Ventral Prostate Lobes Research Articles

The morphological structure of prostate gland under the condition of experimental prostatopathy and after using of cholecalciferol in the different schemes of hypofertility correction

The impact of negative factors, stress, and modern living conditions damages men's health and leads to infertility. Prostatitis is often a cause of hypofertility. It is now shown that vitamin D may play a role in regulating the functioning of reproductive system organs. The aim of the study was to determine the effect of cholecalciferol on the histological structure of the prostate gland in rats with experimental prostatitis and after its application alone or in combination with a prostate protector. Experimental prostatitis was induced by cold intraoperative damage to the prostate gland. To correct prostatitis, vitamin D3 (cholecalciferol) was administered orally at a dose of 4000 IU. The prostate protector (Prostatilen, Pr) and its pharmaceutical composition, as well as vitamin D3, were administered rectally. Rats with modeled prostatitis were divided into groups: EP (cold experimental prostatitis without treatment); EP + seed oil (on the background of experimental prostatitis, animals received a solvent – apricot kernel oil); EP + vit D3 (per os) (on the background of experimental prostatitis, animals received vitamin D3); EP + Pr (rec) (on the background of experimental prostatitis, males were administered Prostatilen per rectum); EP + vit. D3 (per os) + Pr (rec) (on the background of experimental prostatitis, animals received vitamin D3 (per os) and Prostatilen gel (per rectum)); EP + (vit. D3 + Pr) (rec) (on the background of experimental prostatitis, rats were administered Prostatilen gel with vitamin D3 per rectum). Intact animals (Intact group) and sham-operated rats (Control group) were used as controls. Paraffin sections of the prostate gland were stained with hematoxylin, eosin, and Van Gieson's method. In addition to the review microscopy of the ventral lobes of the prostate gland and the isthmus between them, the power of histochemical reactions was measured, the severity of inflammation and fibrosis was assessed, the number of terminal sections of the prostatic glands with a visually unchanged state, with lumen expansion, and with wall destruction was counted, the longitudinal diameter of the acini and the height of the epithelial cells of the prostatic glands were measured. Statistical analysis of the results was performed using the standard software package "Statistica 6.0" with the use of Student's t-test and nonparametric analog of one-way analysis of variance – Kruskal-Wallis test, as well as Mann-Whitney test. It was found that rats with experimental prostatitis exhibit pronounced changes in the morphological structure of the prostate gland. The prostate-protective effect of vitamin D per os at a dose of 4000 IU was established, which reduced the manifestations of atrophic and destructive processes, signs of tissue inflammation, and coarsening of the prostate gland stroma. Signs of fibrosis development in the prostate gland in males of this group were reduced, and the number of destructive changes and the longitudinal diameter of the terminal sections of the prostatic glands of prostate ventral lobe in rats with experimental prostatitis were decreased. Thus, the addition of cholecalciferol to the basic therapy for infertility has a more pronounced corrective effect on the morphological structure of the prostate than the separate use of cholecalciferol and the prostate protector. Combining basic therapy with vitamin D enhances the prostate-protective properties of the latter and is promising for restoring reproductive function overall.

P. gingivalis in oral-prostate axis exacerbates benign prostatic hyperplasia via IL-6/IL-6R pathway.

Benign prostatic hyperplasia (BPH) is the most common disease in elderly men. There is increasing evidence that periodontitis increases the risk of BPH, but the specific mechanism remains unclear. This study aimed to explore the role and mechanism of the key periodontal pathogen Porphyromonas gingivalis (P. gingivalis) in the development of BPH. The subgingival plaque (Sp) and prostatic fluid (Pf) of patients with BPH concurrent periodontitis were extracted and cultured for 16S rDNA sequencing. Ligature-induced periodontitis, testosterone-induced BPH and the composite models in rats were established. The P. gingivalis and its toxic factor P. gingivalis lipopolysaccharide (P.g-LPS) were injected into the ventral lobe of prostate in rats to simulate its colonization of prostate. P.g-LPS was used to construct the prostate cell infection model for mechanism exploration. P. gingivalis, Streptococcus oralis, Capnocytophaga ochracea and other oral pathogens were simultaneously detected in the Pf and Sp of patients with BPH concurrent periodontitis, and the average relative abundance of P. gingivalis was found to be the highest. P. gingivalis was detected in both Pf and Sp in 62.5% of patients. Simultaneous periodontitis and BPH synergistically aggravated prostate histological changes. P. gingivalis and P.g-LPS infection could induce obvious hyperplasia of the prostate epithelium and stroma (epithelial thickness was 2.97- and 3.08-fold that of control group, respectively), and increase of collagen fibrosis (3.81- and 5.02-fold that of control group, respectively). P. gingivalis infection promoted prostate cell proliferation, inhibited apoptosis, and upregulated the expression of inflammatory cytokines interleukin-6 (IL-6; 4.47-fold), interleukin-6 receptor-α (IL-6Rα; 5.74-fold) and glycoprotein 130 (gp130; 4.47-fold) in prostatic tissue. P.g-LPS could significantly inhibit cell apoptosis, promote mitosis and proliferation of cells. P.g-LPS activates the Akt pathway through IL-6/IL-6Rα/gp130 complex, which destroys the imbalance between proliferation and apoptosis of prostate cells, induces BPH. P. gingivalis was abundant in the Pf of patients with BPH concurrent periodontitis. P. gingivalis infection can promote BPH, which may affect the progression of BPH via inflammation and the Akt signaling pathway.

Histological study on the toxic effect of titanium dioxide nanoparticles on the prostate gland of adult albino rats and the protective role of coenzyme Q10

Background Titanium dioxide nanoparticles have numerous clinical and commercial applications. Coenzyme Q10 has antioxidant and free radical-scavenging capabilities in addition to functions like vitamins. Aim To identify changes in the ventral prostatic lobe of adult albino rats exposed to titanium dioxide nanoparticles and to evaluate the possible protective role of coenzyme Q 10. Materials and methods Forty adult male albino rats were used in this study. Group I was the control, Group II received coenzyme Q10 at a dose of 20 mg/kg, Group III received titanium dioxide nanoparticles at a dose of 600 mg/kg, and Group IV received both coenzyme Q10 and nanoparticles. Prostate glands sections were stained with hematoxylin and eosin. A morphometric study and statistical analysis were performed on the epithelial height. Results Sections of group III showed some histological abnormalities in the form of widely spaced prostatic acini, an increase in the amount of connective tissue stroma, with numerous dilated blood vessels and perivascular cellular infiltration. The mean height of the prostatic epithelium in group III was significantly higher than in the control group. Prostatic epithelial height was found to be significantly lower in group IV as compared to group III. Conclusion Titanium dioxide nanoparticles induce changes in the prostatic ventral lobe which can be ameliorated by coenzyme Q10 supplementation.

Broccoli sprouts delay prostate cancer formation and decrease prostate cancer severity with a concurrent decrease in HDAC3 protein expression in TRAMP mice

Background: Cruciferous vegetables have been associated with the chemoprevention of cancer. Epigenetic regulators have been identified as important targets for prostate cancer chemoprevention. Treatment of human prostate cancer cells with sulforaphane (SFN), a chemical from broccoli and broccoli sprouts, inhibits epigenetic regulators such as histone deacetylase (HDAC) enzymes, but it is not known whether consumption of a diet high in broccoli sprouts impacts epigenetic mechanisms in an in vivo model of prostate cancer. Objective: In the transgenic adenocarcinoma of the mouse prostate (TRAMP) model, we tested the hypothesis that a broccoli sprout diet suppresses prostate cancer, inhibits HDAC expression, alters histone modifications, and changes the expression of genes regulated by HDACs. Methods: TRAMP mice were fed a 15% broccoli sprout, or control AIN93G diet; tissue samples were collected at 12 and 28 weeks of age. Results: Mice fed broccoli sprouts had detectable levels of SFN metabolites in liver, kidney, colon, and prostate tissues. Broccoli sprouts reduced prostate cancer incidence, and progression to invasive cancer by 11- and 2.4-fold at 12 and 28 weeks of age, respectively. There was a significant decline in HDAC3 protein expression in the epithelial cells of prostate ventral and anterior lobes at 12 weeks. Broccoli sprout consumption also decreased histone H3 lysine 9 tri-methylation in the ventral lobe (12 week), and decreased histone H3 lysine 18 acetylation in all prostate lobes (28 weeks). A decline in p16 mRNA levels, a gene regulated by HDAC3, was associated with broccoli sprouts consumption, but no significant changes were noted at the protein level. Conclusions: Broccoli sprout intake caused a decline in prostate cancer occurrence and HDAC3 protein expression in the prostate, extending prior work that implicated loss of HDAC3/corepressor interactions as a key preventive mechanism by SFN in vivo.

Aqueous extract of Teucrium polium ameliorates diabetes and induced-prostatic complication.

Prostatic complications are common among diabetic patients. Previous research demonstrates that Teucrium polium (T. polium) has beneficial effects in diabetic cases. This study, therefore, aimed to evaluate the impacts of T. polium aqueous extract on the prostate of diabetic rats. Diabetes was induced in male Wistar rats by intraperitoneal injection of streptozotocin (50mg/kg). a total of 40 Rats were randomly divided into the following groups: Control, Control + TP100 (TP100), Diabetic, Diabetic + TP100 (DTP100) and Diabetic + TP200 (DTP200). The intervention was done orally once per day for 56days (8weeks). An oral glucose tolerance test was conducted, glucose and insulin levels were assessed. Microscopic features of the ventral prostatic lobe were evaluated pathologically. T. polium at both doses significantly reduced glucose levels in an insulin-independent pathway. T. polium at both doses significantly improved prostate weight, prostate epithelium height, and prostate secretory activity in comparison with the diabetic group. Interestingly, treatment of T. polium to healthy rats led to decreased epithelial height. It could be deduced that T. polium has useful impacts on glucose control and may prevent prostatic complications. The online version contains supplementary material available at 10.1007/s40200-022-00979-4.

Comparison of the Effects of Allium cepa L. Extract Together with Insulin on Sperm Parameters in Diabetic Rats

Background: In diabetic patients, uncontrolled blood sugar causes disorders in various systems of the body in the long term. The reproductive system is one of these susceptible systems. It is known that diabetes can adversely affect spermatogenesis. The use of medicinal plants in the treatment of various diseases has been discussed by many researchers for a long time. Onion, scientifically known as Allium cepa L., contains antioxidants. Insulin is also a drug used to control blood sugar in diabetic patients. Objectives: This study aimed to evaluate and compare in vivo antidiabetic activities of hydroalcoholic onion seed extract together with insulin in diabetic rats. Methods: In this study, diabetes was induced in rats with streptozotocin (60 mg/kg). Fifty animals were equally divided into five groups: nondiabetic control (group 1); diabetic control (group 2); diabetic rats receiving streptozotocin plus insulin (group 3); and diabetic rats treated with 200 and 400 mg/kg of Allium cepa L. seed extract by gavage for four weeks (groups 4 and 5). At the end of the study, the prostate ventral lobe was removed and processed for histological studies. Next, sperm parameters from the tail of the left epididymis, biochemical parameters, and histopathological changes were analyzed and compared. Results: The sperm parameters of diabetic rats receiving 200 and 400 mg/kg of Allium cepa L. extract showed a significant increase compared to the diabetic control group. Conclusions: Administration of Allium cepa L. extract as a strong antioxidant was adequate to compensate for the toxic effects of streptozotocin and increase the motility of sperms.

Protective effect of the association of curcumin with piperine on prostatic lesions: New perspectives on BPA-induced carcinogenesis

Bisphenol A (BPA) is a chemical agent which can exert detrimental effects on the male reproductive system, especially the prostate gland. In this study we described the efficacy of the dietary agent curcumin, alone or combined with piperine, to suppress the impact of BPA on the prostate. Adult gerbils were divided into nine experimental groups (n = 7 each group), regarding control (water and oil), exposed to BPA (50 μg/kg/day in water) or curcumin (100 mg/kg) and/or piperine (20 mg/kg). To evaluate the effects of the phytotherapic agents, the other groups received oral doses every two days, BPA plus curcumin (BCm), piperine (BP), and curcumin + piperine (BCmP). BPA promoted prostatic inflammation and morphological lesions in ventral and dorsolateral prostate lobes, associated with an increase in androgen receptor-positive cells and nuclear atypia, mainly in the ventral lobe. Curcumin and piperine helped to minimize these effects. BPA plus piperine or curcumin showed a reduction in nuclear atypical phenotype, indicating a beneficial effect of phytochemicals. Thus, these phytochemicals minimize the deleterious action of BPA in prostatic lobes, especially when administered in association. The protective action of curcumin and piperine consumption is associated with weight loss, anti-inflammatory potential, and control of prostate epithelial cell homeostasis.

Screening of anti-chronic nonbacterial prostatitis activity of different extractions of the aerial part of Glycyrrhiza uralensis, and network pharmacology research.

In the present study, anti-chronic nonbacterial prostatitis (CNP) pharmacological experiments using water and ethanol extraction of the aerial parts of Glycyrrhiza uralensis were performed to select the best active parts by comparing their efficacy in a CNP model established by injecting carrageenin into the ventral lobe of rat prostate. The anti-CNP activities and expression of serum inflammatory factors in rats were also analyzed. A Protein-Protein Interaction network was constructed, and core targets were screened using topology and analyzed using Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. Water and ethanol extraction exhibited good inhibitory effect on the pathological changes of the prostate tissue, the expression of inflammatory factors and fibrosis factors in CNP rats, whereas no differences were observed compared with the positive control drug. Water extraction was more effective and significantly reduced PGE2 expression (P<0.05). Network pharmacology assays showed 15 active components in the aerial part of Glycyrrhiza uralensis, and 9 key CNP therapeutic targets of the aerial parts of Glycyrrhiza uralensis were identified. The effect of water exraction on chronic prostatitis rats was significant. The aerial part of Glycyrrhiza uralensis downregulated the levels of inflammatory factors and inhibited proinflammatory gene transcription, reduced oxidative stress response, inhibited cell survival pathways, regulated sex hormone levels, prevented immunostimulation and attenuated inflammation. This study provides a theoretical reference for the development of anti-CNP agents, and offers a novel methodology for identifying and clarifying the mechanisms underlying the efficacy of the anti-CNP components in the aerial part of Glycyrrhiza uralensis.

PD11-04 LOSS OF OSTEOPONTIN LEADS TO THE RESOLUTION OF E. COLI-INDUCED PROSTATIC INFLAMMATION AND FIBROSIS

You have accessJournal of UrologyBenign Prostatic Hyperplasia: Basic Research & Pathophysiology (PD11)1 Sep 2021PD11-04 LOSS OF OSTEOPONTIN LEADS TO THE RESOLUTION OF E. COLI-INDUCED PROSTATIC INFLAMMATION AND FIBROSIS Petra Popovics, Asha Jain, Francesca Van, Hannah Ruetten, Mark Cadena, Kristen S. Uchtmann, Chad M. Vezina, and William A. Ricke Petra PopovicsPetra Popovics More articles by this author , Asha JainAsha Jain More articles by this author , Francesca VanFrancesca Van More articles by this author , Hannah RuettenHannah Ruetten More articles by this author , Mark CadenaMark Cadena More articles by this author , Kristen S. UchtmannKristen S. Uchtmann More articles by this author , Chad M. VezinaChad M. Vezina More articles by this author , and William A. RickeWilliam A. Ricke More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000001986.04AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Fibrogenic and inflammatory processes in the prostate are linked to the development of lower urinary tract symptoms (LUTS) in men, but the molecular mechanism is unknown. We previously established that osteopontin (OPN), a pro-fibrotic cytokine, is more abundant in the prostate of men with LUTS compared to prostate tissue with normal histology. We also identified that OPN secretion is stimulated by inflammatory cytokines in prostate stromal cells and hypothesized that OPN exacerbates the prostatic inflammatory environment. The current study investigates whether the lack of OPN ameliorates inflammation-induced prostatic fibrosis and urinary dysfunction in mice following transurethral instillation of E. coli. METHODS: Uropathogenic Escherichia coli (UTI89) or saline (control) was instilled via a transurethral catheter (OD 0.8, 100 ul) two times, 3 days apart to C57BL/6J (WT) or B6.129S6(Cg)-Spp1tm1Blh/J (OPN-KO) mice. Urinary function was assessed by weekly void spot assays (VSA). Prostatic collagen was visualized with picrosirius red (PSR) staining and inflammation by immunohistochemistry of the pan-leukocyte marker CD45. RNA-Seq analysis was performed from the ventral prostate lobe. RESULTS: Urinary frequency significantly increased (2.05-fold, p=0.0097) 33 days after E. coli instillation in WT but not in OPN-KO mice. One week after infection, the ventral and dorsal prostate lobes were significantly inflamed (more CD45 cells) and collagen fibers were thicker in both WT and OPN-KO mice. In contrast, 2 months after instillation, OPN-KO mouse prostates contained significantly less total collagen and inflammation compared to WT mice. RNAseq analysis identified that E. coli-instilled WT mice expressed more inflammatory and fibrotic marker RNAs, such as Col3a1, Il1b, MMP3, Mmp7 and Cxcl12 than OPN-KO mice. CONCLUSIONS: Our results show that by preventing the inflammation-related increase in OPN levels, mice undergo accelerated recovery from fibrosis and do not develop chronic inflammation. This suggests that drugs targeting OPN signaling could be beneficial for LUTS patients whose symptoms are primarily linked to fibrosis. Source of Funding: The study was supported by an NIH NIDDK K01 (K01DK127150) and a K12 award (K12DK100022-06, both to PP) and by U54 DK104310 (to WAR and CMV) © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e198-e199 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Petra Popovics More articles by this author Asha Jain More articles by this author Francesca Van More articles by this author Hannah Ruetten More articles by this author Mark Cadena More articles by this author Kristen S. Uchtmann More articles by this author Chad M. Vezina More articles by this author William A. Ricke More articles by this author Expand All Advertisement Loading ...

Abstract 2497: Inositol Polyphosphate 4-Phosphatase Type-II B protects mice from benign prostatic hyperplasia

Abstract BPH is the most common tumor-like condition that afflicts more than 50% of men between the ages of 51-60 and over 90% in men older than 80. While connection between obesity and BPH is well established, the driving signaling pathways are not clear. BPH is a nonmalignant enlargement of prostate epithelium that requires androgen receptor (AR) signaling. INPP4B and EZH2 are highly expressed in prostate epithelium where they regulate AR signaling and play an important role in prostate cancer etiology. EZH2 is an epigenetic silencer that methylates histone H3 on K9 and K27 residues and INPP4B is a dual specificity phosphatase that dephosphorylates PI(3,4)P2 and PI(4,5)P2, membrane lipids that activate Akt and PKC signaling pathways respectively. We discovered that Inpp4b-/- male mice develop BPH at 3 months old of age and subsequently prostate intraepithelial neoplasia (PIN) when fed high fat diet. While expression of AR in the prostates of these mice is similar, AR activity is reduced. The loss of INPP4B caused reduction in EZH2 protein levels in prostate cancer cell lines and in mouse prostate. Inhibition of Akt and PKC signaling reduced EZH2 levels in ventral and dorsolateral prostate lobes of Inpp4b-/- males, suggesting that INPP4B upregulates EZH2 levels by suppressing Akt and PKC signaling pathways. Consistent with decline in EZH2 levels, Inpp4b-/- males have reduced levels of H3K27me3 and H3K9me2 in the ventral prostate, and high fat diet exacerbated this phenotype. Our data suggests that INPP4B modulates epigenetic profile of prostate epithelium by stabilizing EZH2 levels. Both loss of INPP4B and high fat diet treatment lead to decline in EZH2 levels and activity and contribute to development of BPH in mouse model. Citation Format: Yasemin Ceyhan, Manqi Zhang, Irina U. Agoulnik. Inositol Polyphosphate 4-Phosphatase Type-II B protects mice from benign prostatic hyperplasia [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2497.

Effect of Moderate and High Intensity Exercise on Hypoxia and Radiosensitivity in Tumor‐Bearing Rats

Background Prostate tumors typically have hypoxic regions that are resistant to traditional radiation therapy. Prior evidence from our laboratory has demonstrated improved perfusion and reduced hypoxia of prostate tumors with moderate intensity exercise training; such changes have potential to enhance radiation therapy. However, there is a dearth of evidence for the magnitude of alterations of intra-tumor hypoxia markers with exercise, as well as the ideal intensity of exercise for combating hypoxia in solid tumors. Using a pre-clinical orthotopic model of prostate cancer, we tested the hypothesis that exercise training at both moderate and high intensities would result in a decrease in hypoxia assessed via immunohistochemical analysis of hypoxic markers. Further, that enhanced radiotherapy responses would occur in exercise trained tumor bearing animals compared to sedentary counterparts. Methods Immunocompetent male Copenhagen rats aged 6 months (n=48) were injected orthotopically (ventral lobe of prostate) with 1x105 Dunning R-3327 AT-1 prostate adenocarcinoma cells. All animals were first acclimated to treadmill exercise and randomized into three groups: tumor bearing sedentary (TBS, n=20), moderate intensity training, (TBMIT, n=12), or high intensity training (TBHIT, n=16). After 7 days of recovery from surgery, both exercise groups began progressive exercise training on a motorized treadmill at either 20/min with a 5% incline, or 30m/min 15% incline for 10 minutes a day progressing to 60 min/day for TBMIT and TBHIT, respectively, for approximately 5 weeks. Immunohistochemical analysis and whole-body ionizing radiation (2 Gy) were performed at the end of the 5-week period of training followed by clonogenic cell survival assay to assess survival fraction. Results There were no significant differences (p>0.05) in tumor mass between groups (TBS 7.8±1.1; TBMIT 7.2±1.3; TBHIT 6.4±0.9 g). Overall expression levels of hypoxia inducible factor 1-alpha (HIF-1a, n=40) were significantly greater in the TBS group vs both the TBMIT and TBHIT (TBS 37.9±7.4; TBMIT 25.5±6.1; TBHIT 14.4±5.7%, p<0.05), with a trend for increased expression in TBMIT vs TBHIT (p=0.059). The survival fraction (n=8) was also significantly higher in TBS when compared to both TBMIT and TBHIT (TBS 39.9±5.3; TBMIT 23.2±3.2; TBHIT 20.7±4.9%, p<0.05). Conclusion This study suggests that compared to sedentary counterparts, both chronic moderate and high intensity exercise training modify the tumor microenvironment (decreased levels of HIF-1a) favorably. Given exercise therapy is becoming increasingly recommended to mitigate fatigue and increase quality of life in patients, a potential therapeutic component could add to the efficacy of exercise prescription in a clinical setting.

Pesticide Dichlorvos Promotes Morphological Alternations in the Rat Prostate

The complex process of carcinogenesis can be modulated by the action of environmental endocrine disruptors, such as the organophosphate pesticide Dichlorvos (DDVP), capable of interfering in homeostasis of different organs and leading to the development of neoplastic lesions. Considering that prostate cancer is the second most common neoplasm in men, worldwide, it is emphasized the importance of studying morphological alterations using animal models. In rodents, tumor initiation can be induced by the carcinogen N-methyl-N-nitrosourea (MNU) and tumor promotion can be stimulated by the administration of testosterone, combinated or not with the use of pesticides. The present study aimed to evaluate the influence of DDVP on the development of morphological alterations in the ventral prostate of rats, after chemical induction by MNU. Thereunto, 32 Fischer 344 rats, aged 90 days, were randomly separated into four experimental groups: Control, DDVP, MNU, MNU+DDVP. For chemical induction, the MNU and MNU+DDVP groups were inoculated with MNU (15 mg/kg), followed by daily subcutaneous injections of testosterone cypionate (2.5 mg/kg), for 20 days. Animals of the DDVP and MNU+DDVP groups, from 120 to 240 days old, received the basal diet supplemented with 10 mg/kg of DDVP. Subsequent to the experimental period, the prostatic ventral lobe of each animal was collected and subjected to morphometric-stereological and histopathological analysis, for the classification of the lesions found. The incidence of alterations in epithelium and stroma of all experimental groups was evaluated, such as epithelial hyperplasia (increased cell population with stratification, without atypia) (Figure A and B); atrophy (retraction of the epithelium with reduction of the secretory cytoplasm) (Figure C and D); periacinary inflammatory foci (presence of inflammatory cell infiltrate) (Figure E and F); cell atypia with presence of cytoplasmic inclusions (Figure G and H) and metaplasia with stromal hyperplasia (change in the pattern of the original epithelium without proliferative change, with an increase in the adjacent stroma) (Figure I to L). The results showed a significant increase of epithelial hyperplasia areas in MNU and MNU+DDVP groups, with 100% of the animals showing this lesion (Table 1), which demonstrates a relation between chemical induction and the proliferative process. The morphometric-stereological analysis confirmed this finding, revealing a relatively greater volume of the epithelial compartment in MNU+DDVP, compared to the Control group (Table 2). Thus, it can be concluded that DDVP, even in low concentration, associated with chemical induction by MNU, was able to disturb the morphology of the ventral prostate, promoting proliferative epithelial lesions, strongly associated with the process of carcinogenesis.

Abstract 1146: Development of CRISPR human Skp2 knock-in in the prostate of mouse and the associated prostate organoids for testing Skp2 targeting agents

Abstract S-phase kinase associated protein 2 (Skp2) is a promising drug target as studies have demonstrated that Skp2 is required for spontaneous tumor development that occurs in the retinoblastoma protein (pRb), Pten or p53 deficient mice. We therefore have aimed to establish CRISPR human Skp2 (hSkp2) knock-in in the prostate of mouse to examine its role in prostate carcinogenesis. In addition, Skp2 overexpressing organoids are being developed for convenient and efficient screening of specific Skp2 inhibitors. cDNA of hSkp2 and porcine teschovirus-1 (P2A) was introduced into immediate upstream of mouse probasin coding sequencing using CRISPR knock-in targeting approach to establish transgenic mice specifically overexpressing hSkp2 in the prostate glands. Quantitative PCR, western blot and immunohistochemistry (IHC) were used to identify the expression levels of Skp2 and histopathological analyses were performed after H&amp;E staining. Prostate glands were dissected and digested for organoid culture, then morphological features and viability of organoids were evaluated following treatments of potential Skp2 inhibitors. Prostate glands and other tissues from mice of three transgenic lines which were genotyped positive were dissected and hSkp2 was detected in ventral, dorsal and lateral prostate lobes, but not in testis, kidney, spleen, and liver. Line 10092 was found to have the highest expression of hSkp2 among these three lines. Prostate weight slightly increased in transgenic mice compared to that of wild-type control mice. Hyperplasia, prostatic intraepithelial neoplasia (PIN), and carcinoma were noted, which suggested that overexpression of hSkp2 in the prostate of mice promoted proliferation and prostate tumorigenesis. Organoids that were derived from the prostate of hSkp2 transgenic mice recapitulated prostatic phenotypic features and maintained a high level of hSkp2. A novel Flavokawain A derivative was found to selectively alter the morphology of hSkp2 organoids and decrease the viability. Prostatic lesions found in the hSkp2 knock-in mice demonstrated a potential role of hSkp2 in early prostate carcinogenesis and organoids overexpressing hSkp2 have the utility for screening out compounds targeting Skp2 in prostate cancer. Citation Format: Liankun Song, Shan Xu, Kia Arabzadehkaffash, Ali Fazelpour, Dongjun Fu, Matthew Tippin, Xiaolin Zi. Development of CRISPR human Skp2 knock-in in the prostate of mouse and the associated prostate organoids for testing Skp2 targeting agents [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 1146.

Effect of Allium Cepa Seed Extracts on the Histopathological Changes of Ventral Prostatic Lobe in Diabetic Rats

Effect of Allium Cepa Seed Extracts on the Histopathological Changes of Ventral Prostatic Lobe in Diabetic Rats

Morphological and Biochemical Characteristics of Prostate Hyperplasia during Sulpiride Treatment.

We studied morphological changes in the prostate ventral lobe, proliferative activity of the epithelium in prostate acini, and the levels of prolactin and prostate-specific antigen in the blood serum of Sprague-Dawley rats after repeated injections of sulpiride in a dose of 40 mg/ kg over 30 and 60 days and in 10 and 30 days after withdrawal. Morphological and morphometrical analysis of hyperplastic changes in the prostate ventral lobe was performed. Ki-67+ proliferating epithelial cells in the acini were counted. The dynamics of serum concentrations of prolactin and prostate-specific antigen was evaluated by ELISA. Morphological and morphometrical analysis and evaluation of the content of Ki-67+ cells demonstrated epithelium hyperplasia in the prostate ventral lobe after sulpiride treatment for 30 or 60 days and in 10 days after withdrawal, but serum level of prostate-specific antigen did not differ from the control. After 60-day sulpiride treatment and in 30 days after withdrawal, pronounced hyperplastic changes of prostate and elevated concentrations of prostate-specific antigen (but not prolactin) were observed. Thus, administration of sulpiride (40 mg/kg) to Sprague-Dawley rats for 60 days allows, by morphological criteria and serum level of prostate-specific antigen, to model stable hyperplastic changes in the prostate corresponding to benign prostatic hyperplasia in humans.

Ultrasonographic Follow-up of the Multistep Protocol for Prostate Cancer Induction in Wistar Rats.

This work intended to improve the knowledge of the rat model of prostate cancer (PCa) by ultrasonographic monitoring. Male Wistar rats were divided into control (n=8) and PCa (n=14) groups. PCa development was induced in the PCa group through the sequential administration of the anti-androgenic drug flutamide, testosterone propionate and the carcinogenic N-methyl-N-nitrosourea. The prostate was evaluated by ultrasonography at five timepoints along 49 weeks of the experimental protocol. Ventral prostate lobes were observed in all ultrasonographic examinations. The ventral prostate area of the control group increased gradually between the first and the last ultrasonographic examination. The ventral prostate area of PCa groups decreased due to flutamide administration and increased after androgen and carcinogen administration. The area of the dorsal prostate lobe increased between the fourth and the fifth ultrasonographic examination. In the last ultrasonographic examination, hypoechoic and anechoic lesions were observed in the PCa group. To our knowledge, this is the first study presenting a follow-up of rat prostatic dimensions by ultrasonography. Ultrasonography is a feasible approach for prostate cancer monitoring in experimental models.

Abstract 3591: Genetic and pharmacological inhibition of fatty acid synthase (FASN) attenuates prostate cancer driven by Pten loss

Abstract Inactivation of the tumor suppressor gene phosphatase and tensin homologue (PTEN) is a common alteration in prostate cancer, occurring in the majority of metastatic castration-resistant prostate cancer (mCRPC) cases, and is associated with a more aggressive phenotype and worse prognosis. Another hallmark of prostate cancer is the dysregulation of lipid metabolism with overexpression of fatty acid synthase (FASN), the key enzyme in the de novo synthesis of fatty acids. FASN activity is essential for the generation of new cellular membrane lipids and signaling molecules, conferring growth and survival advantages for cells that overexpress it. Increased FASN expression enhances the aggressive phenotype associated with PTEN loss. To study how inhibition of FASN affects prostate carcinogenesis, we developed a mouse model combining prostate-specific homozygous knockout (KO) of Pten and Fasn genes. Deletion of Fasn, Pten or Fasn/Pten was achieved by expressing Cre recombinase under the control of a Probasin promoter, generating different genotypes: PtenloxP/loxPFasnwt/wtCrepos (P-KO), Ptenwt/wtFasnloxP/loxPCrepos (F-KO) and PtenloxP/loxPFasnloxP/loxPCrepos (F/P-dKO). Inhibition of FASN decreased the weight and volume of the ventral and anterior lobes of murine prostate when compared with Pten single KO mice. Histopathological analysis of the tissues also revealed a reduction in reactive stroma in the ventral and anterior lobes of F/P-dKO in comparison to the P-KO group. This phenotype is suggestive of inhibition of microinvasion. To assess whether FASN inhibition impairs cancer aggressiveness, we tested a novel FASN inhibitor compound, IPI-9119 (Infinity Pharmaceuticals), in epithelial cells from prostate of mice heterozygous for Pten loss. Pharmacological inhibition of de novo lipogenesis lead to decreased cell proliferation and reduction in invasion and motility. Finally, combined loss of PTEN with FASN overexpression assessed in 660 prostate cancer patients with 14.2 years of median follow up, was associated with lethality. Taken together, these data suggest that endogenous lipogenesis supports invasion in a PTEN null background. Since de novo lipogenesis contributes to the aggressive phenotype induced by PTEN loss in murine prostate, targeting lipid metabolism may represent an attractive therapeutic strategy in the context of prostate cancer driven by PTEN loss. Citation Format: Caroline F. Ribeiro, Débora C. Bastos, Hubert Pakula, Thomas Ahearn, Jéssica Nascimento, John S. Clohessy, Lorelei Mucci, Silvio M. Zanata, Giorgia Zadra, Massimo Loda. Genetic and pharmacological inhibition of fatty acid synthase (FASN) attenuates prostate cancer driven by Pten loss [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 3591.

The dynamic change of gene expression profiles of rats’ benign prostate hyperplasia tissues after denervation of sympathetic nerve

Objective To investigate the dynamic change of gene expression profiles of rats’ benign prostate hyperplasia (BPH) tissues after denervation of sympathetic nerve. Methods Twelve spontaneous hypertension rats (SHR), male, 29 weeks old, 3 for normal control (N), 9 were cut the adrenergic inferior epigastric innervation to prostate. Three N, 3 SHR 3 days after operation (D3), 3 D11 and 3 D21 were sacrificed and collected the ventral lobe of prostate for histopathological examination, RNA extraction, gene expression profiles microarray, real-time quantitative polymerase chain reaction (Real-time PCR) and bioinformatics analysis. Results The smooth muscle in rats’ BPH tissues presented a transient structure and function damage after losing the sympathetic innervations. Gene expression profiles microarray tests were finished successfully, their reliability were verified by the real-time PCR. Clustering analysis indicated that the number of up-regulated gene and down-regulated gene in gene set D3/N were 318, 117, D11/N were 192, 66 and D21/N were 44, 92, respectively. GO function and Pathway enrichment analysis indicated that these differentially expressed genes were involved in numerous molecular function, biological progress, cellular compartment and signal pathway, the numbers also decreased with the extension of time. The GO function and signal pathway were related with inflammation, immune response and cellular response to injure and stress at early stages, related with repairmen of tissue compartment and cellular function in late period. Most of the top ranked signal pathways at early stages had relationship with the activation of complement system, and genes involved were up-regulated. Conclusion The impact of denervation of sympathetic nerve for rats’ BPH tissue (mainly smooth muscle) in molecular level also is transient, and the abnormal activation of complement system seems to play a important role in the serial molecular events. Key words: Benign prostate hyperplasia; Denervation; Sympathetic innervations; Gene expression profile; Bioinformatics analysis; Rat

The potential protective role of flaxseed extract on ventral prostate in a rat model of streptozotocin-induced diabetes: A histological and immunohistochemical study

Background: Diabetes mellitus is a chronic disorder that can affect the reproductive system leading to fertility dysfunction. The prostate plays an important role in the reproduction and it is a frequent target for diabetic complications. Flaxseed is a promising alternative in many chronic diseases due to its health benefits. It has a proposed role in controlling diabetes due to its wide pharmacological activities. im: Evaluation of the possible protective role of flaxseed extract on the ventral prostate gland of diabetic rats. Materials & Methods: Fifty adult male albino rats were used as a control group and experimental group that received a single intra-peritoneal injection of streptozotocin (60 mg/kg). The experimental rats with confirmed diabetes mellitus were subdivided into: diabetic group that kept without treatment and diabetic–flaxseed group (400 mg/kg/day orally). After four weeks, specimens of the ventral prostate lobes were processed for histological and immunohistochemical study using antibodies against Bcl-2, α-smooth muscle actin and androgen receptor. Results: In comparison with the control, specimens of the diabetic group showed glandular epithelial atrophy with thickening of the fibromuscular stroma. Reduced epithelial height and a statistically significant decrease in Bcl2 immunoexpression were observed in the epithelial cells. A statistically significant increase in the collagen fibers and α-SMA immunoexpression was seen in the stroma. Androgen receptor immunoreactivity was significantly decreased in both epithelial and stromal cells. In contrast, minimal changes appeared in diabetic-flaxseed group that received flaxseed extract. Conclusion: Diabetes induced structural alterations in the ventral prostate gland of rats. Administration of flaxseed extract attenuated these effects and partially preserved the glandular structure.

Growth hormone-releasing hormone antagonists reduce prostatic enlargement and inflammation in carrageenan-induced chronic prostatitis.

Inflammation plays a key role in the etiology of benign prostatic hyperplasia (BPH) through multiple pathways involving the stimulation of proliferation by cytokines and growth factors as well as the induction of the focal occurrence of epithelial-to-mesenchymal transition (EMT). We have previously reported that GHRH acts as a prostatic growth factor in experimental BPH and in autoimmune prostatitis models and its blockade with GHRH antagonists offer therapeutic approaches for these conditions. Our current study was aimed at the investigation of the beneficial effects of GHRH antagonists in λ-carrageenan-induced chronic prostatitis and at probing the downstream molecular pathways that are implicated in GHRH signaling. To demonstrate the complications triggered by recurrent/chronic prostatic inflammation in Sprague-Dawley rats, 50 μL 3% carrageenan was injected into both ventral prostate lobes two times, 3 weeks apart. GHRH antagonist, MIA-690, was administered 5 days after the second intraprostatic injection at 20 μg daily dose for 4 weeks. GHRH-induced signaling events were identified in BPH-1 and in primary prostate epithelial (PrEp) cells at 5, 15, 30, and 60 min with Western blot. Inflammation induced prostatic enlargement and increased the area of the stromal compartment whereas treatment with the GHRH antagonist significantly reduced these effects. This beneficial activity was consistent with a decrease in prostatic GHRH, inflammatory marker COX-2, growth factor IGF-1 and inflammatory and EMT marker TGF-β1 protein levels and the expression of multiple genes related to EMT. In vitro, GHRH stimulated multiple pathways involved in inflammation and growth in both BPH-1 and PrEp cells including NFκB p65, AKT, ERK1/2, EGFR, STAT3 and increased the levels of TGF-β1 and Snail/Slug. Most interestingly, GHRH also stimulated the transactivation of the IGF receptor. The study demonstrates that GHRH antagonists could be beneficial for the treatment of prostatic inflammation and BPH in part by inhibiting the growth-promoting and inflammatory effects of locally produced GHRH.