15 research outputs found

    Mmp-2 And Mmp-9 activities and Timp-1 and Timp-2 expression in the prostatic tissue of two ethanol-preferring rat models

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    We investigated whether chronic ethanol intake is capable of altering the MMP-2 and MMP-9 activities and TIMP-2 and TIMP-1 expression in the dorsal and lateral prostatic lobes of low (UChA) and high (UChB) ethanol-preferring rats. MMP-2 and MMP9 activities and TIMP-1 and TIMP-2 expression were significantly reduced in the lateral prostatic lobe of the ethanol drinking animals. Dorsal prostatic lobe was less affected showing no significant alterations in these proteins, except for a reduction in the TIMP-1 expression in UChA rats. These important findings demonstrate that chronic ethanol intake impairs the physiological balance of the prostate extracellular matrix turnover, through downregulation of MMPs, which may contribute to the development of prostatic diseases. Furthermore, since these proteins are also components of prostate secretion, the negative impact of chronic ethanol intake on fertility may also involve reduction of MMPs and TIMPs in the seminal fluid2015COORDENA√á√ÉO DE APERFEI√áOAMENTO DE PESSOAL DE N√ćVEL SUPERIOR - CAPESFUNDA√á√ÉO DE AMPARO √Ä PESQUISA DO ESTADO DE S√ÉO PAULO - FAPESPsem informa√ß√£o2011/03394-4; 2011/13713-

    Fibrosis-related gene expression in the prostate is modulated by doxazosin treatment

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    Aims To gain new insights into the molecular mechanisms of action of doxazosin, we investigated the prostatic stroma ultrastructure and the expression of genes involved with fibrosis, such as collagen type I and III (COL1A1 and COL3A1, respectively) and TGF-beta 1, in the rat ventral prostate. Main methods Adult Wistar rats were treated with doxazosin (25 mg/kg/day), and the ventral prostates were excised at 7 and 30 days after treatment. Untreated rats were controls. Ventral prostates were subjected to ultrastructural, immunohistochemical, biochemical and molecular analyses. Key findings Doxazosin-treated prostates showed thickened bundles of collagen fibrils, activated fibroblasts, enlarged neurotransmitter vesicles and increased tissue immunostaining for collagen type I and type III when compared to untreated prostates. After 7 and 30 days of doxazosin treatment mRNA expression of COL1A1 and COL3A1 was significantly increased and reduced, respectively, compared to the control group. TGF-beta 1 mRNA and protein levels were increased after 7 days of doxazosin treatment, whereas only mRNA levels remained increased after 30 days of treatment. Significance Our data suggest that relaxation of smooth muscle cells by alpha-blockers interferes with the mechanical dynamics of the prostatic stroma extracellular matrix components, generating a pro-fibrotic effect probably via the TGF-beta 1 signaling pathway. Long term treatment with doxazosin may also lead to a reduced turnover of extracellular matrix components. Our results add to a better understanding of the molecular mechanisms behind the effects of alpha-blockade on prostatic histoarchitecture and the response to treatment for benign prostatic hyperplasia9125-2612811287CONSELHO NACIONAL DE DESENVOLVIMENTO CIENT√ćFICO E TECNOL√ďGICO - CNPQCOORDENA√á√ÉO DE APERFEI√áOAMENTO DE PESSOAL DE N√ćVEL SUPERIOR - CAPESFUNDA√á√ÉO DE AMPARO √Ä PESQUISA DO ESTADO DE S√ÉO PAULO - FAPESPsem informa√ß√£osem informa√ß√£o06/60114-6; 06/60115-2The authors thank the Electron Microscopy Center (Institute of Biosciences, UNESP, Botucatu, SP) for the transmission electron microscopy protocol procedures and for the use of Phillips electron microscope. This work was funded by FAPESP (S√£o Paulo State Research Foundation; grants 06/60114-6, to S.L.F. and 06/60115-2, to F.K.D); by FUNDUNESP (Foundation for Development of Univ Estadual Paulista), by CNPq, and by CAPES do Brazil. This paper represents part of the PhD thesis presented by F.K.D. to the University of Campinas‚ÄĒUNICAMP, Brazi

    Prostate telocytes change their phenotype in response to castration or testosterone replacement

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    Telocytes are CD34-positive cells with a fusiform cell body and long, thin cytoplasmic projections called telopodes. These cells were detected in the stroma of various organs, including the prostate. The prostate is a complex gland capable of undergoing involution due to low testosterone levels; and this condition can be reversed with testosterone replacement. Telocyte function in the mature prostate remains to be dermined, and it is not known whether telocytes can take place in tissue remodeling during prostate involution and regrowth. The present study employed structural, ultrastructural and immunohistochemical methods to investigate the telocyte's phenotypes in the ventral prostate (VP) from control (CT), castrated (CS) and testosterone replacement (TR) groups of adult male Wistar rats. Telocytes were found in the subepithelial, perimuscular and interstitical regions around glandular acini. Telocytes from CT animals have condensed chromatin and long and thin telopodes. In CS group, telocytes appeared quiescent and exhibited layers of folded up telopodes. After TR, telocytes presented loose chromatin, abundant rough endoplasmic reticulum and enlarged telopodes, closely associated with bundles of collagen fibrils. We called these cells "telocytes with a synthetic phenotype". As testosterone levels and glandular morphology returned toward to the CT group parameters, after 10 days ofTR, these telocytes progressively switched to the normal phenotype. Our results demonstrate that telocytes exhibit phenotypic plasticity upon androgen manipulation and interact with fibroblast and smooth muscle cells to maintain glandular architecture in control animals and during tissue remodeling after hormonal manipulation9CONSELHO NACIONAL DE DESENVOLVIMENTO CIENT√ćFICO E TECNOL√ďGICO - CNPQFUNDA√á√ÉO DE AMPARO √Ä PESQUISA DO ESTADO DE S√ÉO PAULO - FAPESP305391/2014-3; 306900/2016-5; 305840/2015-0; 310663/2018-02002/11102-4; 2014/26660-0; 2017/01063-

    Finasteride effects on epithelium-stroma interaction in the different rat prostatic lobes

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    Orientador: Sergio Luis FelisbinoDisserta√ß√£o (mestrado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: As c√©lulas do c√Ęncer prost√°tico s√£o geralmente dependentes, para o seu crescimento e sobreviv√™ncia, do est√≠mulo androg√™nico mediado pelos receptores de andr√≥geno (RA). Conseq√ľentemente, a manipula√ß√£o hormonal, como a castra√ß√£o e/ou o uso de antagonistas de RA, resulta na regress√£o do c√Ęncer. Entretanto, esses tratamentos raramente levam √† cura da doen√ßa em est√°gios mais avan√ßados e o c√Ęncer, conseq√ľentemente, torna-se independente de andr√≥geno. Importantes evid√™ncias sustentam a hip√≥tese de que as metaloproteinases de matriz (MMPs) t√™m papel fundamental nos m√ļltiplos passos da progress√£o tumoral, incluindo a promo√ß√£o tumoral, angiog√™nese, invas√£o e met√°stase. As MMPs constituem uma ampla fam√≠lia de endopeptidases dependentes de zinco e c√°lcio e atuam na degrada√ß√£o da matrix extracelular e da membrana basal. Dois importantes membros da fam√≠lia das MMPs, MMP-2 e MMP-9, pertencentes √† subfam√≠lia das gelatinases, s√£o conhecidas por degradarem especificamente o col√°geno do tipo IV, principal componente das membranas basais. Recentemente, o PCPT (Prostate Cancer Prevention Trial) mostrou que a finasterida, um inibidor da enzima 5_-redutase amplamente usado no tratamento da hiperplasia prost√°tica benigna, diminuiu significativamente a incid√™ncia do c√Ęncer prost√°tico quando comparado ao grupo que recebeu apenas placebo. Por√©m, tumores com grada√ß√£o de Gleason entre 7 e 10 foram significativamente mais comuns no grupo tratado com finasterida que no grupo placebo. Nesse sentido, o papel da finasterida na quimiopreven√ß√£o do c√Ęncer prost√°tico necessita de mais investiga√ß√£o. Assim, este trabalho examinou se as altera√ß√Ķes induzidas pela finasterida na morfologia e na fisiologia prost√°ticas t√™m impacto na express√£o e atividade das MMPs 2 e 9. Ratos machos Wistar (90 dias de idade) foram tratados diariamente com inje√ß√Ķes subcut√Ęneas de finasterida (25mg/Kg) durante 3, 7, 30 e 90 dias. Um grupo de animais controle recebeu diariamente inje√ß√Ķes subcut√Ęneas do ve√≠culo farmacol√≥gico (√≥leo de milho). Os lobos ventrais (PVs), dorsolaterais (PDLs) e anteriores (PAs) foram dissecados, pesados e processados para as an√°lises citoqu√≠micas, imunocitoqu√≠micas, morfom√©trico-estereol√≥gicas e de zimografias. A finasterida promoveu involu√ß√£o progressiva no compartimento epitelial e luminal, redu√ß√£o na altura e na prolifera√ß√£o das c√©lulas epiteliais e aumentou o √≠ndice apopt√≥tico e a √°rea estromal dos tr√™s lobos prost√°ticos. A an√°lise das zimografias mostrou que o tratamento com finasterida altera de modo lobo espec√≠fico o padr√£o gelatinol√≠tico da pr√≥stata do rato, com o aumento da pr√≥- MMP-9 no dia 30 e da forma ativa da MMP-9 nos dias 30 e 90 na PV, aumento da pr√≥- MMP-9 na PDL nos dias 7 e 30 e o aumento da forma intermedi√°ria da MMP-2 na PA no dia 7. Por√©m, a atrofia induzida pela finasterida n√£o promoveu um aumento significativo na atividade das MMPs, em contraste com o que √© encontrado na abla√ß√£o hormonal pela castra√ß√£o. Isso se deve, provavelmente, ao fato das altera√ß√Ķes promovidas pela finasterida na pr√≥stata constitu√≠rem um processo mais lento em compara√ß√£o com a castra√ß√£o. Assim, os nossos resultados sugerem que a finasterida √© eficaz no tratamento das afec√ß√Ķes prost√°ticas sem promover um ambiente favor√°vel para o crescimento de tumores invasivos e met√°staseAbstract: Prostate cancer cells are generally dependent on androgen stimulation mediated by the androgen receptor (AR) for growth and survival. Therefore, hormonal manipulation, such as castration and/or the use of AR antagonists, results in a regression of the cancer. However, these treatments very rarely lead to the ''cure¬Ņ¬Ņ of advanced disease and cancer eventually become androgen-independent. A substantial amount of evidence supports the hypothesis that Matrix Metalloproteinases (MMPs) play a key role in multiple steps of tumor progression, including tumor promotion, angiogenesis, invasion, and metastasis. MMPs constitute a broad family of zinc-binding endopeptidases that play a key role in degradation of the extracellular matrix and basement membrane. Two important members of the MMP family, MMP-2 and MMP-9, belonging to the gelatinase subfamily, are known to specifically cleave type IV collagen, the major component of basement membranes. Recently, The Prostate Cancer Prevention Trial (PCPT) showed that finasteride, a 5 alphareductase inhibitor widely employed in the treatment of benign prostatic hyperplasia, significantly decreased the incidence of prostate cancer versus placebo. However, Gleason score 7-10 tumors were significantly more common in the finasteride versus the placebo group. In this sense, the role of finasteride for prostate cancer chemoprevention needs further examination. Thus, this study examined whether the finasteride-induced changes on prostate morphology and phisiology has an impact on MMP-2 and MMP-9 expression and activity. Male Wistar rats (90 days of age) were treated daily with finasteride subcutaneously injected (25mg/Kg) during 3, 7, 30 and 90 days. A control group of animals received daily doses of vehicle (corn oil). The ventral (VPs), dorsolaterals (DLPs) and anterior (APs) lobes had been dissected, weighed and processed for citochemistry, immunocitochemistry, morphometric-stereological, and zymography analyses. Finasteride promoted a progressive involution in the epithelium and lumen compartments, reduced of the epithelial cell heights and proliferation and increased apoptosis and the stromal area in the three prostatic lobes. Gelatin zymography analysis showed that finasteride treatment changed the gelatinolitic profile in the prostatic lobes, with an increase in the pro-MMP-9 at day 30 and in the active-MMP-9 at days 30 and 90 in VP, increase in the pro-MMP-9 inDLP at days 7 and 30 and an increase in the intermediate-MMP-2 in PA at day 7. However, finasteride induced-atrophy did not cause a significant increase in MMPs activity, in contrast with is found in androgen ablation by castration. This is probably due to the fact that finasteride changes in prostate are a slow process in comparison with castration. Take together, our results suggest that finasteride is effective in treating prostate diseases without creating a most favorable environment for tumor growth invasion and metastasisMestradoBiologia CelularMestre em Biologia Celular e Estrutura

    The Anti-Inflammatory Effects of the Methanolic Extract and Fractions from Davilla elliptica St. Hil. (Dilleniaceae) on Bothrops jararaca Envenomation

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    Inflammation and haemorrhage are the main characteristics of tissue injury in botropic envenomation. Although some studies have shown that anti-venom prevents systemic reactions, it is not efficient in preventing tissue injury at the site of the bite. Therefore, this work was undertaken to investigate the anti-inflammatory effects of the methanolic extract and fractions from D. elliptica and to evaluate the role of matrix metalloproteinases (MMPs) in this process. Effects of the extract and fractions from D. elliptica were evaluated using a carrageenan-induced paw oedema model in rats, and leukocyte rolling was visualized by intravital. The quantification of MMPs activities (MMP-2 and MMP-9) extracted from the dermis of mice treated with extract and fractions alone or incubated with venom was determined by zymographic analyses. Our results show that intraperitoneal (i.p.) injection of fractions significantly reduced paw oedema after the carrageenan challenge. Treatment with the tannins fraction also resulted in considerable inhibition of the rolling of leukocytes and this fraction was able to decrease the activation of MMP-9. These results confirmed the anti-inflammatory activity of the methanolic extract and tannins fraction of D. elliptica and showed that the dermonecrosis properties of B. jararaca venom might be mediated through the inhibition of MMP-9 activity.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

    Cytoxicity and Apoptotic Mechanism of Ruthenium(II) Amino Acid Complexes in Sarcoma-180 Tumor Cells

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    Over the past several decades, much attention has been focused on ruthenium complexes in antitumor therapy. Ruthenium is a transition metal that possesses several advantages for rational antitumor drug design and biological applications. In the present study, five ruthenium complexes containing amino acids were studied in vitro to determine their biological activity against sarcoma-180 tumor cells. The cytotoxicity of the complexes was evaluated by an MTT assay, and their mechanism of action was investigated. The results demonstrated that the five complexes inhibited the growth of the S180 tumor cell line, with IC50 values ranging from 22.53 mu M to 50.18 mu M, and showed low cytotoxicity against normal L929 fibroblast cells. Flow cytometric analysis revealed that the [Ru(gly)(bipy)(dppb)] PF6 complex (2) inhibited the growth of the tumor cells by inducing apoptosis, as evidenced by an increased number of Annexin V-positive cells and G0/G1 phase cell cycle arrest. Further investigation showed that complex 2 caused a loss of mitochondrial membrane potential; activated caspases 3, caspase-8, and caspase-9 and caused a change in the mRNA expression levels of caspase 3, caspase-9 as well as the bax genes. The levels of the pro-apoptotic Bcl-2 family protein Bak were increased. Thus, we demonstrated that ruthenium amino acid complexes are promising drugs against S180 tumor cells, and we recommend further investigations of their role as chemotherapeutic agents for sarcomas.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

    Terminalia catappa L.: A medicinal plant from the Caribbean pharmacopeia with anti-Helicobacter pylori and antiulcer action in experimental rodent models

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    Ethnopharmacological relevance: Terminalia catappa L. (Combretaceae) is a medicinal plant listed as a pharmacopeia vegetable from Caribbean to treat gastritis. The objective of this study was to evaluate the gastroprotective and healing effect of the aqueous fraction (FrAq) obtained from the leaves of Terminalia catappa and to determine the antiulcer mechanism of action in experimental rodent models and its activity to Helicobacter pylori.Material and methods: In rodents, the FrAq was challenged by different necrotizing agents, such as absolute ethanol and ischemia-reperfusion injury. The antiulcer mechanism of action of FrAq was assessed and the healing effects of the fraction after seven and 14 days of treatment was evaluated by matrix metalloproteinase activity (MMP-2 and MMP-9). The toxicological effect of subacute treatment with FrAq during 14 days of treatment was also analyzed. The anti-Helicobacter pylori activity was determined by microdilution. The phytochemical study of the fraction was analyzed by experiments with FIA-ESI-IT-MSn (Direct Flow Analysis-ionization Electrospray Ion Trap Tandem Mass Spectrometry) and high performance liquid chromatography (HPLC) coupled to a photodiode array (PDA).Results: Oral treatment with FrAq (25 mg/kg) significantly decreased the number of ulcerative lesions induced by ethanol and ischemia/reperfusion injury. The action of FrAq was mediated by the activation of defensive mucosa-protective factors, such as increases in mucus production, the nitric oxide (NO) pathway and endogenous prostaglandins. Oral treatment with FrAq for seven and 14 days significantly reduced the lesion area (80% and 37%, respectively) compared to the negative control group. Analyses of MMP-9 and MMP-2 activity from gastric mucosa confirmed the accelerated gastric healing effect of FrAq. This extract also presented considerable activity against Helicobacter pylori. The mass spectrum and MS/MS of the aqueous fraction indicates the existence of many different phenolic compounds, including punicalagin, punicalin, and gallagic acid, among others.Conclusions: We concluded that FrAq from Terminalia catappa leaves has excellent preventive and curative effects on acute and chronic induced gastric ulcers and showed an important profile against Helicobacter pylori. (C) 2014 Elsevier Ireland Ltd. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES

    Finasteride and doxazosin effects on rat prostate : ultrastructural and type Iand type III and TGF-1 expression analyses

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    Orientador: S√©rgio Luis FelisbinoTese (doutorado) - Universidade Estadual de Campinas, Instituto de BiologiaResumo: A finasterida e a doxazosina s√£o drogas usadas no tratamento da hiperplasia prost√°tica benigna (HPB) e, mais recentemente, est√£o sendo tamb√©m usadas na quimiopreven√ß√£o do c√Ęncer de pr√≥stata (CaP), principalmente devido aos seus efeitos na indu√ß√£o de apoptose das c√©lulas epiteliais prost√°ticas. Entretanto, pouca aten√ß√£o tem sido dispensada aos efeitos destes f√°rmacos sobre o estroma glandular. Assim, o objetivo deste estudo foi analisar os efeitos da finasterida e da doxazosina no arranjo estromal e em componentes espec√≠ficos, como o col√°geno do tipo I, col√°geno do tipo III e o TGF-Beta 1. Para isso, foram empregadas t√©cnicas morfol√≥gicas, bioqu√≠micas e moleculares. O tratamento com finasterida promoveu apoptose epitelial, altera√ß√Ķes na membrana basal epitelial e na membrana basal das c√©lulas musculares lisas, al√©m da diminui√ß√£o na express√£o dos col√°genos I e III. O bloqueio ?-adren√©rgico com doxazosina aumentou a quantidade de fibras do sistema el√°stico, promoveu apoptose nas c√©lulas epiteliais, al√©m de alterar a express√£o dos col√°genos I e III. Os dois f√°rmacos influenciaram o aumento da express√£o do TGF-Beta 1, que parece estar mais relacionado com o evento da apoptose no tratamento com a finasterida e com as altera√ß√Ķes das fibras col√°genas e a ativa√ß√£o de fibroblastos no tratamento com a doxazosina. Conclui-se que as altera√ß√Ķes estromais p√≥s tratamento com finasterida e doxazosina podem contribuir para a regress√£o prost√°tica esperada no tratamento da HPB e na ruptura de eventos par√°crinos respons√°veis pela evolu√ß√£o tumoral no CaPAbstract: Finasteride and doxazosin are drugs used in the benign prostatic hyperplasia (BPH) treatment and, more recently, they're have been used in the prostate cancer (PCa) chemoprevention, mainly because their apoptotic effect in the prostatic epithelial cells. However, few attentions have been given to the effects of these drugs in the glandular stroma. Thus, the objective of this study was to analyze the effects of the finasteride or doxazosin treatment on stroma arrangement and on specific components, like collagen type I, collagen type III and TGF-Beta 1. For this aim, we have used morphological, biochemical and molecular approaches. Finasteride treatment provoked epithelial apoptosis, epithelial basal membrane and smooth muscle cells basal membrane alterations, besides decrease the type I and type III collagen fibers expression. The alpha-adrenergic blockade increased the elastic fibers system, promoted apoptosis in epithelial cells, as well altered the type I and type III collagen fibers expression. Both drugs influenced the TGF-Beta 1 up-regulation that show to be related with apoptosis in finasteride treatment and with collagen fibers alterations and fibroblasts activation in doxazosin treatment. In conclusion, the stromal alterations promoted by finasteride and doxazosin treatments can contribute to the prostate regression expected in the BPH treatment and to the rupture of paracrine events responsible by tumoral evolution in the PCaDoutoradoBiologia CelularDoutor em Biologia Celular e Estrutura

    Caffeine reduces cadmium accumulation in the organism and enhances the levels of antioxidant protein expression in the epididymis

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    The aim of this study was to investigate the effects of caffeine (20. mg/L) intake on cadmium (15. mg/L) accumulation in the rat blood, testes, epididymis and prostate as well as cadmium-induced changes to the antioxidant defense system of the epididymis. Caffeine reduced the cadmium concentration in all tissues analyzed. Meanwhile, cadmium reduced catalase activity and increased superoxide dismutase (SOD) activity in the epididymis. Caffeine increased SOD activity, catalase and glutathione tissue expression and sustains the cadmium's effect on catalase and GSP-Px activity. No differences in the expression of metallothionein and lipid peroxidation were observed among the different treatments in the epididymis. In conclusion, low doses of cadmium alter the antioxidant enzymatic profile of the epididymis, but not induced oxidative lipid damage. Caffeine intake reduces overall cadmium accumulation in the organism and enhances the levels of antioxidant protein expression in the epididymis, thus exerting a protective effect against this metal. © 2012 Elsevier Inc
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