- Email: [email protected]
MP31-18 TESTOSTERONE MODIFIES ALTERATIONS TO DETRUSOR MUSCLE AFTER PARTIAL BLADDER OUTLET OBSTRUCTION IN MICE
THE JOURNAL OF UROLOGYâ
e362
serotonin effects were scrutinized in two different models: Rat ventral prostate explants (VPs) and Human cell line BPH-1. Rat VPs explants (total n¼542) were cultured in vitro (4 days) in different conditions [Control, serotonin, 5-HT1A specific agonist (8-OH-DPAT) and 5-HT1B specific agonist (Anpirtoline)] with and without testosterone supplementation using dose-effect analysis. Total area, external perimeter and branching of explants were determined at D0 and D4. BPH-1 cell line was tested in vitro (3 days) in the same treatment conditions, and cell viability was measured by MTS colorimetric assay. Finally, the androgen receptor was assessed by western-blot analysis. RESULTS: Serotonin receptors are significantly expressed during all rat life-stages. Prostate explants studies revealed that all morphometric measurements were significantly inhibited by serotonin (p<0.001), 8-OH-DPAT (p<0.001) and anpirtoline (p<0.001) both in groups with and without testosterone supplementation, with a dosedependent effect (Fig). Human cell line BPH-1 assays revealed concordant results where serotonin significantly reduced cell viability mostly in groups supplemented with testosterone (Fig). Western blot analysis of VPs explants showed that serotonin significantly decreased AR expression in a dose-dependent way mainly in groups without testosterone supplementation. CONCLUSIONS: Serotonin significantly inhibits non-malignant prostate growth likely by down-regulation of androgen receptors. Thus, we suggest a novel theory for BPH etiology: The neuroendocrine products (ie. serotonin) decreased in prostate transition zone with aging, release the prostate to testosterone-induced constitutive growth resulting in BPH.
Vol. 193, No. 4S, Supplement, Saturday, May 16, 2015
B (AKT), through the quantitative real-time polymerase chain reaction (qRT-PCR) method. RESULTS: After 10 weeks, groups 1 and 2 had less visceral fat when compared to the groups 3 (29.4 vs 37.9 grams; p<0.05) and 4 (31.8 vs 41.9 grams; p<0.05), respectively. According to prostate weight, physical activity decreased prostate growth in both ventral and dorsolateral lobes in normal fed rats (group 1 vs group 3, p<0.05), but this finding was not shown in high fat fed rats (group 2 vs group 4). When comparing normal fed rats (group 1 vs group 3; Graph 1.a), IGF1, IRS1 and AKT gene were more under-expressed in prostate of rats submitted to PA (means 0.22, 0.04 and 0.27 times respectively), and AR was more over-expressed (mean 2.31 times). These patterns of expressions were also shown when we compared high fat fed rats (group 2 vs group 4; Graph 1.b), but the IGF1 underexpression was less pronounced (p<0.001). CONCLUSIONS: PA seems to inhibit prostate IGF1/AKT proliferative axis in both normal and high fat fed rats, and also stimulates AR expression. These mechanisms may play a role in the prevention of BPH proliferative process and prostate inflammation.
Source of Funding: FAPESP - protocol 2014/08368-0 Source of Funding: none
MP31-18 MP31-17
TESTOSTERONE MODIFIES ALTERATIONS TO DETRUSOR MUSCLE AFTER PARTIAL BLADDER OUTLET OBSTRUCTION IN MICE
ROLE OF PHYSICAL ACTIVITY IN THE PREVENTION OF BPH THROUGH INHIBITION OF PROSTATIC IGF1/AKT PROLIFERATIVE AXIS: AN EXPERIMENTAL STUDY IN RATS.
Andrew Flum*, Diana Bowen, Grace Delos Santos, Natalie Kukulka, Paula Firmiss, Robert Dettman, Edward Gong, Chicago, IL
es Fonseca*, Andre M Oliveira, Sabrina T Reis, Ka tia R Fernando Fro ~o Paulo, Leite, William C Nahas, Miguel Srougi, Alberto A Antunes, Sa Brazil
INTRODUCTION AND OBJECTIVES: Bladder outlet obstruction leading to lower urinary tract symptoms is a significant source of morbidity. The bladder has been shown to be hormonally sensitive, but the role of testosterone in the response to bladder injury is unknown. We aim to characterize the effect of testosterone on bladder detrusor muscle following injury from partial bladder outlet obstruction. METHODS: Partial bladder outlet obstruction (pBOO) was surgically created in 8-10 week male CD-1 mice (PO). A subsequent group of male mice was castrated by bilateral orchiectomy at time of pBOO (CPO). Starting at time of surgery testosterone cypionate (10 mg/ g) was administered weekly to another group of castrate pBOO mice (CPOT). Functional evaluation was conducted with voided stain on paper (VSOP) at days 1, 7 and 28. Whole bladders were analyzed at day 28 by bladder weight: body weight ratio and histology. Fibrosis was determined by detrusor smooth muscle:collagen ratio on Masson’s trichrome-stained sections and immunofluorescence staining for COL1a1, vimentin, and FSP-1. RESULTS: PO demonstrated increased bladder weight and increased fibrosis compared to sham (p<0.05 for all comparisons) at
INTRODUCTION AND OBJECTIVES: Sedentarism and obesity have been reported as relevant risk factors for the development of benign prostatic hyperplasia (BPH). In the present work we investigated the role of physical activity in prostatic IGF1/AKT proliferative axis in normal fed and high fat fed rats. METHODS: Twenty eight male Wistar rats with eight weeks old were used in this experiment. They were randomly divided into four groups: 1. physical activity (PA) and normal diet; 2. PA and high fat diet; 3. sedentary (S) and normal diet; 4. S and high fat diet. Food and water were provided ad libitum. Groups 1 and 2 were submitted to a swimming training protocol for 10 weeks, executed five times a week with duration of 60 minutes per day. At the end of the protocol, prostate glands were dissected, weighted and stored. We measured prostatic gene expression levels of insulin-like growth factor 1 (IGF1), IGF1 receptor, insulin receptor substract 1 (IRS1), androgen receptor (AR) and protein kinase
THE JOURNAL OF UROLOGYâ
Vol. 193, No. 4S, Supplement, Saturday, May 16, 2015
day 28 indicating response to bladder outlet obstruction. CPO at day 28 demonstrated improvement in MVV on VSOP compared to PO mice (p<0.05), with lower mean bladder:body weight ratios (p<0.05) and decreased fibrosis by muscle:collagen ratio (p<0.05; figure). Restoration of testosterone in the CPOT group resulted in increased bladder:body weight ratio (p<0.05) with evidence of increased fibrosis demonstrated by lower muscle:collagen ratio (p<0.05; figure) and increased COL1a1 expression on immunofluorescent staining (p<0.05) compared to CPO. CONCLUSIONS: In male mice with partial bladder outlet obstruction castration was associated with larger voided volumes, lower bladder weight and decreased detrusor fibrosis. Replacement of testosterone in castrate mice reversed these effects. This suggests a role for testosterone mediation of the maladaptive changes that occur in detrusor muscle secondary to obstruction. Our findings may provide a novel therapeutic target for the management of lower urinary tract symptoms associated with bladder outlet obstruction.
e363
RESULTS: Hormonal (TþE2) treatment of wild-type mice caused a 30% increase in prostate mass relative to untreated control mice (P < 0.05), which was associated with bladder hypertrophy (P < 0.05). The combination of TþE2 induced similar increased prostate (P < 0.001) and bladder mass in ERbKO mice (P < 0.001) relative to untreated mice. However, in ERaKO mice, TþE2 treatment did not cause increased prostate or bladder mass relative to untreated control ERaKO mice. CONCLUSIONS: Hormonally induced prostate growth was associated with bladder hypertrophy in male mice treated with TþE2. The present results support the conclusion that ERa, but not ERb, is necessary for hormonal induction of prostate growth in male mice treated with TþE2. Source of Funding: F30DK093173
R01DK093690,
T32GMO7356,
MP31-20 THE EFFECT OF SYSTEMIC INFLAMMATION ON PROSTATE VOLUME: A STUDY OF SERUM MAKERS Ryan Werntz*, Wesley Stoller, Brittany Holzhammer, Portland, OR; Jackilen Shannon, portland, OR; Mark Garzotto, Portland, OR
Source of Funding: None
MP31-19 ESTROGEN RECEPTOR-ALPHA IS NECESSARY FOR HORMONAL INDUCTION OF PROSTATE GROWTH IN MALE MICE Tristan Nicholson*, Jalissa Wynder, William Ricke, Madison, WI INTRODUCTION AND OBJECTIVES: Androgens and estrogens act in synergy to promote prostatic growth, and there is an increasing focus on the role of estrogens in benign prostatic hyperplasia (BPH). We have previously shown that adult male mice treated with a combination of testosterone and estradiol (TþE2) develop features of BPH, including bladder hypertrophy, prostate growth and bladder outlet obstruction. Previous work has shown that bladder enlargement in this model depends on intact estrogen receptor-alpha (ERa), but not estrogen receptor-beta (ERb). Therefore, we determined the contribution of these estrogen receptors to prostate growth in male mice treated with TþE2. METHODS: ERa and ERb knockout (KO) male mice were generated on a C57bl/6 background and respective wild-type littermates served as controls. Adult (6-8 weeks) mice underwent surgical implantation of 25 mg T and 2.5 mg E2 slow release hormone pellets for 4 months and were compared to untreated mice. Following euthanasia, urogenital tracts were excised and prostate and bladder mass was determined with a precision balance. Statistical analysis consisted of one-way ANOVA with the Bonferroni post-hoc comparison.
INTRODUCTION AND OBJECTIVES: Diet has a strong effect on systemic inflammation, a process that has been implicated in prostatic carcinogenesis and other cancer types. However, it is unknown whether this mechanism affects benign prostatic hyperplasia e an entity in which inflammation has also been implicated to play a causative role. The goal of this study was to evaluate the effects of a pro-inflammatory diet and serum markers of systemic inflammation on prostate volume. METHODS: We conducted an analysis of data from the Diet and Prostate Cancer Risk case-control study conducted from 2001 to 2006 at the Portland Veterans Affairs Medical Center. We studied the effect of dietary-induced inflammation on prostate volume from 230 biopsy-negative patients. Dietary inflammation was measured by the dietary inflammatory index (DII) score; a measure of dietary inflammatory potential. Systemic markers studied included plasma IL-6 and CRP. Multivariable logistic regression analysis was used to measure the effect of systemic inflammation (DII, IL-6, CRP) and clinical factors (age, BMI and PSA) on prostate volume. RESULTS: Mean prostate volume was 52.6 25.5 cc, mean PSA was 6.4 4.6 ng/ml and BMI was29.3 5.0 kg/m2. Mean DII score -0.81 2.1, mean IL-6 was 2.62.9 pg/ml and mean CRP was 2.9 4.8 We found no significant association between DII score (p ¼.283), IL-6 (.891) or CRP (.154) with prostate volume. In contrast, significant predictors of volume included age (p ¼.000), PSA (p ¼.015), and BMI (p ¼.001). The overall logistic regression model improved with removal of the DII and IL-6 (p¼.238 to.848, Hosmer “goodness-offit” test) CONCLUSIONS: DII score and plasma markers of systemic inflammation were not associated with prostate volume in this study. However Age, PSA and BMI were strong predictors of prostate volume. These data indicate that while local inflammation may play a role in the genesis of benign prostatic tissue, these data do not support a role for systemic inflammation. Source of Funding: National Institute of Health
e362
serotonin effects were scrutinized in two different models: Rat ventral prostate explants (VPs) and Human cell line BPH-1. Rat VPs explants (total n¼542) were cultured in vitro (4 days) in different conditions [Control, serotonin, 5-HT1A specific agonist (8-OH-DPAT) and 5-HT1B specific agonist (Anpirtoline)] with and without testosterone supplementation using dose-effect analysis. Total area, external perimeter and branching of explants were determined at D0 and D4. BPH-1 cell line was tested in vitro (3 days) in the same treatment conditions, and cell viability was measured by MTS colorimetric assay. Finally, the androgen receptor was assessed by western-blot analysis. RESULTS: Serotonin receptors are significantly expressed during all rat life-stages. Prostate explants studies revealed that all morphometric measurements were significantly inhibited by serotonin (p<0.001), 8-OH-DPAT (p<0.001) and anpirtoline (p<0.001) both in groups with and without testosterone supplementation, with a dosedependent effect (Fig). Human cell line BPH-1 assays revealed concordant results where serotonin significantly reduced cell viability mostly in groups supplemented with testosterone (Fig). Western blot analysis of VPs explants showed that serotonin significantly decreased AR expression in a dose-dependent way mainly in groups without testosterone supplementation. CONCLUSIONS: Serotonin significantly inhibits non-malignant prostate growth likely by down-regulation of androgen receptors. Thus, we suggest a novel theory for BPH etiology: The neuroendocrine products (ie. serotonin) decreased in prostate transition zone with aging, release the prostate to testosterone-induced constitutive growth resulting in BPH.
Vol. 193, No. 4S, Supplement, Saturday, May 16, 2015
B (AKT), through the quantitative real-time polymerase chain reaction (qRT-PCR) method. RESULTS: After 10 weeks, groups 1 and 2 had less visceral fat when compared to the groups 3 (29.4 vs 37.9 grams; p<0.05) and 4 (31.8 vs 41.9 grams; p<0.05), respectively. According to prostate weight, physical activity decreased prostate growth in both ventral and dorsolateral lobes in normal fed rats (group 1 vs group 3, p<0.05), but this finding was not shown in high fat fed rats (group 2 vs group 4). When comparing normal fed rats (group 1 vs group 3; Graph 1.a), IGF1, IRS1 and AKT gene were more under-expressed in prostate of rats submitted to PA (means 0.22, 0.04 and 0.27 times respectively), and AR was more over-expressed (mean 2.31 times). These patterns of expressions were also shown when we compared high fat fed rats (group 2 vs group 4; Graph 1.b), but the IGF1 underexpression was less pronounced (p<0.001). CONCLUSIONS: PA seems to inhibit prostate IGF1/AKT proliferative axis in both normal and high fat fed rats, and also stimulates AR expression. These mechanisms may play a role in the prevention of BPH proliferative process and prostate inflammation.
Source of Funding: FAPESP - protocol 2014/08368-0 Source of Funding: none
MP31-18 MP31-17
TESTOSTERONE MODIFIES ALTERATIONS TO DETRUSOR MUSCLE AFTER PARTIAL BLADDER OUTLET OBSTRUCTION IN MICE
ROLE OF PHYSICAL ACTIVITY IN THE PREVENTION OF BPH THROUGH INHIBITION OF PROSTATIC IGF1/AKT PROLIFERATIVE AXIS: AN EXPERIMENTAL STUDY IN RATS.
Andrew Flum*, Diana Bowen, Grace Delos Santos, Natalie Kukulka, Paula Firmiss, Robert Dettman, Edward Gong, Chicago, IL
es Fonseca*, Andre M Oliveira, Sabrina T Reis, Ka tia R Fernando Fro ~o Paulo, Leite, William C Nahas, Miguel Srougi, Alberto A Antunes, Sa Brazil
INTRODUCTION AND OBJECTIVES: Bladder outlet obstruction leading to lower urinary tract symptoms is a significant source of morbidity. The bladder has been shown to be hormonally sensitive, but the role of testosterone in the response to bladder injury is unknown. We aim to characterize the effect of testosterone on bladder detrusor muscle following injury from partial bladder outlet obstruction. METHODS: Partial bladder outlet obstruction (pBOO) was surgically created in 8-10 week male CD-1 mice (PO). A subsequent group of male mice was castrated by bilateral orchiectomy at time of pBOO (CPO). Starting at time of surgery testosterone cypionate (10 mg/ g) was administered weekly to another group of castrate pBOO mice (CPOT). Functional evaluation was conducted with voided stain on paper (VSOP) at days 1, 7 and 28. Whole bladders were analyzed at day 28 by bladder weight: body weight ratio and histology. Fibrosis was determined by detrusor smooth muscle:collagen ratio on Masson’s trichrome-stained sections and immunofluorescence staining for COL1a1, vimentin, and FSP-1. RESULTS: PO demonstrated increased bladder weight and increased fibrosis compared to sham (p<0.05 for all comparisons) at
INTRODUCTION AND OBJECTIVES: Sedentarism and obesity have been reported as relevant risk factors for the development of benign prostatic hyperplasia (BPH). In the present work we investigated the role of physical activity in prostatic IGF1/AKT proliferative axis in normal fed and high fat fed rats. METHODS: Twenty eight male Wistar rats with eight weeks old were used in this experiment. They were randomly divided into four groups: 1. physical activity (PA) and normal diet; 2. PA and high fat diet; 3. sedentary (S) and normal diet; 4. S and high fat diet. Food and water were provided ad libitum. Groups 1 and 2 were submitted to a swimming training protocol for 10 weeks, executed five times a week with duration of 60 minutes per day. At the end of the protocol, prostate glands were dissected, weighted and stored. We measured prostatic gene expression levels of insulin-like growth factor 1 (IGF1), IGF1 receptor, insulin receptor substract 1 (IRS1), androgen receptor (AR) and protein kinase
THE JOURNAL OF UROLOGYâ
Vol. 193, No. 4S, Supplement, Saturday, May 16, 2015
day 28 indicating response to bladder outlet obstruction. CPO at day 28 demonstrated improvement in MVV on VSOP compared to PO mice (p<0.05), with lower mean bladder:body weight ratios (p<0.05) and decreased fibrosis by muscle:collagen ratio (p<0.05; figure). Restoration of testosterone in the CPOT group resulted in increased bladder:body weight ratio (p<0.05) with evidence of increased fibrosis demonstrated by lower muscle:collagen ratio (p<0.05; figure) and increased COL1a1 expression on immunofluorescent staining (p<0.05) compared to CPO. CONCLUSIONS: In male mice with partial bladder outlet obstruction castration was associated with larger voided volumes, lower bladder weight and decreased detrusor fibrosis. Replacement of testosterone in castrate mice reversed these effects. This suggests a role for testosterone mediation of the maladaptive changes that occur in detrusor muscle secondary to obstruction. Our findings may provide a novel therapeutic target for the management of lower urinary tract symptoms associated with bladder outlet obstruction.
e363
RESULTS: Hormonal (TþE2) treatment of wild-type mice caused a 30% increase in prostate mass relative to untreated control mice (P < 0.05), which was associated with bladder hypertrophy (P < 0.05). The combination of TþE2 induced similar increased prostate (P < 0.001) and bladder mass in ERbKO mice (P < 0.001) relative to untreated mice. However, in ERaKO mice, TþE2 treatment did not cause increased prostate or bladder mass relative to untreated control ERaKO mice. CONCLUSIONS: Hormonally induced prostate growth was associated with bladder hypertrophy in male mice treated with TþE2. The present results support the conclusion that ERa, but not ERb, is necessary for hormonal induction of prostate growth in male mice treated with TþE2. Source of Funding: F30DK093173
R01DK093690,
T32GMO7356,
MP31-20 THE EFFECT OF SYSTEMIC INFLAMMATION ON PROSTATE VOLUME: A STUDY OF SERUM MAKERS Ryan Werntz*, Wesley Stoller, Brittany Holzhammer, Portland, OR; Jackilen Shannon, portland, OR; Mark Garzotto, Portland, OR
Source of Funding: None
MP31-19 ESTROGEN RECEPTOR-ALPHA IS NECESSARY FOR HORMONAL INDUCTION OF PROSTATE GROWTH IN MALE MICE Tristan Nicholson*, Jalissa Wynder, William Ricke, Madison, WI INTRODUCTION AND OBJECTIVES: Androgens and estrogens act in synergy to promote prostatic growth, and there is an increasing focus on the role of estrogens in benign prostatic hyperplasia (BPH). We have previously shown that adult male mice treated with a combination of testosterone and estradiol (TþE2) develop features of BPH, including bladder hypertrophy, prostate growth and bladder outlet obstruction. Previous work has shown that bladder enlargement in this model depends on intact estrogen receptor-alpha (ERa), but not estrogen receptor-beta (ERb). Therefore, we determined the contribution of these estrogen receptors to prostate growth in male mice treated with TþE2. METHODS: ERa and ERb knockout (KO) male mice were generated on a C57bl/6 background and respective wild-type littermates served as controls. Adult (6-8 weeks) mice underwent surgical implantation of 25 mg T and 2.5 mg E2 slow release hormone pellets for 4 months and were compared to untreated mice. Following euthanasia, urogenital tracts were excised and prostate and bladder mass was determined with a precision balance. Statistical analysis consisted of one-way ANOVA with the Bonferroni post-hoc comparison.
INTRODUCTION AND OBJECTIVES: Diet has a strong effect on systemic inflammation, a process that has been implicated in prostatic carcinogenesis and other cancer types. However, it is unknown whether this mechanism affects benign prostatic hyperplasia e an entity in which inflammation has also been implicated to play a causative role. The goal of this study was to evaluate the effects of a pro-inflammatory diet and serum markers of systemic inflammation on prostate volume. METHODS: We conducted an analysis of data from the Diet and Prostate Cancer Risk case-control study conducted from 2001 to 2006 at the Portland Veterans Affairs Medical Center. We studied the effect of dietary-induced inflammation on prostate volume from 230 biopsy-negative patients. Dietary inflammation was measured by the dietary inflammatory index (DII) score; a measure of dietary inflammatory potential. Systemic markers studied included plasma IL-6 and CRP. Multivariable logistic regression analysis was used to measure the effect of systemic inflammation (DII, IL-6, CRP) and clinical factors (age, BMI and PSA) on prostate volume. RESULTS: Mean prostate volume was 52.6 25.5 cc, mean PSA was 6.4 4.6 ng/ml and BMI was29.3 5.0 kg/m2. Mean DII score -0.81 2.1, mean IL-6 was 2.62.9 pg/ml and mean CRP was 2.9 4.8 We found no significant association between DII score (p ¼.283), IL-6 (.891) or CRP (.154) with prostate volume. In contrast, significant predictors of volume included age (p ¼.000), PSA (p ¼.015), and BMI (p ¼.001). The overall logistic regression model improved with removal of the DII and IL-6 (p¼.238 to.848, Hosmer “goodness-offit” test) CONCLUSIONS: DII score and plasma markers of systemic inflammation were not associated with prostate volume in this study. However Age, PSA and BMI were strong predictors of prostate volume. These data indicate that while local inflammation may play a role in the genesis of benign prostatic tissue, these data do not support a role for systemic inflammation. Source of Funding: National Institute of Health