Preventive effect of Pueraria mirifica on testosterone‐induced prostatic hyperplasia in Sprague Dawley rats

Pueraria mirifica (PM) extract contains phytoestrogen daidzein and genistein. In this study, we investigated the protective effect of PM extract, daidzein and genistein on a testosterone‐induced prostatic hyperplasia in rats. Testosterone was administered at 3 mg kg^?1 to rats followed by the PM extract, daidzein and genistein for a period of 30 days with finasteride as positive control. The testosterone level was increased, indicating inhibition of 5α‐reductase converting testosterone to dihydrotestosterone. This was confirmed by prostate‐specific antigen level that significantly decreased when treated with PM extract, daidzein and genistein. The PM extract, daidzein and genistein reduced the increase in the prostate/body weight ratio in testosterone‐induced rats. This gives indication that PM extract, daidzein and genistein possessed protective activity for the treatment of benign prostatic hyperplasia. The analysis of histoarchitechture of the prostate has also shown that there was a significant improvement in prostatic cells of the testosterone‐induced rats when treated with PM extract, daidzein and genistein.     

Comparison of the response to treatment between Asian and Caucasian men with benign prostatic hyperplasia: Long‐term results from the combination of dutasteride and tamsulosin study

The Combination of Avodart and Tamsulosin study was a 4‐year, randomized, double‐blind study of the efficacy and safety of dutasteride and tamsulosin, alone or in combination, in men with moderate‐to‐severe benign prostatic hyperplasia. In this post‐hoc investigation, we analyzed primary and secondary end‐points from the Combination of Avodart and Tamsulosin study in Asian (n = 325) and Caucasian men (n = 4259). The incidence of acute urinary retention or benign prostatic hyperplasia‐related surgery did not differ significantly between treatment groups in the Asian subpopulation. In Caucasian men, the incidence of acute urinary retention/benign prostatic hyperplasia‐related surgery was significantly lower in the combination therapy group compared with the tamsulosin monotherapy group (P < 0.001), but not compared with dutasteride monotherapy. Combination therapy significantly increased the time to benign prostatic hyperplasia clinical progression and resulted in improved International Prostate Symptom Score, maximum urinary flow rate, quality of life, and reduced prostate volume in Asian and Caucasian men who received combination therapy compared with tamsulosin monotherapy. Combination therapy also significantly improved (P < 0.05) time to benign prostatic hyperplasia clinical progression, International Prostate Symptom Score, maximum urinary flow rate and quality of life versus dutasteride in the Caucasian subpopulation. The adverse‐event profile was comparable between subpopulations. In conclusion, Asian and Caucasian men respond similarly to these treatments, despite apparent racial differences in 5α‐reductase activity.     

NAD(P)H‐quinone oxidoreductase 1 silencing aggravates hormone‐induced prostatic hyperplasia in mice

NAD(P)H‐quinone oxidoreductase 1 (NQO1) is a highly inducible flavoprotein known to involve in various cellular defence mechanisms. In this study, we explored whether NQO1 deletion affects hormone‐induced prostatic hyperplasia. Testosterone propionate (3 mg/kg, IP) was injected into wild‐type (WT) and NOQ1 knockout C57BL/6 mice (NQO1^?/?) for 14 consecutive days, and the samples were collected for biological and histochemical studies. The testosterone‐treated NQO1^?/? showed about 140% higher prostate weight than the testosterone‐treated WT, with enhanced connective tissue and hyperplastic glands formations. However, increased dihydrotestosterone level after testosterone treatment was not significantly different between the WT and NQO1^?/?. In contrast, the enhanced nuclear expression of proliferating cell nuclear antigen in NQO1^?/? prostate confirmed aggravated prostatic hyperplasia in NQO1^?/?. Moreover, the expression of heat shock protein (HSP) 90‐α was markedly increased in the NQO1^?/?, and this was supported by increased testosterone‐induced nuclear androgen receptor expression in NQO1‐silenced LNCaP cells. Testosterone‐induced prostate‐specific antigen expression was not reversed in NOQ1‐silenced cells after finasteride treatment. Although the exact role of NQO1 in prostatic hyperplasia remains unclear, the hyperplasia exacerbation due to NQO1 deletion might be independent of type 2 5α‐reductase and might be related to enhanced androgen receptor affinity due to enhanced HSP90‐α expression.     

Expression of human β‐defensin‐2 in the prostate

What’s known on the subject? and What does the study add? We found the expression of human β‐defensin‐2 (HBD‐2) in the prostate for the first time and LPS, a gram negative bacterial component, upregulated HBD‐2 in prostate epithelial cells. We are looking for other antimicrobial peptides expressed in the prostate besides human β‐defensin‐2. Also, we are studying the relationship between antimicrobial peptides and the development or progression of prostate diseases.

OBJECTIVE

To investigate the expression and regulation of human β‐defensin‐2 (HBD‐2) in the prostate.

PATIENTS AND METHODS

Normal human prostate epithelial cell line (RWPE‐1), human prostate cancer cell lines (DU‐145, PC‐3), and paraffin‐embedded prostate tissue from patients with benign prostatic hyperplasia (BPH) were analysed by RT‐PCR and immunohistochemical staining. HBD‐2 expression was also analysed by RT‐PCR and ELISA in RWPE‐1 cells treated with lipopolysaccharide (LPS). Nuclear factor‐κB (NF‐κB) activation was assessed by IκBα immunoblotting and electrophoretic mobility shift assay (EMSA).

RESULTS

BPH tissue and all of the tested prostate cell lines other than PC‐3 constitutively express HBD‐2 mRNA. HBD‐2 protein was strongly detected in prostate gland tissue surrounded by inflammatory cells including macrophages. Exposure to LPS induced HBD‐2 upregulation and NF‐κB activation, as assessed by IκBα phosphorylation and degradation in RWPE‐1 cells. Bay11‐7082, an NF‐κB inhibitor prevented LPS‐induced HBD‐2 production in RWPE‐1 cells.

CONCLUSIONS

Prostate epithelial cells may constitutively express HBD‐2, and its expression was upregulated by LPS. Our data indicate that HBD‐2 may be an important immunomodulatory factor in prostate function. Expression of HBD‐2 in normal prostates and the potential role of HBD‐2 in prostatitis and BPH should be addressed in the future.     

Effects of long‐term heat stress and dietary restriction on the expression of genes of steroidogenic pathway and small heat‐shock proteins in rat testicular tissue

The aim was to investigate the effects of long‐term heat stress and dietary restriction on the expression of certain genes involving in steroidogenic pathway and small heat‐shock proteins (sHSPs) in rat testis. Sprague Dawley rats (n = 24) were equally divided into four groups. Group I and II were kept at an ambient temperature of 22°C, while Groups III and IV were reared at 38°C for 9 weeks. Feed was freely available for Group I and Group III, while Group II and Group IV were fed 60% of the diet consumed by their ad libitum counterparts. At the end of 9 weeks, testicles were collected under euthanasia. Total RNA was isolated from testis tissue samples. Expression profiles of the genes encoding androgen‐binding protein, follicle‐stimulating hormone receptor, androgen receptor, luteinising hormone receptor, steroidogenic acute regulatory protein (StAR), cyclooxygenase‐2 and sHSP genes were assessed at mRNA levels using qPCR. Long‐term heat stress decreased the expression of StAR and HspB10 genes while dietary restriction upregulated StAR gene expression. The results suggested that long‐term heat stress negatively affected the expression of StAR and HspB10 genes and the dietary restriction was able to reverse negative effect of heat stress on the expression of StAR gene in rat testis.     

High‐fat high‐sucrose diet leads to dynamic structural and inflammatory alterations in the rat vastus lateralis muscle

The influence of obesity on muscle integrity is not well understood. The purpose of this study was to quantify structural and molecular changes in the rat vastus lateralis (VL) muscle as a function of a 12‐week obesity induction period and a subsequent adaptation period (additional 16‐weeks). Male Sprague–Dawley rats consumed a high‐fat, high‐sucrose (DIO, n = 40) diet, or a chow control‐diet (n = 14). At 12‐weeks, DIO rats were grouped as prone (DIO‐P, top 33% of weight change) or resistant (DIO‐R, bottom 33%). Animals were euthanized at 12‐ or 28‐weeks on the diet. At sacrifice, body composition was determined and VL muscles were collected. Intramuscular fat, fibrosis, and CD68+ cells were quantified histologically and relevant molecular markers were evaluated using RT‐qPCR. At 12‐ and 28‐weeks post‐obesity induction, DIO‐P rats had more mass and body fat than DIO‐R and chow rats (p < 0.05). DIO‐P and DIO‐R rats had similar losses in muscle mass, which were greater than those in chow rats (p < 0.05). mRNA levels for MAFbx/atrogin‐1 were reduced in DIO‐P and DIO‐R rats at 12‐ and 28‐weeks compared to chow rats (p < 0.05), while expression of MuRF1 was similar to chow values. DIO‐P rats demonstrated increased mRNA levels for pro‐inflammatory mediators, inflammatory cells, and fibrosis compared to DIO‐R and chow animals, despite having similar levels of intramuscular fat. The down‐regulation of MAFbx/atrogin‐1 may suggest onset of degenerative changes in VL muscle integrity of obese rats. DIO‐R animals exhibited fewer inflammatory changes compared to DIO‐P animals, suggesting a protective effect of obesity resistance on local inflammation. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:2069–2078, 2016.     

Diagnostic significance of miR‐21, miR‐141, miR‐18a and miR‐221 as novel biomarkers in prostate cancer among Egyptian patients

Prostate cancer (PC) is considered as the fifth cause of cancer deaths worldwide. The exact etiopathogenesis is unclear; however, genetic predisposition, hormonal influencers, lifestyle and environmental factors act as major contributors. It has been found that several miRNAs may play a crucial role in cancer initiation and progression. Here, in this study, we evaluated the peripheral blood levels of miR‐21, miR‐141, miR‐221 and miR‐18a expression among 80 prostate cancer patients (50 localised and 30 metastatic) and 30 benign prostatic hyperplasia patients compared to 50 normal control subjects, using RT‐PCR. Our results of analysis of miR‐21, miR‐141, miR‐18a and miR‐221 in the plasma of PC patients showed that miR‐18a is a powerful discriminator of PC patients from healthy controls as it had the highest AUC (0.966; 95% CI, 0.937–1.000), while miR‐221 provided better differentiation of metastatic from localised PC (sensitivity was 92.9% at 100% specificity), and when we combine miR‐18a and miR‐221 for differentiating patients with MPC, it will increase the sensitivity to 96.4% at a specificity of 100% (AUC, 0.997; 95% CI, 0.988–1.0) (p < .000). This current study recommends that analysis of these miRNAs might have clinical value in enhancing PSA testing.     

他莫昔酚对大鼠前列腺增生动物模型的抑制作用及机理研究

目的 :探讨他莫昔酚抑制前列腺增生的作用机理。　方法 :将Wistar成年大鼠肌肉注射丙酸睾丸酮 ,同时以他莫昔酚常规灌胃。于给药 7、15、30d时分批处死大鼠 ,称取前列腺重量 ,标本做常规病理切片 ,电镜观察前列腺组织细胞结构变化。　结果 :给药 7d时单用睾酮组前列腺指数高于对照组 ,也高于灌胃组。剩余大鼠分 2组 ,其中 1组继续单用他莫昔酚灌胃 ,于第 15、30d时分组处死剩余大鼠。光镜染色观察及电镜扫描均证实他莫昔酚组前列腺上皮细胞及基质的增殖被抑制。　结论 :他莫昔酚通过对雌激素的拮抗作用 ,阻断雌激素在前列腺增生中的作用 ,从而抑制前列腺增生。     

Dickkopf‐related protein 3 promotes pathogenic stromal remodeling in benign prostatic hyperplasia and prostate cancer

BACKGROUND

Compartment‐specific epithelial and stromal expression of the secreted glycoprotein Dickkopf‐related protein (Dkk)‐3 is altered in age‐related proliferative disorders of the human prostate. This study aimed to determine the effect of Dkk‐3 on prostate stromal remodeling that is stromal proliferation, fibroblast‐to‐myofibroblast differentiation and expression of angiogenic factors in vitro.

METHODS

Lentiviral‐delivered overexpression and shRNA‐mediated knockdown of DKK3 were applied to primary human prostatic stromal cells (PrSCs). Cellular proliferation was analyzed by BrdU incorporation ELISA. Expression of Dkk‐3, apoptosis‐related genes, cyclin‐dependent kinase inhibitors and angiogenic factors were analyzed by qPCR, Western blot analysis or ELISA. Fibroblast‐to‐myofibroblast differentiation was monitored by smooth muscle cell actin and insulin‐like growth factor binding protein 3 mRNA and protein levels. The relevance of Wnt/β‐catenin and PI3K/AKT signaling pathways was assessed by cytoplasmic/nuclear β‐catenin levels and phosphorylation of AKT.

RESULTS

Knockdown of DKK3 significantly attenuated PrSC proliferation as well as fibroblast‐to‐myofibroblast differentiation and increased the expression of the vessel stabilizing factor angiopoietin‐1. DKK3 knockdown did not affect subcellular localization or levels of β‐catenin but attenuated AKT phosphorylation in PrSCs. Consistently the PI3K/AKT inhibitor LY294002 mimicked the effects of DKK3 knockdown.

CONCLUSIONS

Dkk‐3 promotes fibroblast proliferation and myofibroblast differentiation and regulates expression of angiopoietin‐1 in prostatic stroma potentially via enhancing PI3K/AKT signaling. Thus, elevated Dkk‐3 in the stroma of the diseased prostate presumably regulates stromal remodeling by enhancing proliferation and differentiation of stromal cells and contributing to the angiogenic switch observed in BPH and PCa. Therefore, Dkk‐3 represents a potential therapeutic target for stromal remodeling in BPH and PCa. Prostate 73: 1441–1452, 2013. © 2013 Wiley‐Liss, Inc. The Prostate published by Wiley Periodicals, Inc.     

New clinical evidence of silodosin,an α1A selective adrenoceptor antagonist,in the treatment for lower urinary tract symptoms

Lower urinary tract symptoms associated with benign prostatic hyperplasia are highly prevalent in older men. Pharmacological treatment is the first‐line treatment for lower urinary tract symptoms associated with benign prostatic hyperplasia. The first choice in the pharmacological treatment for lower urinary tract symptoms associated with benign prostatic hyperplasia is the α₁‐adrenoceptor antagonists. Many α₁‐adrenoceptor antagonists are available in the world. Silodosin is an α₁‐adrenoceptor antagonist developed by Kissei Pharmaceutical, and has a specific selectivity for the α_1A‐adrenoceptor subtype. By antagonizing α_1A‐adrenoceptor in the prostate and urethra, silodosin causes smooth muscle relaxation in the lower urinary tract. As a result of the high affinity for the α_1A‐adrenoceptor than for the α_1B‐adrenoceptor, silodosin minimizes the propensity for blood pressure‐related adverse effects caused by blockade of α_1B‐adrenoceptor. The efficacy and safety of silodosin for treatment of lower urinary tract symptoms associated with benign prostatic hyperplasia was first reported by Japanese investigators in 2006. At present, silodosin is used in many countries. In the present review, we summarize the new clinical evidence for lower urinary tract symptoms associated with benign prostatic hyperplasia and introduce the data supporting the new clinical indications of silodosin.     

结缔组织生长因子在BPH患者膀胱壁中的表达及意义

目的：探讨结缔组织生长因子（CTGF）在BPH患者膀胱壁中的表达,评价CTGF的表达和膀胱出口梗阻所致的膀胱壁纤维化之间的关系。方法：分别留取BPH患者膀胱壁标本25例和对照组膀胱壁标本19例,通过逆转录-聚合酶链反应（RT—PCR）和图像半定量分析,检测组织中CTGF基因表达水平。结果：前列腺增生症和对照组患者膀胱壁中CTGF mRNA指数分别为2.00±0．74、0,79±0．15,两组相比较差异显著,有统计学意义t=8．24（P〈0．01）。结论：CTGF在前列腺增生症患者的膀胱壁组织中表达明显增高,可能是其纤维化的重要促进因素。     

Effects of tadalafil on storage and voiding function in patients with male lower urinary tract symptoms suggestive of benign prostatic hyperplasia: A urodynamic‐based study

Objective

To investigate the effects of tadalafil on storage and voiding function in treatment‐naïve patients with lower urinary tract symptoms suggestive of benign prostatic hyperplasia, based on a urodynamic study.

Methods

This was an open‐labeled, single‐center, prospective study. A total of 80 untreated outpatients with lower urinary tract symptoms suggestive of benign prostatic hyperplasia received tadalafil 5 mg/day for 12 weeks. Subjective symptoms and objective findings on voiding and storage function obtained through urodynamic studies, including cystometry and pressure flow study, were evaluated before and after treatment.

Results

A total of 71 patients with a mean age of 70.2 years and a mean prostate volume of 45.6 mL were included in the analysis. In the International Prostate Symptom Score and Overactive Bladder Symptom Score, mean total scores significantly improved from 18.2 to 13.4 (P < 0.001) and 6.5 to 4.7 (P < 0.001), respectively, after treatment. Mean maximum bladder capacity significantly increased by approximately 35 mL (P < 0.001). Detrusor overactivity disappeared in 15 (39.5%) of 38 patients with detrusor overactivity at baseline (P < 0.001). Mean maximum flow rate on pressure flow study significantly increased from 7.1 to 9.1 mL/s (P < 0.001) and mean bladder outlet obstruction index significantly decreased from 61.3 to 47.1 (P < 0.001).

Conclusions

Treatment with tadalafil 5 mg once daily effectively relieves lower urinary tract symptoms based on objective improvement of storage and voiding function, such as detrusor overactivity and bladder outlet obstruction, in patients with lower urinary tract symptoms suggestive of benign prostatic hyperplasia.     

Demonstration of fibroblast growth factor receptor-I in human prostate by polymerase chain reaction and immunohistochemistry

The expression and localization of fibroblast growth factor receptor-1 were investigated in human prostatic tissues with or without benign hyperplasia. Using a polymerase chain reaction method, we were able to demonstrate that prostatic tissues with benign hyperplasia expressed a significantly higher level of fibroblast growth factor receptor-1 mRNA than normal prostatic tissues (P < 0.01 by Anova). Western blot analysis using an antiserum against the receptor gave 2 bands with molecular weights of about 140 kDa and 80 kDa; these correspond to the expected sizes of the long and secreted forms of the fibroblast growth factor receptor-1, respectively. An immunohistochemical study using the same antiserum further demonstrated that the immunoreactive staining occurred mainly in the basal cells of the glandular epithelium and occasionally in the stromal cells. These results suggest that fibroblast growth factors may influence, at least in part, the proliferation of the epithelial cells seen in benign hyperplasia of human prostate. © 1995 Wiley-Liss, Inc.     

Evaluation and validation of the Core Lower Urinary Tract Symptom Score as an outcome assessment tool for the treatment of benign prostatic hyperplasia: Effects of the α1‐adrenoreceptor antagonist silodosin

We investigated the Core Lower Urinary Tract Symptom Score as an outcome assessment tool for the treatment of lower urinary tract symptoms using silodosin. In addition, the ability of the Core Lower Urinary Tract Symptom Score to detect overactive bladder in male patients with lower urinary tract symptoms was examined. The present study included 241 males with benign prostatic hyperplasia treated at 31 medical facilities between June 2009 and December 2010. All patients were given silodosin, and the effects of silodosin intake were measured using four questionnaires: the Core Lower Urinary Tract Symptom Score, International Prostate Symptom Score, Overactive Bladder Symptom Score and Quality‐of‐Life index. The efficacy of silodosin for treating lower urinary tract symptoms was validated according to the total scores of all four questionnaires weighted equally (P < 0.05). Spearman's ρ among the Core Lower Urinary Tract Symptom Score, International Prostate Symptom Score and Overactive Bladder Symptom Score showed a mild‐high correlation. However, the correlation between the baseline values of the Core Lower Urinary Tract Symptom Score and Quality‐of‐Life index was low in the groups with benign prostatic hyperplasia (ρ = 0.314) and benign prostatic hyperplasia/overactive bladder (ρ = 0.244). Our findings showed the Core Lower Urinary Tract Symptom Score, both its total score and each subscore, is able to show the efficacy of silodosin, similar to other questionnaires. The Core Lower Urinary Tract Symptom Score is also useful for identifying overactive bladder symptoms in patients with benign prostatic hyperplasia. As the Core Lower Urinary Tract Symptom Score does not correlate well with the Quality‐of‐Life index, these two questionnaires might be better used in combination to assess treatment outcomes.     

Upregulation of IL‐17A,CXCL9 and CXCL10 in Early‐Stage Granulomas Induced by Mycobacterium bovis in Cattle

To gain further insight into the immunopathogenesis of bovine tuberculosis (bTB), the cytokine and chemokine expression of cattle experimentally infected with Mycobacterium bovis was analysed in TB granulomas, using immunohistochemistry (IHC) and laser capture microdissection (LCM) followed by qPCR. Immunohistochemistry was conducted for cell types using labelling for CD68, CD3, CD4, CD8, WC1 and CD79a and for the cytokines IFN‐γ, TNF‐α and TGF‐β as well as inducible form of nitric oxide synthase (iNOS). qPCR was conducted for mRNA expression of IFN‐γ, TNF‐α, TGF‐β, IL‐17A, IL‐22, IL‐2, granzyme A and the chemokines CXCL9 and CXCL10. Early stages of granuloma were primarily comprised of epithelioid MΦs expressing high levels of IFN‐γ and iNOS, with significantly upregulated expression of CXCL9 and CXCL10 when compared with control tissue. These chemokines displayed a trend of decreasing mRNA expression as lesion progressed, suggesting a higher level of importance during the early stages of the immune response to mycobacterial infection. IL‐22 levels showed a strong trend of decrease through granuloma development, and IL‐17A was shown to be upregulated, supporting its investigation as a potential biomarker of bTB. The use of LCM and qPCR may prove especially useful for the study of IL‐17A as previous attempts to analyse its expression using IHC and in situ hybridization proved unsuccessful.     

β‐catenin expression and claudin expression pattern as prognostic factors of prostatic cancer progression

OBJECTIVE

To investigate the patterns of expression of the junctional proteins β‐catenin and claudins in different prognostic groups of patients with prostatic cancer, to determine their value as prognostic markers.

PATIENTS AND METHODS

We evaluated the samples of 30 patients who had a radical prostatectomy for organ‐confined cancer (pT2N0M0), men with clinically advanced cancer, and a control group with benign prostatic hyperplasia. Using immunohistochemistry applied to tissue microarrays, each group was evaluated for claudin‐1, ‐2, ‐3, ‐4, ‐5, ‐7, ‐8 and ‐10, and β‐catenin expression.

RESULTS

There were differences among the three groups in the expression of claudin‐1 (P = 0.001), ‐2 (P = 0.014), ‐3 (P = 0.027), ‐4 (P = 0.001), ‐8 (P = 0.001) and β‐catenin (P = 0.002), regardless of Gleason score. By contrast, claudin‐5, ‐7 and ‐10 patterns were not significantly different among the groups. Furthermore, claudin‐1 (P = 0.014) and ‐4 (P = 0.004) could be used to distinguish between those patients who had metastases and those who did not.

CONCLUSIONS

The pattern of claudin expression could be a novel diagnostic marker in re‐classifying adenocarcinomas, and an additional sensitive predictive factor for a clinically poor prognosis. Our results suggest that patients with organ‐confined and advanced cancer are subsets with distinct claudin expression profiles, and that claudin‐4 is related to cellular differentiation in prostate cancer, which is not only the receptor molecule for the Clostridium perfringens enterotoxin, and thus a theoretical future therapeutic target for prostate cancer, but also a marker of progression.