INTRODUÇÃO: A urticária crônica igmaidiopática (UCI) é uma doença desencadeada pela desgranulação de basófilos e mastócitos com consequente liberação de histamina, sendo que o perfil imunológico nesta doença não é bem estabelecido. As estatinas, inibidores da 3-hidroxi-3-metilglutaril coenzima A redutase, apresentam efeitos antiinflamatórios e imunomodulatórios. O efeito desta droga tem sido estudado em muitas doenças inflamatórias crônicas, incluindo doenças autoimunes, mas não existem evidências na UCI. OBJETIVOS: O objetivo deste estudo foi analisar o efeito das estatinas na resposta imune e sua a influência na expressão de genes regulatórios e relacionados com a resposta inflamatória. MÉTODOS: A resposta limfoproliferativa a mitógenos e antígeno-específica de 22 pacientes com UCI e 41 controles na presença de estatinas (0,25-25 µM) foi analisada pela incorporação de timidina após 3 ou 6 dias de cultura. A progressão do ciclo celular e apoptose foi realizada pela incorporação de bromodeoxiuridina (Brdu) ao DNA após estímulo por PHA ou PWM e analisada por citometria de fluxo. A secreção de citocinas foi quantificada por ELISA e a expressão de mRNA de fatores regulatórios e pró-inflamatórios quantificados por real-time PCR. RESULTADOS: Os resultados evidenciaram que as estatinas em elevadas concentrações são capazes de inibir a capacidade mitogênica das células T e B seja dos indivíduos saudáveis ou de pacientes com UCI. A inibição da proliferação celular mediada pelas estatinas foi decorrente ao bloqueio na etapa inicial do ciclo celular (Fase G0/1), o que impediu o prosseguimento para outras fases do ciclo (S e G2/M). A diminuição da resposta proliferativa em resposta a um mitógeno como a PHA resultou na inibição da ativação celular pela estatina e a significante redução na produção de citocinas como IFN-?, IL-10, IL-17A e IL-5. Em contraste, o efeito modulatório das estatinas ao estímulo com LPS inibiu a produção de TNF-? e MIP-1? pelas células dos controles, mas não influenciou na produção de citocinas pró-inflamatórias pelas CMN dos pacientes com UCI. Somente a incubação prévia das células com as drogas, em alta concentração (25µM), foi possível verificar a modulação negativa na produção de IL-6 e MIP1-? para ambos os grupos, mas não para o TNF-? para os pacientes. A sinvastatina foi capaz exercer efeito modulatório mais pronunciado que a lovastatina na produção de citocinas induzidas por LPS. Os resultados evidenciaram que os pacientes com UCI possuem uma diminuição da expressão da enzima IDO e aumento de SOCS3 nas CMN. A sinvastina não altera esse perfil e previne a expressão de fatores inflamatórios como RORC?t e NALP3 inflamassomas. CONCLUSÕES: Em conjunto, os resultados sugerem um desequilíbrio dos mecanismos regulatórios que poderiam contribuir com a cronicidade e o perfil inflamatório na UCI. As estatinas apresentam maior efeito antiinflamatório que pró-inflamatório, sugerindo ter potencial clínico para o tratamento de doenças crônicas como a UCI. / INTRODUCTION: Chronic Idiopathic Urticaria (CIU) is a disease triggered by degranulation of basophils and mast cells with consequent histamine release and the CIU immunological profile is not well established. Statins, 3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors, also display a broad immunomodulatory property. Statins have been studied in several chronic inflammatory diseases, including autoimmune disorders, but there are no evidences in CIU disease. OBJECTIVES: The aim of this study was to verify the effect of statins the immune response, and the expression of genes related to regulatory and inflammatory response focusing in CIU patients and healthy controls (HC). METHODS: Lymphoproliferative response to mitogens or recall antigens of 22 patients with CIU and 41 HC with statins (0,25-25µM) was analyzed by timidine incorporation after 3 or 6 days of cell cultures. Cell cycle progression and apoptosis were assessed by bromodeoxyiridine (BrDU) incorporation to DNA upon PHA or PWM stimulus by flow cytometry. Cytokines secretion was measured by ELISA and mRNA of regulatory and proinflammatory genes were analyzed by quantitative real-time PCR. RESULTS: The results showed that high concentrations of statins can inhibit the mitogenic capacity of T and B cells of HC or CIU patients. The inhibition of cell proliferation mediated by statins was due to blockage in the initial phase of the cell cycle (G0/1), which prevented progress to cycle phases (S and G2/M). The decreased proliferative response in response to PHA mediated by statin resulted in a significant inhibition of IFN-?, IL-10, IL-17A and IL-5 secretion levels. Statin effect in response to LPS showed inhibition of TNF-? and MIP-1? secretion by cells from HC, but did not influence the production by PBMC of CIU. It was necessary the pre-incubation of cells with drugs at high concentration (25µM) to verify the negative modulation of IL-6 and MIP1-? secretion in both groups, except for TNF-? in CIU. Simvastatin was able to exert more pronounced modulatory effect than lovastatin in cytokine production induced by LPS. Furthermore, CIU patients have a decreased expression of the enzyme IDO and increased of SOCS3 in PBMC, which were not modified by simvastatin, whereas prevented the upregulation of proinflammatory factor as RORC?t and NALP3 inflammasomes. CONCLUSIONS: Altogether, the results evidenced an imbalance of regulatory mechanisms that could contribute to chronic evolution and inflammatory profile in CIU. Statins exhibited more anti-inflammatory effects than proinflammatory, suggesting a potential clinical role for treatment in chronic diseases as CIU.

Orientador: André Almeida Schenka / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciências Médicas / Made available in DSpace on 2018-08-25T22:24:18Z (GMT). No. of bitstreams: 1 Renno_AndreLisboa_D.pdf: 23208237 bytes, checksum: c782ca447d3fee7cb5573e4b122b7f94 (MD5) Previous issue date: 2014 / Resumo: O câncer mamário é a neoplasia maligna mais incidente e a principal causa de óbito por malignidade no sexo feminino no Brasil e no mundo. Estipula-se que há mais de 1.2 milhões de novos casos anuais de câncer de mama, e que a heterogeneidade e a complexidade molecular do câncer de mama dificultam estratégias terapêuticas de prevenção e tratamento desta doença. Atualmente, acredita-se que, em diversas neoplasias, incluindo o câncer de mama, a célula alvo de mutações cumulativas responsáveis pelo desenvolvimento do fenótipo canceroso é uma célula-tronco adulta. Independentemente da origem da neoplasia (se em célula madura/diferenciada ou em CT), é possível constatar in vitro e in vivo, na grande maioria dos tumores malignos, uma subpopulação de células indiferenciadas, com características fenotípicas de célula-tronco. Tais células são designadas como "células tronco cancerosas ou neoplásicas (CTNs)". Com frequência, especula-se se as CTNs seriam responsáveis pela heterogeneidade morfológica e molecular de algumas neoplasias mamárias. Em conjunto, essas peculiaridades das CTNs as tornam importantes alvos no desenvolvimento de novas abordagens farmacoterapêuticas antineoplásicas. Recentemente, Gauthaman et al (2009) demonstraram de forma inédita em estudos in vitro que estatinas apresentam efeito inibitório específico sobre células tronco embrionárias com alterações cariotípicas e células de linhagens neoplásicas mamárias com fenótipo CTN, não afetando o crescimento de células tronco normais. As estatinas são inibidores competitivos da 3-hidroxi-3-metilglutaril coenzima A (HMG-CoA) e são amplamente utilizada para o tratamento de doenças cardiovascular primário e secundário. Além de amplamente utilizadas na prevenção e tratamento de doenças cardiovasculares secundárias a dislipidemias, evidências cumulativas apontam para um possível papel destas drogas na prevenção ou regressão de processos neoplásicos. Entre os efeitos antineoplásicos comprovados das estatinas, destacam-se: a inibição da proliferação celular, a promoção de apoptose, a inibição da angiogênese e a prevenção de metástases. Assim, buscou-se neste trabalho elucidar o efeito da sinvastatina e pravastatina sobre células progenitoras e CTNs e em algumas vias de sinalização intracelular em modelo de carcinogênese mamária (baseado na indução com 7,12 dimetilbenz(a)antraceno[DMBA]) em ratas Sprague-Dawley. Após um tratamento de 14 dias com as estatinas, as mamas das ratas foram analisadas para verificar a imunoexpressão de células progenitoras e CTNs (CD133, CD24, CD44 e EpCAM), variáveis biológicas (volume tumoral, mitose, índice proliferativo) além da análise proteica de Akt e Src. A maior dose da sinvastatina testada (40 mg/Kg) diminui o número de tumores desenvolvidos, volume e incremento tumoral e os índices de proliferação celular. Não houve alteração da percentagem de necrose com o tratamento com as estatinas. Ainda, sinvastatina diminuiu os níveis da fosforilação da Akt e aumento da PTEN, não havendo diferenças significantes nos níveis da Src. Sinvastatina também foi capaz de reduzir o número de células positivas CD133, CD24 e CD44. Pelas doses testadas, não houve diferença dos parâmetros biológicos analisados com o tratamento com a pravastatina. Em conclusão, neste modelo, o tratamento crônico com a sinvastatina apresentou efeitos citostáticos, ações reguladoras na via da Akt além do controle de células progenitoras e CTNs em modelo in vivo de carcinoma mamário / Abstract: Breast cancer is the malignant neoplasm with the highest incidence and the main cause of death by cancer within females in Brazil and in the world. It is estimated that there are over 1.2 million new annual cases of breast cancer. The heterogeneity and the molecular complexity of this type of cancer complicate the therapeutic strategies for its prevention and treatment. Nowadays, it is believed that in many different neoplasms, including breast cancer, the cell which is the target of cumulative mutations responsible for the development of the cancerous phenotype is an adult stem cell. Regardless the origin of the neoplasm (whether in mature/differentiated cell or in SC), a subpopulation of undifferentiated cells with phenotypic characteristics of stem cells can be seen in vitro and in vivo in most malignant tumours. These cells are designated as "neoplastics or cancer stem cells (CSCs)". It is often especulated whether CSCs would be responsible for the molecular and morphological heterogeneity in some breast neoplasms. The peculiarities of the CSCs make them a relevant/an important/a serious object for the development of new antineoplastic pharmacotherapeutic approaches. Recently, Gauthaman et al (2009) demonstrated in unprecedented in vitro studies that statins exhibit specific inhibitory effect on embryonic stem cells with karyotypic alterations and neoplastic mammary cell lines with phenotype CSC, not affecting the growth of normal stem cells. Statins are competitive inhibitors of coenzyme 3-hydroxy-3-methylglutaryl A (HMG-CoA) reductase and are widely used for the primary and secondary treatment of cardiovascular diseases. Moreover, cumulative evidence points to a possible role of these drugs in the prevention or regression of neoplastic processes. Amongst the proven anticancer effects of statins, some of them stand out such as: inhibition of cell proliferation, promotion of apoptosis, inhibition of angiogenesis and metastasis prevention. Thus, this study sough to elucidate simvastatin and pravastatin effects on progenitor cells and NSCs, and on some signaling pathways in breast carcinogenesis model (based on induction 7,12 dimethylbenz (a) anthracene [DMBA]) in female Sprague-Dawley rats. After a 14 days treatment with the statins, the rats' breasts were examined to verify immunostaining of progenitor cells and CSCs (CD133, CD24, CD44 and EpCAM), biological variables (tumor volume, mitosis, proliferation index) in addition to protein analysis of Akt and Src. The highest dose of the tested simvastatin (40mg/kg) decreased the number of tumors developed, volume and tumor growth as well as the cell proliferation index. There was no change in the percentage of necrosis to treatment with statins. Furthermore, simvastatin decreased the levels of Akt phosphorylation and increased PTEN levels, without significant differences in Src levels. Simvastatin was also able to reduce the number of CD133, CD24 and CD44 positive cells. For the doses tested, there was no difference on the analyzed parameters in the treatment with pravastatin. As a conclusion, in this model, chronic treatment with simvastatin showed cytostatic effects, regulatory actions towards Akt, as well as the control of CSCs and progenitor cells in the in vivo model of mammary carcinoma / Doutorado / Farmacologia / Doutor em Farmacologia

O Brasil, que conta hoje com 7% de idosos, será a sexta maior população de idosos do Planeta em 2020. A doença cardiovascular aumenta significativamente com a idade e evidências indicam a importância do endotélio e da hipercolesterolemia na aterosclerose. Contudo, não está bem estabelecido, que o tratamento da dislipidemia em idosos melhora a função vasomotora. Objetivos: Analisar a resposta vasomotora e parâmetros metabólicos em idosos com hipercolesterolemia ao tratamento com estatina. Delineamento: Ensaio Clínico Randomizado Duplo Cego. Casuística e Métodos: 72 idosos com idade 65 anos [75 ± 7,7 (65 a 92)], 64% mulheres, excluídos diabéticos, hipertensos graves, obesos, corticoterapia, reposição hormonal, doença em estágio terminal; todos realizaram avaliação clínica, laboratorial (glicose, lípides, TGO, TGP, CPK, Proteina C reativa ultra-sensível (PCR)) e função vasomotora por ultra-sonografia de alta resolução da artéria braquial, com equipamento ATL HDI 5000 e transdutor multifreqüencial de 7 a 10 MHz, segundo Diretrizes. Utilizou-se hiperemia reativa para resposta vasomotora dependente do endotélio (DMF) e nitrato sub-lingual para a independente do endotélio (DNT). Destes, 47 pacientes apresentaram LDL 130 mg/dL e foram randomicamente designados para grupo placebo (GP) e grupo tratamento (GT); todos realizaram avaliação antes e depois da intervenção (placebo ou atorvastatina 20mg oral por 30 dias). Para a análise estatística, utilizaram-se testes paramétricos e não-paramétricos, valores de p 0,05 foram considerados estatisticamente significantes. Resultados: Os grupos foram semelhantes em suas características basais; a DMF não apresentou diferença estatisticamente significante entre o GP e GT após intervenção [6,2 ± 6,2 (-4,8 23,7) e mediana = 6,4 vs. 5,0±5,6 (-5,1 18,9) e mediana= 3,5 p = 0,551], o mesmo ocorrendo para a DNT [7,1 ± 4,7 (-1,3 21,2) e mediana = 6,8 vs. 8,6±5,0 (1,4 21,6) e mediana = 6,8 p = 0,373). A análise estratificada sugere potencial modificador de efeito das co-variáveis estratificadas sexos, estratos etários (<80 e 80 anos) e níveis de LDL (< 160mg/dL e 160mg/dL) em relação à associação atorvastatina e função vasomotora. A PCR diminuiu 64% no grupo estatina (mediana no GP = 0,3 mg/dL vs. GT = 0,1 mg/dL p = 0,014). A redução do colesterol total foi de 27%, LDL = 41%, VLDL = 20% e triglicérides = 34%; o HDL elevou 12% no GT. A maior elevação enzimática não ultrapassou 3 vezes o limite e não ocorreram mialgias. Conclusões: Apesar da diminuição dos lípides, não houve modificações significativas da função vasomotora, o que sugere alterações intrínsecas do vaso associado ao envelhecimento. Os níveis de PCR reduziram significativamente, sugerindo um provável efeito antiinflamatório / The Brazil, where old people make up 7% of the population, will have the sixth greatest population of elderly people in the world. Cardiovascular disease increases significantly as a person grows older and evidences show the importance of the endothelium and hypercholesterolemia in arteriosclerosis. However, it has not been established yet if treating dyslipidemia in the elderly will improve this function. Objectives: To evaluate vasomotor response in elders suffering from hypercholesterolemia to the treatment with statin and to evaluate their lipids, c-Reactive Protein (CRP). Design: Double-blind randomized clinical trial. Methods: Seventy-two elderly people aged >65 years old [75 ± 7.7 (65 to 92)], 65% of women, excluding those suffering from diabetes, serious hypertension, obesity, steroid use, hormone replacement, end-stage disease; all of them underwent clinical evaluation, laboratorial evaluation (glucose, lipids, liver enzymes, CPK, high sensitivity CRP (CRP). Vasomotor response was evaluated by means of ATL HDI 5000 high resolution ultrasound scan of the brachial artery and multifrequency transduction of 7 to 10 MHz. Reactive hyperemia was used for endothelium-dependent flow response (DMF) and sublingual nitrate for the endothelium-independent dilation (DNT). Out of those patients, 47 presented LDL 130 mg/dl and were ramdonly divided into placebo (n = 23) and treatment group (n = 24); all of them had underwent evaluation before and after intervention (placebo or atorvastatin 20 mg, orally, during 30 days); for statistic analysis parametric and non-parametric tests were used and p 0.05 values were considered statistically significant. Results: The groups were similar in their basal characteristics; DMF did not present any statistically significant difference between placebo and treatment group after intervention [6.2 ± 6.2 (-4,8 - 23.7) and median = 6.4 vs. 5.0 ± 5.6 (-5.1 - 18.9) and median = 3.5 p = 0.551], the same occurred to DNT [7.1 ± 4,7 (-1.3 - 21.2) and median = 6.8 vs. 8.6 ± 5.0 (1.4 - 21.6) and median = 6.8 p = 0.373). Stratified analysis suggests effect modifying factor of the gender, age strata (< 80 and 80 years of age) and LDL (< 160 mg/dL e 160 mg/dL). CRP diminished 64% in the treatment group (median in the placebo group = 0.3 mg/dL vs. treatment group = 0.1 mg/dL p = 0.014). The reduction of total cholesterol was 27%, LDL = 41%, VLDL = 20% and triglycerides = 34%; HDL raised 12% in the treatmente group in relation to the placebo group. The highest enzymatic elevation did not surpass 3 times the limit and there were no myalgia. Conclusions: In spite of a significant decrease in lipids, there were no significant changes in vasomotion response with statin therapy, suggesting intrinsic alterations of the vessel associated with ageing. However, CRP reduction suggested an anti-inflammatory effect

No description available.

Introdução: A associação de uma estatina com ezetimiba é tão eficaz quanto altas doses da mesma estatina na redução do LDL-colesterol. Os efeitos que não dependem dessa redução são chamados de pleiotrópicos, entre os quais podemos citar: melhora da função endotelial, efeitos anti-oxidantes, efeitos anti- inflamatórios, entre outros. Objetivo: comparar a ação de dois esquemas de tratamento que obtêm reduções equivalentes de LDL-colesterol (sinvastatina 80 mg ao dia e associação sinvastatina 10mg/ezetimiba 10 mg ao dia), sobre os efeitos pleiotrópicos: inflamação, função endotelial e oxidação da LDL. Métodos: estudamos 23 pacientes randomizados e na forma de cross-over 2x2. A inflamação foi mensurada através da PCR-us, a função endotelial por meio de ultra-sonografia e a oxidação de LDL pelas dosagens de LDL eletronegativa (LDL-) e do anticorpo anti-LDL-. Resultados: A redução do LDL-colesterol foi similar nos dois grupos (45,27% no grupo sinvastatina/ezetimiba (p<0,001) e 49,05% no grupo sinvastatina (p<0,001), sem diferença entre os tratamentos (p=0,968)). Os dois grupos apresentaram melhora da função endotelial (3,61% no grupo sinvastatina/ezetimiba (p=0,003) e 5,08% no grupo sinvastatina (p<0,001), não houve diferença entre os tratamentos (p=0,291)). Houve melhora nos níveis da PCR-us (redução de -22,8% no grupo sinvastatina/ezetimiba (p=0,004) e de 29,69% no grupo sinvastatina (p=0,01), sem diferenças entre os tratamentos (p=0,380)). Não houve redução significativa da LDL-. Ocorreu aumento na concentração do anticorpo anti-LDL eletronegativa apenas no grupo sinvastatina (p=0,045). Conclusões: as duas formas de tratamento são eficazes na melhora da função endotelial e dos níveis de PCR-us. Somente com o uso da sinvastatina em alta dose houve aumento nos níveis de anticorpos anti-LDL-. / Introduction: The co-administration of a statin with ezetimibe is as effective as high doses of the same statin in the reduction of the LDL-cholesterol. The effects which don´t depend of this reduction are called pleiotropic effects, some among them can be cited: endothelial function improvement, antioxidative and anti-inflammatory effects. Objective: compare the effectiveness of these two different treatments that obtain equivalent reductions of LDLcholesterol (simvastatin 80 mg once a day and co-administration of simvastatin 10 mg once a day and ezetimibe 10 mg once a day), about pleiotropic effects: inflammation, endothelial function and LDL oxidation. Methods: we have studied 23 randomized patients in a 2x2 cross-over study. Inflammation was measured by high-sensitive C reactive protein, endothelial function by echocardiography and LDL oxidation by electronegative LDL and electronegative anti-LDL antibodies levels. Results: the LDL-cholesterol was similar between the two groups (45,27% reduction in the simvastatin/ezetimibe group (p<0,001) and 49,05% reduction in the simvastatin group (p<0,001); no difference between treatments was found (p=0,968). The two groups had improvement in endothelial function (3,61% in the simvastatin/ezetimibe group (p=0,003) and 5,08% in the simvastatin group (p<0,001)), no differences was found between the two groups (p=0,291). High-sensitive C reactive protein had a 22,8% reduction in the simvastatin/ezetimiba group (p=0,004) and 29,69% reduction in the simvastatin group (p=0,01), with no significative difference in any of the two treatments (p=0,380). There was no significative difference in LDL- levels. The anti-LDL- antibodies concentration was increased only in the simvastatin group (p=0,045). Conclusion: the two forms of treatments presented some similar pleiotropic effects - improvement in endothelial function and decreased hs-CRP levels. Only with a high simvastatim dose the anti-LDL- antibodies concentration was increased.

HMG-CoA-Reduktasehemmer, sogenannte Statine, und regelmäßige körperliche Aktivität sind mit vermindertem Auftreten zerebrovaskulärer Ereignisse und Zunahme der endothelialen Stickstoffmonoxidsynthase (eNOS) assoziiert. Die Erhöhung der eNOS-mRNA ist mit verbessertem zerebralen Blutfluß und Neuroprotektion bei einer zerebralen Ischämie verbunden. Vor dem Hintergrund, daß Thrombosen und Thrombembolien die häufigste Ursache zerebro- und kardiovaskulärer Ereignisse darstellen, sind NO-vermittelte antithrombotische Effekte jedoch kaum untersucht. Ebenso wenig ist über mögliche Absetzeffekte nach Beendigung einer Statintherapie bekannt. Daher untersuchten wir, ob die Statine Atorva- und Rosuvastatin eNOS-abhängig zu Neuroprotektion führen und verglichen die Effekte mit einem zweiten eNOS-regulierenden Mechanismus: der regelmäßigen körperlichen Aktivität. Dazu quantifizierten wir nach entsprechender Vorbehandlung eNOS auf mRNA- und Proteinebene aus Aorten, Hirngewebe sowie Thrombozyten und bestimmten die Läsionsvolumina im experimentellen Schlaganfallmodell. Außerdem untersuchten wir nach Statingabe Thrombozytenfunktionsparameter sowie Blutungszeit und Thrombusformation in vivo. Zwei bzw. vier Tage nach Absetzen der Statinbehandlung wiederholten wir die eNOS-Messungen, Schlaganfallexperimente und Gerinnungsanalysen. Wir fanden nach Statinvorbehandlung cholesterinunabhängig eine Zunahme der eNOS, was mit Neuroprotektion im Schlaganfallmodell und verminderter Gerinnungsaktivität verbunden war. Nach Absetzen der Behandlung kam es jedoch zu einer drastischen Abnahme der eNOS, was mit deutlichem Anstieg der Thrombozytenmarker im Plasma und schnellem Verlust der beobachteten positiven Effekte auf Läsionsgröße und Gerinnungssystem einherging. Regelmäßige körperliche Aktivität führt ebenfalls eNOS-abhängig zu verbessertem zerebralen Blutfluß und kleineren Läsionsvolumina bei zerebraler Ischämie. Diese Ergebnisse sind mit den Daten nach Statingabe vergleichbar. Wir demonstrieren einen Klasseneffekt der Statine für eNOS-vermittelte Neuroprotektion im zerebralen Ischämiemodell. Durch die zusätzliche gerinnungshemmende Wirkung könnte diese Wirkstoffklasse neue Ansätze zur prophylaktischen Schlaganfallbehandlung unabhängig vom Cholesterinspiegel eröffnen. Ein Absetzen der Statinbehandlung kann jedoch zu einer Zunahme der Schlaganfallgröße führen und sollte möglicherweise bei Risikopatienten vermieden werden. Regelmäßiges körperliches Training führt zu vergleichbarer Erhöhung der eNOS sowie Neuroprotektion und bietet damit eine sinnvolle Verknüpfung aus prophylaktischer Schlaganfallbehandlung und Rehabilitation. / HMG-CoA-reductase inhibitors, so called statins and regular physical activity are associated with less cerebrovascular events and increase of endothelial nitric oxide synthase (eNOS). Raise of eNOS-mRNA results in cerebral blood flow (CBF) augmentation which refers neuroprotection after ischemic stroke. It is known that thromboses cause the most cerebrovascular events, but nitric oxide (NO) dependent antithrombotic effects are poor examined. In addition there are little information about effects after withdrawal of statin treatment. That is why we investigated Atorva- and Rosuvastatin regarding eNOS dependent neuroprotection and compared the effects with regular physical activity, the second eNOS enhancing mechanism. Therefore after corresponding pretreatment we quantified eNOS-mRNA and protein from aortas, brain tissue and thrombocytes and determined lesion volume after experimental middle cerebral artery occlusion (MCAo). Furthermore after statin treatment we measured marker of thrombocyte activation, as well as bleeding time and thrombus formation in vivo. Two and four days after withdrawal of statin treatment we repeated eNOS measurements, neuroprotection studies and coagulation analyses. We found eNOS upregulation independent from serum cholesterol level after statin pretreatment and this was associated with neuroprotection after ischemic stroke and decreased platelet activation. But after withdrawal of statin treatment eNOS expression was downregulated, which went along with clear upregulation of platelet activation and a rapid loss of the observed positive effects on lesion volume and hemostasis. Regular physical activity leads to an increase of eNOS, which we could correlate with CBF augmentation and improved outcome after MCAo. These results were comparable to the data after statin treatment. We demonstrate a class effect of statins for eNOS-dependent neuroprotection in our ischemia modell. Because of the additional antithrombotic effects statins may present a new approach to prophylactic stroke treatment independent from cholesterol level. Withdrawal of statin treatment may refer increased cerebral lesion volume and should be avoided in patients with risk for cerebrovascular events. Regular physical activity results in comparable eNOS dependent neuroprotection and offers a useful combination between prophylactic stroke treatment and rehabilitation.

Introdução: As avaliações econômicas completas do tipo custo-utilidade, suportadas por dados de efetividade do mundo real, permitem uma perspectiva diferenciada da avaliação de tecnologia em saúde. Objetivo: Realizar uma análise de custo-utilidade do uso de estatinas para a prevenção secundária de eventos cardiovasculares em portadores de doença cardiovascular isquêmica, e avaliar a variabilidade da efetividade e da razão de custo-efetividade incremental com diferentes classificações de usuários de estatina (incidentes e prevalentes). Método: Um modelo de microssimulação de Markov com 5 estados, ciclos anuais e horizonte temporal de 20 anos, com taxas de desconto de 5% foi desenvolvido. As probabilidades de transição para mortalidade por todas as causas foram extraídas após pareamento por escore de propensão dos dados e tratamento de dados ausentes de uma base secundária de registro assistencial com linkage determinístico com a base de mortalidade do Ministério da Saúde. As probabilidades dos desfechos não-fatais foram obtidas na literatura. As medidas de efetividade (QALY) foram calculadas com dados publicados dos domínios do SF-36 de um estudo realizado com a população do mesmo hospital, no mesmo período e com as mesmas condições clínicas e transformados em medidas de utilidade por modelo validado. Somente custos diretos na perspectiva do reembolso do SUS foram considerados. Resultados: 3.150 pacientes foram pareados após o escore de propensão, 1.050 não usuários de estatina, 1.050 usuários de estatinas classificados como prevalentes e 1.050 usuários de estatinas classificados como incidentes, com diagnóstico de doença cardiovascular isquêmica prévia, com seguimento médio de 5,1 anos. A efetividade das estatinas quando considerados todos os usuários em relação aos não usuários resultou em um HR para mortalidade de 0,992 (IC 95% 0,85; 0,96) e de 0,90 (IC 95% 0,85; 0,96) para os usuários incidentes. A RCEI comparando todos os usuários de estatinas versus não usuários foi de R$5.846,10/QALY e de R$7.275,61/QALY para os usuários incidentes. Conclusão: As estatinas diminuíram a mortalidade por todas as causas, e a análise incluindo usuários prevalentes diminui o tamanho do efeito. O tratamento possui custo-efetividade favorável dentro do limiar de disponibilidade a pagar definido, sendo modificado pela forma de extração do dado de efetividade / Background: The complete economic evaluations, such as cost-utility analysis, supported by real world data of effectiveness lead to a more realistic perspective of a health technology assessment. Objective: Perform a cost-utility analysis of statins for secondary prevention in ischemic cardiovascular disease patients based on effectiveness from real world data and evaluate the variability of effectiveness and incremental cost-effectiveness ratio (ICER) considering prevalent and incident users. Methods: A Markov microssimulation model with five states, annual cycle and time horizon (TH) of 20 years, with discount rate of 5% was developed. Transition probabilities for all cause mortality was derived from a secondary database of a teaching hospital after record linkage with national registry of mortality database and an analysis of propensity score matching and multiple imputation analysis for missing data. Non-fatal endpoints were derived from a published meta-analysis. Utility measures was calculated with a validated model to derive values from published domains of SF-36 QoL questionnaire, domains was measured for a published RCT in the same teaching hospital, over the same period, with similar age and diagnostic characteristics. Only direct costs were analyzed from the Brazilian public health reimbursement perspective. Results: 3150 patients were matched, 1050 non-statins users (CG), 1050 prevalent statins users (PSU) and 1050 incident statins users (ISU) with previous cardiovascular disease, with mean follow-up of 5,1 years. Treatment effects on the treatment group considering all statins users for all cause mortality had a hazard ratio of HR:0,992 (IC 95% 0,85 - 0,96) and HR: 0,90 (IC 95% 0,85 - 0,96) only for ISU. The ICER comparing all users with non-users was R$5.846,10 per QALY and for ISU was R$7.275,61 per QALY. Conclusion: Real world evidence demonstrated that statins are an effective treatment to reduce all cause mortality in secondary prevention and are a cost-effective strategy considering the willingness to pay established, but the prevalent users resulted in less effectiveness of the drug when included in the analysis and influenced the ICER

Clinical evidence suggested patients with lower plasma C-reactive protein (CRP) levels after statin therapy could have better clinical outcome. The last part of the study was to measure on-treatment high sensitivity CRP (hsCRP) levels among 229 Chinese patients with hyperlipidaemia undergoing treatment with simvastatin 40 mg daily. The patients were genotyped for 15 SNPs or haplotypes in 11 candidate genes that would have significant allele frequency among Chinese patients and may be linked to statin efficacy or hsCRP levels. The analysis suggested BMI is the largest single contributing factor of 15.0% of the variation in hsCRP levels, followed by plasma triglycerides levels contributing 4.7% and male gender 1.6% (all P&lt;0.05). However comparisons of hsCRP levels among genotype groups did not reveal any significant findings, with or without adjustment with covariate genotypic or phenotypic factors. To further categorize individuals as high or medium risk, we set a threshold hsCRP level of 1 mg/L as the benchmark for evaluation. The CRPc.3872G>A SNP was related to lower risk compared to the homozygous wild-type genotype (adjusted odds ratio AOR = 0.289; P = 0.014) after adjusting for phenotypic factors of age, gender, smoking status, BMI, waist circumference, hip circumference, plasma lipid profiles, co-existing disease and co-medications. Another marginal finding included the HNF1A c.79A>C SNP (AOR = 0.575; P = 0.118). / Polymorphisms in the drug transporters are likely to be more important with hydrophilic statins such as pitavastatin, which undergoes transporter mediated distribution. The SLCO1B1 c.388A>G polymorphism in the gene encoding the uptake transporter organic anion transporting polypeptide (OATP1B1) is common in Chinese and the variant was associated with increases of 63--68% in maximum plasma concentration and 44--47% in systemic exposure of both the lactone and acid compared to wild-type subjects (P&lt;0.05). Co-administration of pitavastatin with grapefruit juice (GFJ) resulted in a small increase of the area under the plasma concentration time curve (AVC) by 15--16% for both the acid and lactone (P&lt;0.05). However, there was no significant effect on the drug-food interaction in relation to relevant SNPs in the enzymes and transporters examined. / The SNPs examined included those in the genes for the enzymes and transporters involved in the metabolic pathway or the distribution of simvastatin. Cytochrome P450 (CYP) enzymes are involved in hepatic and intestinal metabolism of several statins and simvastatin is known to undergo extensive metabolism via the CYP3A4/3A5 pathway. The common candidate SNPs in the CYP3A4/3A5 enzymes found in Chinese populations include CYP3A4*1G, CYP3AP1*3 and CYP3A5*3 , which are associated with altered enzyme expression and activity. However, no statistically significant relationship was found between these SNPs and a potential phenotypic marker of enzyme activity, the urinary ratio of 6beta-hydroxy-cortisol/cortisol (6beta-OHC/C) concentrations. The analysis of lipid lowering responses in relation to individual SNPs or combinations from gene-gene interactions also revealed no statistically significant findings. In the subgroup of patients with familial hypercholesterolaemia, the CYP3A4*1G, CYP3AP1*3 and CYP3A5*3 polymorphisms appeared to have a small effect on the changes in LDL-C and total cholesterol with the subjects with the CYP3A5*3 and CYP3AP1*3 variants showing less reduction and those with the CYP3A4*1G variant showing more reduction than subjects with the wild-type genotype with a tendency for a gene-dose effect. It is difficult to interpret these findings and the significance may be related to multiple testing. / The statins, or 3-hydroxymethyl-3-glutaryl coenzyme A (HMG-CoA) reductase inhibitors, act on the rate limiting step in endogenous cholesterol synthesis. Their primary action results in reduction of plasma low-density lipoprotein cholesterol (LDL-C) levels and this is thought to be the major mechanism by which they reduce cardiovascular events. There are considerable differences between subjects in both the plasma levels of the statins and in their effects on LDL-C and other lipid parameters and some of this variation appears to be related to genetic differences in the pathways of drug metabolism and distribution and in the pathways involved in lipid metabolism. / The variation in response may be related to variations in systemic or hepatic exposure to the drug, which in turn will be related to the pharmacokinetics. This is also likely to play a role in the adverse effects of myopathy and therapeutic tolerance. In a pharmacokinetic study in healthy male Chinese subjects, the common polymorphism of CYP2D6*10 was analyzed in relation to the pharmacokinetics of lovastatin and simvastatin. There was a tendency for reduced clearance of simvastatin lactone by 30% (P>0.05) in subjects with the CYP2D6*10/*10 genotype. With lovastatin, there were similar findings with 38.5--84.9% decrease in clearance which appeared to be related to enzyme activity according to genotype, with *5 carriers showing a greater decline in clearance than *10 carriers (P&lt;0.05). / These results provide some insights into the pharmacokinetics and pharmacodynamics of statins and the pharamacogenetic relationships to candidate SNPs. Future research in this field should help to facilitate safer and more effective treatment with these commonly used medications, resulting in personalized therapy and optimal clinical benefits for patients with cardiovascular disease. / This thesis describes a study of 270 patients recruited from the outpatient clinics at the Prince of Wales Hospital who were treated with simvastatin 40 mg daily for at least 4 weeks. Their mean (+/-SD) LDL-C baseline level was 5.38+/-1.68 mmol/L and the reduction in LDL-C after simvastatin treatment was 2.81+/-0.99 mmol/L or -47.1+/-12.5%. / Mak, Wah Lun Valiant. / Adviser: Brian Tomlinson. / Source: Dissertation Abstracts International, Volume: 73-06, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2011. / Includes bibliographical references (leaves 253-289). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

A indicação de biópsia para o diagnóstico precoce do câncer prostático baseia-se na dosagem sérica do PSA e nos achados do toque retal. O PSA é uma kalecreina estando seus genes reguladores ligados aos andrógenos. Drogas que afetam o metabolismo dos andrógenos podem afetar a produção de PSA. A finasterida é uma droga sintética que inibe a conversão de testosterona em DHT pela enzima 5 AR. O uso da finasterida na dose de 5mg/dia para o tratamento da HPB causa redução do volume prostático de 20 a 30% e diminuição dos valores dos PSA em aproximadamente 50% do seu valor inicial após 6 meses. O uso da finasterida na dose de 1mg/dia para o tratamento da AAM foi aprovado pelo FDA em 1997. Um estudo realizado em 2007 avaliou a alteração do nível do PSA em homens com mais de 40 anos fazendo uso de finasterida 1mg/dia para tratamento da AAM. Os resultados revelaram redução dos valores do PSA semelhante à verificada nos pacientes portadores de HPB. Não existem estudos prospectivos sobre o tema incluindo pacientes mais jovens. O objetivo do nosso trabalho foi verificar as alterações da dosagem do PSA, da testosterona sérica total e do volume prostático em indivíduos com menos de 40 anos de idade com em uso de finasterida 1mg/dia. Selecionamos 52 pacientes que após avaliação inicial preenchiam os critérios de inclusão. Foram dosados os níveis séricos do PSA e da testosterona, e mensurado o volume prostático através da ultrassonografia transabdominal, no início do estudo (T0) e um ano após o uso da finasterida (T2). No intervalo, 6 meses após o início da droga, foi solicitada apenas nova dosagem de PSA (T1). O valor médio na avaliação inicial (T0) da dosagem do PSA, da testosterona total plasmática e do volume prostático mensurado pela ultrassonografia transabdominal foi de 0,398 ng/ml (0,14- 0,78); 735,77 ng/dl (548-927) e 21,35 ml (15-31ml) respectivamente. Foi observada uma redução do valor médio do PSA de 9,21% após 6 meses do uso da droga (p=0,001). Após 12 meses do uso da finasterida verificamos uma redução de 10,51% do valor do PSA em relação à dosagem inicial (p<0,001) e uma diminuição do valor médio do volume prostático 21,37 ml para 20,03 ml (p<0,001). Não foi detectada alterações nos níveis de testosterona. Diferentemente de estudos anteriores em que, em homens fazendo uso de finasterida 1mg/dia, houve redução de 40% e 50% dos valores do PSA nas faixas etárias de 40 a 49 anos e 50 a 59 anos, nosso trabalho revelou reduções inferiores. Nosso estudo traz importantes questionamentos em relação a como deve ser feita a correção dos valores do PSA nos pacientes que começaram a utilizar finansterida 1mg antes dos 40 anos de idade e que se apresentam para avaliação prostática. Outro ponto de interesse é se a hiperplasia do componente epitelial observada durante envelhecimento masculino poderia ser inibida pelo uso da finasterida desde a juventude, pois sabemos que indivíduos portadores de deficiência congênita de 5AR não apresentam alopecia e nem tão pouco desenvolvem HPB. / The indication of biopsy for early diagnosis of prostate cancer is based on serum PSA levels and digital rectal examination findings. PSA is a kalecreine and its regulatory genes are modulating by androgens. Drugs affecting the androgens metabolism can affect the production of PSA. Finasteride is a synthetic drug, which inhibits the conversion of testosterone to DHT by the enzyme 5 AR. There are two subtypes of 5AR enzymes, Type 1 that predominates in non-prostatic tissues such as liver and skin, and Type 2, which predominates in the prostate and scalp but is also expressed in other tissues. The use of Finasteride 5mg / day for the treatment of BPH is well known. After six months we observe a decrease in prostate volume by 20 to 30% and also a decrease in total serum PSA concentration of approximately 50%. The use of Finasteride at 1mg / day for the treatment of MAA has been approved by the FDA in 1997. In 2007 a study evaluated the change in total serum PSA concentration in men over 40 years taking Finasteride 1 mg / day to treat MAA. The results showed a reduction in PSA values similar to that observed in patients treated of BPH. There are no prospective studies including younger patients. The aim of our study was to assess the changes in total serum PSA concentration, total serum testosterone concentration and prostate volume in patients younger than 40 years of age in use of Finasteride 1mg / day for MAA. We prospectively selected 52 patients with MAA and indication for treatment with Finasteride who met the inclusion criteria. Serum levels of total PSA and total testosterone and prostate volume, measured by transabdominal ultrasound, were obtained at baseline (T0) and one year after started the drug (T2). In the interval, 6 months after started drug, only serum total PSA concentration was measured (T1). The median value at baseline (T0), for total PSA, total testosterone and prostate volume was 0.398 ng / ml (0.14 to 0.78); 735.77 ng / dl (548-927) and 21.35 ml (15-31ml) respectively. A reduction of 9.21% on total PSA concentration was detected 6 months after started the drug (p = 0.001). After 12 months we observed a 10.51% decrease in total serum PSA concentration, when compared with the baseline value, (p <0.001). A reduction in the median value of prostate volume from 21.37 ml to 20.03 ml (p < 0.001) was also detected at the same period. There were no detectable changes in testosterone levels. They reported a decrease of 40% and 50% in total PSA concentration for groups between 40-49 and 50-59 of age respectively. In our study we observed lower reductions. Our finding may be explained by the fact that the epithelial component of the prostate gland has not yet started the hyperplasia process, so the conversion of testosterone into DHT by blocking 5AR by Finasteride would cause less impact. The fact that the values of plasma testosterone not have changed is not surprising since Finasteride is not considered an anti androgenic drug, it means that the synthesis of the testosterone is not affected by its use. Our study highlights important questions about how the adjustment of PSA values should be done in patients who began taking Finasteride 1mg / day before 40 years of age and present for prostate evaluation. Another point of interest is whether the hyperplasia of the epithelial component, observed during the aging process, could be inhibited by the use of Finasteride. This hypothesis can be corroborate by the fact that individuals with congenital deficiency of 5AR do not present alopecia or develop BPH. In conclusion the use of Finasteride 1mg /day reduces the total serum PSA value by 10,51% after one year. Prostate volume is also reduced by 6,3% in the same period.

Les données épidémiologiques montrent une association inverse entre les taux de HDL-cholestérol (HDL-C) et le risque d'évènements cardiovasculaires. Des traitements ayant montré une augmentation significative du HDL-C, comme les inhibiteurs de la protéine de transfert des esters de cholestérol, devraient donc permettre de réduire le risque cardio-vasculaire. En utilisant différentes techniques de modélisation, nous avons tenté de quantifier l'efficacité attendue sur les événements cardiovasculaires de l'un d'entre eux, le dalcétrapib, ne disposant que de données pharmacocinétiques et pharmacodynamiques. Tout d’abord, afin d'établir la relation pharmacocinétique / pharmacodynamique entre les concentrations et la modification de HDL-C, nous avons analysé les données individuelles des patients dyslipidémiques par une approche de population. Une hausse moyenne de HDL-C de 26.4 % par rapport au placebo était alors anticipée. Nous avons ensuite tenté de corréler l'effet observé sur l'HDL-C et l'effet clinique à partir de données d'autres études par méta-régression des essais évaluant l'effet des principaux hypolipémiants en prévention secondaire. Cette modélisation n'a pas permis de montrer de corrélation entre le changement de l’HDL-C (P5 P95 :-3.0 et 36 %) et la réduction du risque cardiovasculaire. Une analyse de sensibilité par type de traitement suggère qu'une même hausse de HDL-C entre deux classes thérapeutiques pourrait se traduire par un effet clinique dissemblable, indiquant que HDL-C ne peut pas être utilisé comme critère intermédiaire puisqu'il ne serait pas un prédicteur indépendant du risque cardiovasculaire / Epidemiological data demonstrate an inverse correlation between HDL-cholesterol (HDL-C) levels and cardiovascular risk. Therefore, drugs as cholesteryl ester transfer protein (CETP) inhibitors that lead to a significant HDL-C increase are believed to reduce the occurrence of coronary events. We aimed to evaluate the clinical efficacy of one CETP inhibitor, dalcetrapib, by using various modeling techniques while only pharmacokinetic (PK) and pharmacodynamlc (PD) data were available. First, we analyzed individual data from dyslipidemic patients using a population approach in order to establish the PK/PD relationship between dalcetrapib concentrations and HDL-C change. The results show that an average raise of 26.4 % is expected in comparison to placebo with the 5th (P5) and 95th (P95) percentile of the mean average at 20.7 % and 31.9 % respectively. The increase in HDL-C is explained by a delayed catabolism following the transfer inhibition of cholesterol ester from HDL to Apo-B rich lipoproteins. We endeavored then to correlate HDL-C increase to coronary events by using a meta-regression analysis on randomized trials that evaluated the clinical efficacy of main dyslipidemic drugs on coronary events in secondary prevention. The modeling did not show a statistical association between HDL-C change (P5-P95:-3.0 and 36 %) and coronary risk reduction. A sensitivity analysis by drug class suggests that the same HDL-C increase resulting from different mechanisms of action may not impact the cardiovascular risk in the same way. This would indicate that HDL-C could not be used as a risk marker since it might not be an independent predictor of cardiovascular risk