Submitted by Cláudia Bueno (claudiamoura18@gmail.com) on 2016-02-04T10:23:14Z No. of bitstreams: 2 Tese - Flavio Fernandes Veloso Borges - 2015.pdf: 2110689 bytes, checksum: 595ebd21ecf4f13568b0e3179e801f99 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2016-02-04T10:43:08Z (GMT) No. of bitstreams: 2 Tese - Flavio Fernandes Veloso Borges - 2015.pdf: 2110689 bytes, checksum: 595ebd21ecf4f13568b0e3179e801f99 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2016-02-04T10:43:08Z (GMT). No. of bitstreams: 2 Tese - Flavio Fernandes Veloso Borges - 2015.pdf: 2110689 bytes, checksum: 595ebd21ecf4f13568b0e3179e801f99 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2015-03-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Silymarin (SM) is a standardized extract from the seeds and leaves of milk thistle Silybum marianum (L.) Gaertn. It is composed mainly of flavonolignans, with silibinin (SB) being its principal active constituent. Known mainly as antioxidant and hepatoprotector, SM and SB were found to be clinically effective in the treatment of a variety of liver disorders, including acute and chronic viral hepatitis, toxin and drug-induced hepatitis and cirrhosis. Due to the wide biological activities presented by SM and SB, the present study aimed to evaluate their antimutagenic activities using the Ames mutagenicity test in Salmonella typhimurium, their antigenotoxic activities using the mouse bone marrow micronucleous test and the alkaline comet assay, and to assess their effect on the gene expression pattern of some genes associated with the process of carcinogenesis and chemoprevention. To assess antimutagenicity, bacterial suspensions of Salmonella typhimurium (TA98 and TA100 strains) were treated with different concentrations of SM or SB simultaneously with the appropriate positive controls for each strain. To assess antigenotoxicity, Swiss mice were orally treated with different concentrations of SM or SB simultaneously with a single intraperitoneal dose of mitomycin C (MMC) for the micronucleus test, and human blood lymphocytes were cotreated with SM or SB and methyl methanesulfonate (MMS) for the alcaline comet assay. To investigate the role of SM and SB in modulating gene expression, we conducted microarray analysis. The results showed that SM was not significantly effective in reducing the number of frameshift mutations in strain TA98, while SB demonstrated significant protection at higher doses (p < 0.05). Regarding strain TA 100, SM and SB significantly decreased mutagenicity (point mutations) (p < 0.05). The results of the antigenotoxic evaluation demonstrated that SM and SB significantly reduced the frequency of micronucleated polychromatic erythrocytes (MNPCE) (p < 0.05). The results also indicated that SM and SB significantly attenuated MMC induced cytotoxicity (p < 0.05). In the comet assay, SM and SB significantly reduced the genotoxicity of MMS (p < 0.05), with a stronger antigenotoxic activity exerted by the extract complex (SM) than the one exerted by the isolated main active constituent (SB). The expression array analysis of five genes related to DNA damage, carcinogenesis and/or chemoprevention mechanisms demonstrated an up-regulation of PTEN and BCL2, down-regulation of BAX and ABL1 and no significant change in ETV6 expression levels.In conclusion, our results demonstrated that both SM and SB presented antimutagenic and antigenotoxic actions, as well as modulated the expression levels of genes analysed under the experimental conditions of this study. / A silimarina (SM) é um extrato padronizado obtido a partir das sementes e folhas de Silybum marianum (L.) Gaertn. SM é composta principalmente de flavonóides, sendo a silibinina (SB) seu principal componente ativo. Conhecidas principalmente como antioxidantes e hepatoprotetoras, SM e SB foram consideradas clinicamente eficazes no tratamento de uma variedade de doenças do fígado, incluindo hepatites virais agudas e crônicas, hepatites induzidas por toxinas e/ou drogas e cirrose. Assim, devido à ampla gama de atividades biológicas apresentadas pela SM e SB, o presente estudo teve como objetivo avaliar suas atividades antimutagênicas utilizando o teste de Ames em Salmonella typhimurium, suas atividades antigenotóxicas pelo teste do micronúcleo em medula óssea de camundongos e pelo teste do cometa em linfócitos humanos e avaliar seus efeitos nos perfis de expressão gênica de alguns genes associados ao processo de carcinogênese e quimioprevenção. Para a avaliação da antimutagenicidade, suspensões bacterianas de Salmonella typhimurium (cepas TA98 e TA100) foram co-tratadas com diferentes concentrações de SM ou SB e os controles positivos adequados para cada cepa. Para a avaliação de antigenotoxidade, camundongos Swiss foram tratados oralmente com diferentes concentrações de SM ou SB concomitantemente a uma única dose intraperitoneal de mitomicina C (MMC) para o teste do micronúcleo, e linfócitos humanos foram tratados simultaneamente com SM ou SB e metil-metanossulfonato (MMS) para o ensaio do Cometa. Os resultados mostraram que a SM não foi significativamente efetiva em reduzir o número de mutações com deslocamento de quadro de leitura na cepa TA 98, enquanto que a SB apresentou uma proteção significativa nas doses maiores (p < 0.05). Em relação à cepa TA100, SM e SB reduziram significativamente a mutagenicidade (mudanças de pares de bases) (p < 0.05). Na avaliação de antigenotoxidade, SM e SB reduziram significativamente a frequência de eritrócitos policromáticos micronucleados (EPCMN) (p<0,05). Os resultados também mostraram que a citotoxicidade causada pela MMC foi significativamente atenuada pela SM e SB (p<0,05). No ensaio do cometa, SM e SB reduziram significativamente a genotoxicidade provocada pelo MMS (p<0.05), com uma atividade antigenotóxica maior exercida pelo extrato complexo (SM) do que pelo principal componente ativo isolado (SB). A análise dos níveis de expressão de cinco genes relacionados ao dano no DNA, mecanismos de carcinogênese e/ou quimioprevenção demonstrou um aumento na expressão de PTEN e BCL2, diminuição na expressão de BAX e ABL1 e ausência de mudança significativa nos níveis de expressão do ETV6. Com base nesses resultados, conclui-se que a SM e a SB apresentaram ações antimutagênicas e antigenotóxicas, e também modularam os níveis de expressão dos genes analisados sob as condições experimentais deste estudo.

This thesis is aimed at studying the effect of the addition of milk thistle (Silybum marianum) seed cakes to the diet on metabolism, liver function and performance of laying hens at the beginning of laying. In the experiment, the results were compared between a group of laying hens, which were fed with the basic feed mixture without the addition of milk thistle seed cakes with the experimental group with the addition of milk thistle seed cakes in the feed mixture. It was situated 30 hens breed ISA Brown into the experiment, divided into two groups. The first group was experimental with the addition of milk thistle seed cakes (7 % in the mixture), and the second group was a control. The experiment evaluated egg laying, egg weight, food consumption and the animal weight. During the experiment, blood was twice collected from animals for biochemical examination focusing on the parameters of liver profile. The results of the experiments didn´t show any effects of feeding milk thistle seed cakes on the body weight of laying hens. By evaluating the performance parameters of the laying hens it showed a significantly lower egg laying, but also showed a lower consumption of feed (P < 0.05). At the same time, the data found higher weigh of eggs (P < 0.01) in this group, but the egg production was significantly lower than in the control group. During the examination of biochemical parameters of blood there was a significantly higher concentration of bilirubin, uric acid and cholesterol (P < 0.05) in the blood plasma of laying hens fed by the addition of milk thistle seed cakes. At the second sampling (approximately 2 months) the laying hens that were fed with milk thistle seed cakes have consistently higher concentration of cholesterol (P < 0.05) and lower activity of dehydrogenase, creatine kinase (P < 0.05) and uric acid (P < 0.01) in the blood. In the experiment there was no clear positive or negative effects of the milk thistle seed cakes on the performance parameters and biochemical profile of the blood plasma.

Heme oxygenase (HMOX) catalyzes first and rate-limiting step in heme degradation. By its action, carbon monoxide (CO), ferrous iron and biliverdin which is subsequently reduced to bilirubin are produced. Before discovery of HMOX reaction mechanism, CO was considered only a toxic waste product without any significant importance for human organism. Bilirubin, marker of liver dysfunction, has been also exposed to similar perception. But results from past decades show that HMOX and its metabolic products play an important role in number of physiological as well as defense against pathophysiological processes. The aim of this thesis was to clarify the role of HMOX and its metabolic products, presumably CO and bilirubin, in vivo and in vitro. We focused on the role of CO in a rat model of lipopolysaccharide-induced cholestasis. We were first to describe tissue distribution and pharmacokinetics of inhaled CO in this animal model and found out that CO inhalation is associated with anti-inflammatory and hepatoprotective effects. In a rat model of ethinylestradiol-induced cholestasis, we demonstrated the anticholestatic effect of HMOX. The induction of HMOX by its substrate heme increased the expression of liver transporters thereby increasing bile flow and simultaneously facilitated effective clearance of...

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Tyrimo objektas ir metodai: eksperimentinės augalinio vaistinio preparato Silymarinum – Aconitum 140 mg kietos kapsulės, kurių veiklioji medžiaga yra silimarinas. Kiekybinė jų sudėtis nustatyta ESC metodu. Darbo tikslas: pagrįsti augalinio vaistinio preparato kiekybinį stabilumą validuotu ESC metodu. Darbo uždaviniai: atlikti mokslinės literatūros šaltinių analizę įvertinant pasirinkto vaistinio augalinio preparato savybes bei aktyvių junginių nustatymo metodikas. Optimizuoti ir validuoti ESC metodiką kiekybiniam pasirinkto vaistinio augalinio preparato įvertinimui. Validuotu metodu atlikti eksperimentinių augalinio vaistinio preparato serijų stabilumo kiekybinę analizę. Apdoroti ir įvertinti augalinio vaistinio preparato stabilumo duomenis, remiantis mokslinių gairių nurodytais kriterijais ir gautais rezultatais pagrįsti preparato stabilumą. Išvados: atlikta mokslinės literatūros šaltinių analizė, pagrindžianti gydomąsias augalinio vaistinio preparato Silymarinum – Aconitum 140 mg kietomis kapsulėmis savybes, aktyvių junginių nustatymo metodikas bei taikomus metodo optimizavimo ir validacijos kriterijus. Optimizuotas ESC metodas eksperimentinio augalinio vaistinio preparato Silymarinum – Aconitum 140 mg kietomis kapsulėmis kiekybinei analizei. Validacijos eksperimentų metu pagrįstas metodikos glaudumas ir tiesiškumas. Atlikta kiekybinė eksperimentinio augalinio vaistinio preparato Silymarinum – Aconitum 140 mg kietomis kapsulėmis mėginių analizė. Preparato mėginiuose... [toliau žr. visą tekstą] / Object and methods: experimental herbal medicinal product Silymarinum – Aconitum 140 mg in hard capsules, as the main active component is silimarin. Assay analysis has been performed by HPLC method. Aim: to justify herbal medicinal product quantitative stability by validated HPLC method. Objective: to perform analysis of scientific literature and evaluate characteristics of chosen herbal medicinal product, and analysis methods of active components. Optimize and validate HPLC method for quantitative evaluation of chosen herbal medicinal product. Perform stability indicating quantitative analysis of experimental herbal medicinal product. Evaluate stability data of herbal medicinal product and justify its stability with reference to scientific guidelines and gained results. Results: performed scientific literature analysis justified therapeutic effect of herbal medicinal product Silymarinum – Aconitum 140 mg in hard capsules, evaluated analysis methods of active components and applicable criteria for method optimization and validation. HPLC method for quantitative analysis of experimental herbal medicinal product Silymarinum – Aconitum 140 mh in hard capsules was optimized. Precision and linearity of method was justified during experiments of validation. Assay analysis of herbal medicinal product Silymarinum – Aconitum 140 mg in hard capsules clarified, that amount of total silymarin varies from 136,966 mg to 146,573 mg. Stability testing clarified that herbal medicinal... [to full text]

Milk Thistle (Silybum marianum) has been one of the best known medici herb for a very long time. Seeds contain biologicky aktive substances taxifoiln, silychristin, silydianin, silybin A, silybin B, isosilybin A and isosilybin B, commonly known as silymarin complex. The aim of this thesis was the influence of elicitors, fertilization and growing technology on this product and its utilizaion. In the first part I focused on history, botanic attributes, agrotechnology, chemical composition and substance efficiency method of determinativ and also on pharmacological onpacts of effective substances. In the practical part I conducted twho small parcel experiments to reveal the impal of elicitors on effective substances. I used two elicitors NanoFYTSi? in 1ml/l concentrantion and N-FENOLMIX? in 0.5ml/l concentration. In the second experiment from my supervisor elicitor NanoFYT Si? in 1ml/l concentraiton and N-FENOL MIX? in 0.5ml/l and elicitor ASA were used in three different concentration: low [10-5 mol/l], medium [ 10-4 mol/l] and high [10-3 mol/l]. In the conclusion I did statistical analysis of the above montioned experiments and compared them with other Publisher experiments. Then I propřed the growing technology and utilization of Milk Thistle.

The aim of this thesis was to study the effect of the acetylsalicylic acid on the stimulation of plant immunity and thus the influence on the content of the active constituents in Silybum marianum L. plants. The main active constituents of Silybum marianum L. seeds are silybine, silydianine, isosilybine, silycristine, usually expressed as silymarine content, and taxifoline. These constituents have antihepatotoxic effect and many different protective effects on numerous organs and cells. Knowledge about stress and elicitation, origin, botanical characterization, growth, development and cultivation of Silybum marianum L. were summarized before the research.The small-plot experiment was set up in Hluboká nad Vltavou in 2013. Plants of Silybum marianum were divided into four groups and then three goups were treated three times during the vegetation with the acetylsalicylic acid of three different concentrations - 10-3 mol.l-1 (high), 10-4 mol.l-1 (medium) a 10-5 mol.l-1 (low). Every single one group was treated with only one concentration of the acetylsalicylic acid. The last group was treated only with water and served as a control group. The preparation of the extracts was being held with using mixture of acetone, methanol and water. The extracts were determined by High Performance Liquid Chromatography.The effects of each chosen concetration of acetylsalicylic acid (high, medium, low) on the active constituents in seeds were not statistically proven compared to seeds without aplication of the elicitor (control group). The ineffectiveness of the elicitor should have been also caused by nonoptimal condition of plants due to various abiotic and biotic stressors.

Thesis (Ph.D. in Toxicology) -- University of Colorado Denver, 2008. / Typescript. Includes bibliographical references (leaves 144-170). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

The liver diseases represent a relevant health problem, affecting once an organ of great importance in homeostasis maintaining. The search for herbal hepatoprotective capacity has been increasing, mainly because of the synthetic products used for this purpose may present adverse effects. Guaraná, obtained from the roasted seeds of the Amazonian plant Paullinia cupana var. sorbilis (Sapindaceae), is a product with a wide reputation as stimulant and a nutritional supplement in terms of physical and mental stress. It also presents several other biological properties, among them the antioxidant, antimicrobial, anticancer and anti-obesogenic. However, few studies focus on the potential hepatoprotective this plant. In this study, the protective effects of Paullinia cupana Mart. var. sorbilis (guaraná) on liver damage were evaluated in rats exposed to carbon tetrachloride (CCl4). Male Wistar rats were intra gastric pretreated with guaraná powder (100, 300 and 600 mg/Kg) or silymarin 100 mg/Kg daily for 14 days before treatment with a single dose of CCl4 (50% CCl4, 1 mL/Kg, intraperitoneally). Rats were sacrificed 24 h later, and blood samples were collected for assaying serum biochemical parameters. The hepatic tissue was removed to perform the comet assay. CCl4 induced liver damage and significantly increased the activities of Alanine Aminotransferase (ALT) and Aspartate Aminotransferase (AST) in serum. In addition, CCl4 increased the DNA damage index in hepatocytes. Pretreatment with guaraná in all concentrations was significantly effective in decreasing the ALT and AST activities when compared with CCl4 treated group. Furthermore, the treatment with guaraná 300 mg/Kg decreased DNA damage index. In addition, the DNA damage index showed a significant positive correlation with AST and ALT. These results indicate that guaraná has hepatoprotective activity and prevents the DNA strand breakage in CCl4-induced liver damage in rats. / As doenças hepáticas representam um grave problema de saúde, uma vez que acometem um órgão de grande importância na manutenção da homeostase. A busca por fitoterápicos com capacidade hepatoprotetora vem aumentando, principalmente pelo fato dos produtos sintéticos utilizados para esse fim apresentarem efeitos adversos. O guaraná, obtido das sementes torradas da planta amazônica Paullinia cupana var. sorbilis (Sapindaceae), é um produto com ampla reputação como estimulante e como suplemento nutricional em condições de estresse físico e mental. Além disso, apresenta outras diversas propriedades biológicas, entre estas destacam-se os efeitos antioxidantes, antimicrobianos, anticancerígenos e anti-obesogênicos. Entretanto, poucos estudos enfocam o possível potencial hepatoprotetor desta planta. Neste trabalho, os efeitos protetivos do guaraná sobre o dano hepático foram avaliados em ratos expostos ao tetracloreto de carbono (CCl4). Ratos Wistar machos foram pré-tratados (gavagem) com pó de guaraná (100, 300 e 600 mg/Kg) ou com silimaria 100 mg/Kg durante o período de 14 dias. No 14a dia, para a indução de hepatotoxidade, os animais receberam dose única, via intraperitonial de CCl4 (1 mL/Kg; 50%, diluído em azeite de oliva). Após 24 h, amostras de sangue foram coletadas via retro-orbital para avaliação de parâmetros bioquímicos, os animais sacrificados e o tecido hepático removido para a realização do ensaio cometa. O tratamento com CCl4 provocou dano hepático, demonstrado pela elevação signifivativa da atividade das enzimas aspartato aminoranferase (AST) e da alanina aminotransferase (ALT) no soro. Além disso promoveu um aumento significativo no índice de dano ao DNA. O pré-tratamento com guaraná, em todas as concentrações, foi significativamente efetivo em evitar a elevação da atividade de AST e ALT quando comparados ao grupo controle CCl4. O pré-tratamento com guaraná 300 mg/Kg promoveu uma diminuição no índice de dano ao DNA. O índice de dano no DNA mostrou signifivativa correlação positiva com as atividades de AST e ALT. Estes resultados indicam que o guaraná tem atividade hepatoprotetora e previne a quebra da fita do DNA em dano induzido por CCl4 em ratos.

L'infection par le virus de l'hépatite C (VHC) est un problème de santé publique majeur car en absence de vaccin et de thérapie suffisamment efficace, l’infection peut dégénérer en carcinome hépatocellulaire. Il est alors important d’identifier de nouvelles cibles thérapeutiques et de développer de nouveaux antiviraux. Ainsi, nous avons étudié l'activité anti-VHC de différents composés : l'arbidol (Arb), la silymarine (SM) et les molécules la composant, notamment la silibinine (SbN). Ces composés ont l'avantage d'être déjà utilisés en médecine humaine depuis de nombreuses années et ont ainsi prouvé leur innocuité. Ils présentent un large spectre antiviral et inhibent plusieurs étapes du cycle viral, dont la fusion membranaire. Cette étape du cycle est intéressante à cibler car le virus serait bloqué précocement, avant de provoquer des dommages cellulaires.Nous avons approfondi notre connaissance du mécanisme d'inhibition de la fusion par Arb en montrant par différentes stratégies qu'il s'associe avec les phospholipides à l'interface membranaire et interagit avec des résidus aromatiques. Cela suggère que Arb pourrait former durant le processus de fusion un complexe entre glycoprotéine virale et membrane, permettant d'inhiber les changements conformationnels de la glycoprotéine, nécessaires à la fusion. De même SM et ses composés inhibent la fusion de pseudoparticules de HCV, probablement en stabilisant les membranes impliquées dans le processus. Enfin, nous avons observé une activité antivirale et anti-inflammatoire très différente entre deux formulations de SbN. Tous ces résultats sont discutés dans le contexte actuel d'un arsenal thérapeutique anti-HCV qui reste limité. / Infection by the hepatitis C virus (HCV) is a major public health problem since the infection can lead to hepatocellular carcinoma in the current absence of vaccine and effective treatment. It is therefore important to identify new therapeutic targets and to develop novel antiviral drugs. Here we studied the anti-HCV activity of two compounds : arbidol (Arb), the herbal extract silymarin (SM) and molecules therein, including silibinin (SbN). These compounds are already in use in human medicine for several years and have proven safety. They display a broad antiviral spectrum and inhibit several steps of the virus life cycle, including membrane fusion. This step is very interesting to target, since the virus could be blocked upstream the cellular damages it could induce. Using different biophysical strategies, we showed that Arb associates with phospholipids at the membrane interface and interacts with aromatic residues. This suggests that Arb could form during the fusion process a complex between viral glycoprotein(s) and membrane, leading to the inhibition of the conformational changes within the glycoprotein that are required during the fusion process. SM and its components inhibit fusion of HCV pseudoparticles, probably by stabilizing the membranes involved in this process. Finally, we observed different antiviral and anti-inflammatory activities between two different formulations of SbN. Knowledge of these antiviral mechanisms should lead to innovative therapeutic strategies against HCV.