were detected by HE staining and follicke stimulating hormone receptor (FSHR) immunofluorescence detected cell purity after 48 hours of cell culture. Then, cells were randomly divided into control group (without drug intervention), 10

The influence of dihydrotestosterone on cytokine releases in male mice model with Graves disease / 中华内科杂志

Objective@#To study the influences of dihydrotestosterone (DHT) on the development of experimental autoimmune Graves disease (EAGD), and to observe the effect of DHT on cytokines in male BALB/c mice model.@*Methods@#Male BALB/c mice aged 6-8 weeks were divided into 4 groups using random number table: (1) control group; (2) EAGD group; (3) placebo group; (4) DHT group. EAGD mice were induced with an adenovirus expressing the human thyroid stimulating hormone receptor antibody A-subunit (Ad-TSHR289). DHT (5mg) or a matching placebo were implanted one week before the first immunization. Thyroid hormones were detected with radioimmunoassay kit.. Cytokines [such as interferonγ (IFNγ), interleukin (IL)-4, IL-10, IL-9, and IL-17] producing cells from the spleen were detected using flow cytometry.@*Results@#As expected Ad-TSHR289 treatment increased total thyroxine [EAGD group vs. control group: (117.76±32.69) nmol/L vs. (33.08±12.61) nmol/L, P<0.0001] and free thyroxine [EAGD group vs. control group: (15.01±11.55) pmol/L vs. (3.55±1.88) pmol/L, P<0.0001]. Treatment of DHT slightly lowered thyroid hormones [DHT group vs. placebo group: total thyroxine (114.80±44.27) nmol/L vs. (123.17±77.73) nmol/L; free thyroxine (13.48±6.01) pmol/L vs. (14.19±12.65) pmol/L], without significant difference (all P>0.05)]. However, the percentage of IL-10, but not IFN γ, IL-4, IL-9 and IL-17, secreted spleen cells increased in DHT group than in the placebo group [(7.11±3.29)% vs. (3.51±1.36)%, P<0.05].@*Conclusion@#The effects of DHT on thyroid hormone are mild. It might play an immunomodulatory role in the male mouse Graves disease model by up-regulating the cytokine IL-10.

Complete Androgen Insensitivity Syndrome and Literature Review / Síndrome da insensibilidade androgênica completa e revisão de literatura

BACKGROUNG: Complete Androgen Insensitivity Syndrome (CAIS) has been reported since 1923, but in 1953 it became known as "testicular feminization". It is a rare recessive genetic disorder linked to the X chromosome that results in different mutations in the androgen receptor. The main clinical presentation in childhood is the presence of bilateral inguinal hernia in phenotypically female subjects. Incidence of androgen insensitivity syndrome in phenotypically females with inguinal hernia is estimated in 0.8% to 2.4%. This is a case report of complete androgen insensitivity syndrome and literature review of preoperative diagnostic methods. CASE SUMMARY: We present a 3 years and 6 months old child with female phenotype, born in São Paulo, Brazil which was diagnosed intraoperatively with complete androgen insensitivity syndrome, during inguinal hernia repair and present potential diagnostic alternatives that we consider viable options in order to avoid this kind of surprise during surgery. CONCLUSION: Investigation of CAIS should be standard in pre-pubertal girls with bilateral inguinal hernia, genetic techniques involving X chromatin or Y chromosome tests present the best choices.

INTRODUÇÃO: A síndrome da insensibilidade androgênica completa (SIAC) é relatada desde 1923, mas foi em 1953 que ficou conhecida como "feminilização testicular". É uma doença genética recessiva rara, ligada ao cromossomo X, causando diversas mutações no receptor de androgênio. A principal apresentação clínica na infância é a presença de hérnia inguinal bilateral em indivíduos fenotipicamente femininos com uma incidência estimada de 0,8% a 2,4%. Apresentamos um caso de insensibilidade androgênica completa, com revisão de literatura dos métodos diagnósticos pré operatórios. Relato do Caso: Apresentamos uma criança de 3 anos e 6 meses de idade com fenótipo feminino, nascida em São Paulo, Brasil diagnosticada com síndrome da insensibilidade androgênica completa, durante a cirurgia de herniorrafia inguinal bilateral e apresentamos potenciais alternativas diagnósticas a fim de evitar esse tipo de surpresa durante a cirurgia. CONCLUSÃO: Em meninas pré-puberes, portadoras de hérnia inguinal bilateral, a pesquisa de SIAC se faz necessária, técnicas genéticas que utilizam a pesquisa da cromatina X ou do cromossomo Y seriam as melhores escolhas.

Identification of three novel SRD5A2 mutations in Chinese patients with 5α-reductase 2 deficiency / 亚洲男科学杂志(英文版)

In this study, we investigated the genetics, clinical features, and therapeutic approach of 14 patients with 5α-reductase deficiency in China. Genotyping analysis was performed by direct sequencing of PCR products of the steroid 5α-reductase type 2 gene (SRD5A2). The 5α-reductase activities of three novel mutations were investigated by mutagenesis and an in vitro transfection assay. Most patients presented with a microphallus, variable degrees of hypospadias, and cryptorchidism. Eight of 14 patients (57.1%) were initially reared as females and changed their social gender from female to male after puberty. Nine mutations were identified in the 14 patients. p.G203S, p.Q6X, and p.R227Q were the most prevalent mutations. Three mutations (p.K35N, p.H162P, and p.Y136X) have not been reported previously. The nonsense mutation p.Y136X abolished enzymatic activity, whereas p.K35N and p.H162P retained partial enzymatic activity. Topical administration of dihydrotestosterone during infancy or early childhood combined with hypospadia repair surgery had good therapeutic results. In conclusion, we expand the mutation profile of SRD5A2 in the Chinese population. A rational clinical approach to this disorder requires early and accurate diagnosis, especially genetic diagnosis.

Study on effect of extract from Tibetan medicine Urtica hyperborean on anti-prostatic hyperplasia / 中国中药杂志

In this study,mouse models of benign prostatic hyperplasia induced by subcutaneous injection of testosterone propionate was used to investigate the therapeutic effect and mechanism of Urtica hyperborean( UW) extracts on prostate hyperplasia in mice. The effects of UW extracts on prostate index,serum epidermal growth factor( EGF) and dihydrotestosterone( DHT) in model mice were observed,and the EGF and anti-apoptotic factor( Bcl-2) mRNA expression levels were detected as well as pathological changes in prostate tissue. The results showed that the ethyl acetate extraction and alcohol soluble fraction of the UW could significantly reduce the prostate index,reduce the serum DHT and EGF levels( P<0. 01),and significantly decrease the EGF and Bcl-2 mRNA expression( P<0. 01),significantly improved the morphological structure of prostate tissue. The above results confirmed that ethyl acetate extract and alcohol-soluble parts of UW have a good preventive effect on mice prostatic hyperplasia model,and its mechanism may be to reduce androgen levels by regulating polypeptide growth factors and/or inhibiting cell hyperproliferation and promoting apoptosis. This study laid the foundation for the further research on UW.

Identification of three novel SRD5A2 mutations in Chinese patients with 5α-reductase 2 deficiency / 亚洲男科学杂志(英文版)

In this study, we investigated the genetics, clinical features, and therapeutic approach of 14 patients with 5α-reductase deficiency in China. Genotyping analysis was performed by direct sequencing of PCR products of the steroid 5α-reductase type 2 gene (SRD5A2). The 5α-reductase activities of three novel mutations were investigated by mutagenesis and an in vitro transfection assay. Most patients presented with a microphallus, variable degrees of hypospadias, and cryptorchidism. Eight of 14 patients (57.1%) were initially reared as females and changed their social gender from female to male after puberty. Nine mutations were identified in the 14 patients. p.G203S, p.Q6X, and p.R227Q were the most prevalent mutations. Three mutations (p.K35N, p.H162P, and p.Y136X) have not been reported previously. The nonsense mutation p.Y136X abolished enzymatic activity, whereas p.K35N and p.H162P retained partial enzymatic activity. Topical administration of dihydrotestosterone during infancy or early childhood combined with hypospadia repair surgery had good therapeutic results. In conclusion, we expand the mutation profile of SRD5A2 in the Chinese population. A rational clinical approach to this disorder requires early and accurate diagnosis, especially genetic diagnosis.

RE-ORGA, a Korean Herb Extract, Can Prevent Hair Loss Induced by Dihydrotestosterone in Human Dermal Papilla Cells

BACKGROUND: Androgenic alopecia (AGA) is the most common type of hair loss. It is likely inherited genetically and is promoted by dihydrotestosterone. 5α-reductase has been proven a good target through finasteride use. However, the pathogenesis of AGA cannot be fully explained based only on dihydrotestosterone levels. OBJECTIVE: To identify similar hairloss inhibition activity of RE-ORGA with mode of action other than finasteride. METHODS: We prepared RE-ORGA from Korean herb mixtures. We performed MTT assays for cytotoxicity, Cell Counting Kit-8 assays for cell proliferation, and western blot to identify expression levels of 5α-reductase and Bax. RNA-sequencing was performed for the expression patterns of genes in dihydrotestosterone-activated pathways. Anti-inflammatory activity was also assessed by the expression levels of tumor necrosis factor-alpha (TNF-α) and interleukin 6. RESULTS: REORGA could promote the proliferation of human dermal papilla cells and showed low cytotoxicity. It also inhibited the expression of 5α-reductases and Bax in the cells. RNA-sequencing results verified that the mRNA expressions of SRD5A1, Bax, transforming growth factor-beta 1 (TGF-β1), and TGF-β1 induced transcript 1 (TGFβ1I1) were decreased, whereas expression of protein tyrosine kinase 2 beta (PTK2β) was more elevated. REORGA also showed anti-inflammatory activity through decreased mRNA levels of TNF-α. CONCLUSION: Transcriptionally, up-regulation of PTK2β and concomitant down-regulation of TGFβ1I1 imply that RE-ORGA can modulate androgen receptor sensitivity, decreasing the expression of 5α-reductase type II and Bax together with TGF-β1 transcripts; RE-ORGA also showed partial anti-inflammatory activity. Overall, RE-ORGA is expected to alleviate hair loss by regulating 5α-reductase activity and the receptor's androgen sensitivity.

Inhibitory effects of Pycnogenol®, a pine bark extract, in a rat model of testosterone propionate-induced benign prostatic hyperplasia / 한국실험동물학회지

Benign prostate hyperplasia (BPH) is a male reproductive disease that has gained increasing importance in recent years. The present study investigated whether Pycnogenol® (PYC), a standardized French maritime pine bark extract, could prevent BPH induced by testosterone propionate (TP) in rats. Male Sprague-Dawley rats were randomly divided into five groups of six rats. One group was used as a normal control rats and the other groups received subcutaneous injections of TP for 4 weeks to induce BPH. In the two treatment groups, PYC (20 or 40 mg/kg) was administered daily for 4 weeks by oral gavage concurrently with the induction of TP. All rats were sacrificed at the scheduled termination time, the prostates were weighed, and histopathologic examinations were conducted. Dihydrotestosterone (DHT) levels in serum and the prostate were measured, and the expression of proliferating cell nuclear antigen (PCNA) and Ki-67 proteins was investigated. BPH-treated animals showed increases in the relative weight of the prostate, higher concentrations of DHT in serum and the prostate, and higher expression of PCNA and Ki-67 in the prostate; in contrast, PYC-treated animals had significant reductions in these factors compared with the BPH animals. These findings indicated that PYC inhibited the development of BPH and that this was closely associated with a reduction in DHT concentration.

Testosterone and its metabolites: Differential associations with cardiovascular and cerebrovascular events in men / 亚洲男科学杂志(英文版)

As men grow older, circulating testosterone declines while the incidence of cardiovascular disease increases. Thus, the role of sex hormones as biomarkers, and possibly contributing factors to clinical manifestations of cardiovascular disease in the increasing demographic of aging men, has attracted considerable interest. This review focuses on observational studies of endogenous androgens, namely circulating testosterone and dihydrotestosterone, which have examined their associations with cardiovascular events such as myocardial infarction and stroke. Studies which have examined the associations of endogenous estrogens, namely circulating estradiol, with these outcomes are also discussed. In large prospective cohort studies of predominantly middle-aged and older men, lower circulating testosterone consistently predicts higher incidence of cardiovascular events. Of note, both lower circulating testosterone and lower dihydrotestosterone are associated with higher incidence of stroke. These associations are less apparent when myocardial infarction is considered as the outcome. Results for estradiol are inconsistent. Lower circulating testosterone has been shown to predict higher cardiovascular disease-related mortality, as has lower circulating dihydrotestosterone. It is possible that the relationship of circulating androgens to cardiovascular events or mortality outcomes may be U-shaped rather than linear, with an optimal range defining men at lowest risk. Epidemiological studies are observational in nature and do not prove causality. Associations observed in studies of endogenous androgens need not necessarily translate into similar effects of exogenous androgens. Rigorous randomized controlled trials are needed to clarify the effects of testosterone treatment on cardiovascular risk in men.

Inhibitory Effect of Pueraria Isoflavones on Prostatic Hyperplasia in Rats / 中国药学杂志

OBJECTIVE: To investigate the inhibitory effect and possible mechanisms of puerariae isoflavone(PI) on prostatic hyperplasia induced by testosterone propionate.METHODS: Forty-eight male Wistar rats were randomly divided into six groups according to their body weight including normal control group, model group, 40, 80, 160 mg•kg-1•d-1 PI group, and finasteride positive control group. In addition to the sham operation for rats in the normal control group, the rats in other five groups performed castration surgery. After the restoration, the five groups of rats were subcutaneously injected with testosterone propionate (10 mg•kg-1•d-1) for 10 d to establish a benign prostatic hyperplasia model and then the subcutaneous injection was maintained every 2 d. High, middle and low dose PI groups were intragastrically administered (40, 80, 160 mg•kg-1•d-1) from the second day when the benign prostatic hyperplasia model was successfully constructed. The positive control group was given finasteride (1.0 mg•kg-1•d-1).Rats in normal and model groups were given an equal volume of saline for 28 d. After the last administration, the prostate and seminal vesicles were separated under anesthesia in rats, the wet weight and volume of the prostate and seminal vesicles were measured. The prostate and seminal vesicles index were calculated too. Rat blood was drawn and dihydrotestosterone(DHT) and estradiol (E2) in the serum were measured. Nitric oxide (NO), nitric oxide synthase (NOS), superoxide dismutase (SOD) and malondialdehyde (MDA) levels in prostate tissues were measured. The prostate tissue in each group was randomly selected for HE staining. The pathological structure of the prostate tissue was observed under an optical microscope.RESULTS: Compared with the normal control group, the prostate gland index and seminal vesicle gland index of the model group increased significantly (P<0.01), and the DHT and E2 levels in serum increased significantly (P<0.01). MDA content was increased while NO levels, NOS and SOD activities were significantly decreased (P<0.01). HE staining showed that the size of the prostate gland in the model group was different, there were obvious dilation, hyperplasia and papillary protrusions, and the cavity was full of pink and homogeneous density. The interstitial tissue showed obvious dilations of blood vessels, infiltration of inflammatory cells, and proliferation of fibrous connective tissues. Compared with the model group, the index and volume of prostate and seminal vesicles in the PI and positive control groups were significantly decreased (P<0.05 or P<0.01), and the levels of serum DHT and E2 in the middle and high doses PI groups were significantly lower (P<0.05 or P<0.01). In all treatment groups, MDA content was decreased and NO, NOS, and SOD levels were increased (P<0.05 or P<0.01) except the low-dose PI groups. There was moderately hyperplasia in low-dose PI group, mild prostatic hyperplasia in positive control group and middle-dose PI group, basically no hyperplasia in high-dose PI group.CONCLUSION: PI has a certain inhibitory effect on prostate hyperplasia induced by testosterone propionate, especially in the medium and high dose PI groups. The mechanism may be related to the effects of pueraria isoflavone on antioxidant,free radical scavenging in vivo, increasing NOS activity and increasing NO level.

Establishment of a model of dihydrotestosterone-induced polycystic ovary syndrome complicated with atherosclerosis in rats / 中国基层医药

Objective@#To establish a rat model of polycystic ovary syndrome(PCOS) complicated with atherosclerosis(AS).@*Methods@#Sixteen female SD rats were selected and randomly divided into control group and model group, with 8 rats in each group.The rats in the control group were given routine rearing.The rats in the model group were subcutaneously given dihydrotestosterone(DHT) in neck and fed with high fat diet for a long term.The changes of food intake (FI), body weight(BW), testosterone (T), estrogen (E2), luteinizing hormone (LH), total cholesterol (TC), blood glucose (BG) and insulin (INS) were observed in the two groups.@*Results@#The levels of FI, T, E2, LH between the control group and model group had no obvious change[(86.13±7.83)g/r vs.(96.25±10.66)g/r, t=2.113, P=0.563, (10.79±1.74)mg/L vs.(11.47±1.89)mg/L, t=1.785, P=0.087; (36.58±2.57)ng/L vs.(38.64±1.78)ng/L, t=2.697, P=0.068; (15.47±1.96)IU/L vs.(16.01±0.80)IU/L, t=1.570, P=0.614]. The levels of BW, TC, DHT, INS in the model group were significantly higher than those in the control group[(234.54±17.14)g vs.(192.67±16.47)g, t=7.930, P<0.000; (2.47±0.13)mmol/L vs.(2.02±0.15)mmol/L, t=6.475, P<0.000; (139.75±12.12)ng/L vs.(55.63±7.80)ng/L, t=8.697, P<0.000; (283.25±33.47)pg/mg vs.(162.12±15.51)pg/mg, t=9.289, P<0.000]. Tube wall was markedly thickened in the model group after being given high fat feed for 12 weeks, and intimal wall was significantly thickened after 16 weeks, and endotheliocyte injury, deep collagen fiber, monocyte adhesion, visible atherosclerotic plaques were observed in the model grouop.@*Conclusion@#DHT-induced PCOS rat model by high fat feed reproduces the human typical clinical symptoms and pathological characteristics, it can provide a relatively simple and feasible rat model for further study of the mechanism of PCOS complicated with AS.

Correlations between DHT/AR and ATAD2 expression in hepatocellular carcinoma / 中华肝胆外科杂志

Objective To investigate the expression of androgen receptor(AR),ATAD2 in hepatocellular carcinoma(HCC) and the correlations with clinicopathological features,and the role of DHT/AR and ATAD2 in proliferation of HCC cells.Methods The samples of 75 patients with HCC in the First Affiliated Hospital of China Medical University from February 2012 to December 2012 were collected.LM3 and Huh7 cells were divided into control group,DHT group,DHT + CDX (bicalutamide) group and CDX group;and also divided into Ri-ATAD2 group (adding interference fragments) and Ri-C group (adding control vector sequence).Immunohistochemistry was used to detect the expression of AR and ATAD2,and to analyze the correlations between clinical features and survival of patients.Real-time PCR and Western Blot were used to detect the expression of AR and ATAD2,and CCK-8 was used to detect cell proliferation.Results HCC patient samples were grouped according to AR and ATAD2 expression.Compared with low AR expression group (n =31),the ratio of tumor ＜5 cm in high expression group (n =44) was higher,and the ratio of TNM stage Ⅰ + Ⅱ was lower.Compared with low ATAD2 expression group (n=35),the ratio of metastasis and tumor differentiation grade Ⅲ + Ⅳ was higher in high expression group (n=40),and the difference was statistically significant (P ＜ 0.05).The overall survival rate of patients with high expression of ATAD2 was lower than other patients,and the differences were statistically significant (P＜0.05).Multivariate Cox regression analysis showed that ATAD2 expression (HR=1.935,95％ CI:1.066～3.515) and metastasis (HR=2.212,95％ CI:1.059～4.619) were independent predictors of poor prognosis.Compared with LO2 cells,the mRNA and protein level of AR and ATAD2 in LM3 and Huh7 cells were significantly higher,and the differences were statistically significant (P＜0.05).And the proliferation rate of HCC cells increased significantly after 48 and 72 hours compared with the control group,and the differences were statistically significant (P＜0.05).After adding CDX,the proliferation of LM3 and Huh7 induced by DHT was inhibited.DHT enhanced the expression of ATAD2,while CDX inhibited the expression of ATAD2.The expression of ATAD2 protein decreased when LM3 and Huh7 cells were interfered.Compared with Ri-C group,the proliferation of HCC cells in Ri-ATAD2 group decreased significantly after the DHT treatment 48 and 72 hours,and the difference was statistically significant (P＜0.05).Conclusions DHT/AR promoted the proliferation of HCC cells by inducing ATAD2 expression.Modulating ATAD2 expression may be the potential mechanism of DHT/AR in HCC proliferation.

Testosterone and its metabolites: differential associations with cardiovascular and cerebrovascular events in men / 亚洲男科学杂志(英文版)

As men grow older, circulating testosterone declines while the incidence of cardiovascular disease increases. Thus, the role of sex hormones as biomarkers, and possibly contributing factors to clinical manifestations of cardiovascular disease in the increasing demographic of aging men, has attracted considerable interest. This review focuses on observational studies of endogenous androgens, namely circulating testosterone and dihydrotestosterone, which have examined their associations with cardiovascular events such as myocardial infarction and stroke. Studies which have examined the associations of endogenous estrogens, namely circulating estradiol, with these outcomes are also discussed. In large prospective cohort studies of predominantly middle-aged and older men, lower circulating testosterone consistently predicts higher incidence of cardiovascular events. Of note, both lower circulating testosterone and lower dihydrotestosterone are associated with higher incidence of stroke. These associations are less apparent when myocardial infarction is considered as the outcome. Results for estradiol are inconsistent. Lower circulating testosterone has been shown to predict higher cardiovascular disease-related mortality, as has lower circulating dihydrotestosterone. It is possible that the relationship of circulating androgens to cardiovascular events or mortality outcomes may be U-shaped rather than linear, with an optimal range defining men at lowest risk. Epidemiological studies are observational in nature and do not prove causality. Associations observed in studies of endogenous androgens need not necessarily translate into similar effects of exogenous androgens. Rigorous randomized controlled trials are needed to clarify the effects of testosterone treatment on cardiovascular risk in men.

Corni Fructus attenuates testosterone-induced benign prostatic hyperplasia by suppressing 5α-reductase and androgen receptor expression in rats

BACKGROUND/OBJECTIVES: Benign prostatic hypertrophy (BPH) is a major cause of abnormal overgrowth of the prostate mainly in the elderly. Corni Fructus has been reported to be effective in the prevention and treatment of various diseases because of its strong antioxidant effect, but its efficacy against BPH is not yet known. This study was designed to evaluate the therapeutic efficacy of Corni Fructus water extract (CF) in testosterone-induced BPH rats. MATERIALS/METHODS: To induce BPH, rats were intraperitoneal injected with testosterone propionate (TP). Rats in the treatment group were orally administered with CF with TP injection, and finasteride, which is a selective inhibitor of 5α-reductase type 2, was used as a positive control. RESULTS: Our results showed that the increased prostate weight and histopathological changes in BPH model rats were suppressed by CF treatment. CF, similar to the finasteride-treated group, decreased the levels of testosterone and dihydrotestosterone by TP treatment in the serum, and it also reduced 5α-reductase expression and concentration in prostate tissue and serum, respectively. In addition, CF significantly blocked the expression of the androgen receptor (AR), AR co-activators, and proliferating cell nuclear antigen in BPH rats, and this blocking was associated with a decrease in prostate-specific antigen levels in serum and prostate tissue. CONCLUSIONS: These results suggest that CF may weaken the BPH status through the inactivation of at least 5α-reductase and AR activity and may be useful for the clinical treatment of BPH.

Study of the Biological Activities Preparation with Tribulus Terrestris l. on Free Serum Testosterone Changes in Male Rats / Монголын эм зүй, эм судлал

Wistar nonlinear rats weighing 170-220g. Rats were divided 5 groups, including control group, group-1, group-2, group -3 and reference group. Dried thistle extract and raw bovine testicle were contained by 1:1, 1:2 and 2:1 ratio. Each 0.1g ratio was dissolved in 20 ml distilled water and administered 2 times per day. Blood sampling was done for each rat after 3, 7, 14, 21, 28 and 35 days. Their testosterone level was measured by ELISA Kit. Results: The results indicated that free serum testosterone level in male rats increase and decrease in 7 days frequency. All tested groups showed gradual increase in the level of free serum testosterone when compared to that of corresponding control (p<0.05). Statistical comparison of all groups revealed that the maximum level was found in group 1. Moreover, group 3 was showed gradually increase in level of free serum testosterone, irrelative with period of decrease testosterone level. Conclusion: According to our results and previous study, it is suggested that preparation with Tribulus terrestris L. extract could be used in the androgen deficiency and erectile dysfunctions. Keywords: Tribulus Terrestris L, Free testosterone, Dihydrotestosterone, Protodioscin

Enhanced transdermal delivery of dihydrotestosterone using the combination strategy of iontophoresis and prodrug / 国际药学研究杂志

Objective To enhance the transdermal delivery of dihydrotestosterone (DHT) and to make the delivery controllable, which would enable personalized administration of DHT to pediatric patients. Methods A hydrosoluble, ionizable prodrug of DHT, dihydrotestosterone dimethylglycine ester hydrochloride (DHT-DG), was synthesized, characterized and its physicochemical properties were determined. DHT And DHT-DG were formulated into hydrogel, respectively, and the transdermal delivery of each compound across fresh porcine ear skin with or without iontophoresis was evaluated. Results Application of current (0.5 mA/cm2) to 2.5% DHT-DG hydrogel for 8 h enabled (226.91 ± 45.62) nmol/cm2 accumulative amounts of DHT-DG across fresh porcine ear skin, which was much higher than the amount of DHT delivered from 2.5% DHT hydrogel without iontophoresis, (10.45±3.63) nmol/cm2. More than 80% delivered amount of DHT-DG was hydrolyzed into its parent drug DHT. The steady state flux of DHT-DG was found after the application of iontophoresis for 2 h, and the accumulative amounts of DHT-DG could be modulated by drug concentration and current intensity. Conclusion Combination strategy of iontophoresis and prodrug can enable fast controllable transdermal delivery of DHT.

Enhanced transdermal delivery of dihydrotestosterone using the combination strategy of iontophoresis and prodrug / 国际药学研究杂志

Objective To enhance the transdermal delivery of dihydrotestosterone(DHT)and to make the delivery controlla?ble,which would enable personalized administration of DHT to pediatric patients. Methods A hydrosoluble,ionizable prodrug of DHT,dihydrotestosterone dimethylglycine ester hydrochloride(DHT-DG),was synthesized,characterized and its physicochemical properties were determined. DHT And DHT-DG were formulated into hydrogel,respectively,and the transdermal delivery of each compound across fresh porcine ear skin with or without iontophoresis was evaluated. Results Application of current(0.5 mA/cm2)to 2.5%DHT-DG hydrogel for 8 h enabled(226.91±45.62)nmol/cm2 accumulative amounts of DHT-DG across fresh porcine ear skin, which was much higher than the amount of DHT delivered from 2.5%DHT hydrogel without iontophoresis,(10.45±3.63)nmol/cm2. More than 80%delivered amount of DHT-DG was hydrolyzed into its parent drug DHT. The steady state flux of DHT-DG was found af?ter the application of iontophoresis for 2 h,and the accumulative amounts of DHT-DG could be modulated by drug concentration and current intensity. Conclusion Combination strategy of iontophoresis and prodrug can enable fast controllable transdermal delivery of DHT.

Androgen receptor up-regulates EphA3 expression in androgen-dependent prostate cancer cell lines / 天津医药

Objective To evaluate the relationship between liver cell type A receptor (EphA) expression and androgen receptor (AR) signaling in androgen-dependent prostate cancer cells. Methods RT-PCR and Western blot assay were used to determine mRNA and protein levels of EphA3 and AR in prostate cancer LNCaP and 22Rv1 cells, respectively. The variations of EphA3, AR and prostate specific antigen (PSA) expressions were also measured in these cells after dihydrotes?tosterone (DHT) treatment for 48 h. The constructed EphA3-Luc (-789-+146) luciferase reporter plasmid was co-transfect?ed with pcDNA3.1(+)-AR or siAR in 22Rv1 cells to analyze the effects of different AR expression levels on EphA3 tran?scription activity. Results The expression pattern of EphA3 was similar to AR, showing a lower level in prostate stromal cell line WPMY-1 and a higher level in prostate cancer cell lines LNCaP and 22Rv1. When stimulated with 10 nmol/L DHT, the expression levels of AR, PSA and EphA3 were significantly increased in 22Rv1 cells, and the protein levels of these genes were also increased in LNCaP cells. Moreover, AR expression levels markedly influenced the activity of EphA 3 pro?moter. Conclusion AR up-regulates EphA3 expression by increasing the activity of EphA3 promoter.