Fotossensibilização refere-se ao aumento da resposta à radiação ultravioleta da luz solar, causada por um agente fotodinâmico na pele. A sensibilidade à fotossensibilização de 80 cordeiros mestiços desmamados e manejados em lotação rotativa, durante 120 dias, foi avaliada em pastagem de Brachiaria brizantha cv. Marandu. Os cordeiros foram suplementados a 1% do seu peso corporal com os tratamentos, a saber: A - suplemento basal com 12% PB; B - suplemento com 18% de PB; C - suplemento com 18% de PB + propionato de Zn e D - suplemento com 18% de PB + cloreto de colina protegida. Ao término do experimento os animais foram abatidos. Os objetivos foram: avaliar o efeito da adaptação dos animais suplementados às condições de manejo em Brachiaria brizantha e a toxidez da planta aos cordeiros; caracterizar as causas da fotossensibilização; viabilizar o possível controle da verminose associada ao manejo de cordeiros em lotação rotativa e à suplementação mineral e proteica, com ou sem adição de aditivos; relacionar os teores de saponina e a presença de esporos de Pithomyces chartarum em amostras da forragem à ocorrência de casos de fotossensibilização hepatógena. O delineamento experimental foi em blocos completos e casualizados, com quatro tratamentos e quatro repetições, num total de 5 animais por tratamento. Realizou-se a análise da variância para as variáveis e as médias foram comparadas pelo teste t-Student ao nível de significância de 5%. Foram observadas baixas contagens de esporos de Pithomyces chartarum (0 - 15.000 esporos) na parte senescente da planta. A concentração de saponina protodioscina variou de 0,52 a 0,65%. Do 8º ao 58º dia ocorreram 20% de mortalidade dos animais. A avaliação histopatológica, do fígado dos animais que vieram a óbito, foi compatível com as lesões causadas pela saponina. Os padrões histológicos das lesões hepáticas encontradas nos animais, ao final do experimento, não diferiram entre os tratamentos, bem como para as análises bioquímicas: colesterol, triglicerídeos, bilirrubinas, proteína total, albumina, creatinina, ureia, AST, GGT. Ocorreram diferenças nas concentrações séricas de bilirrubinas, proteínas, albumina, AST, GGT, ureia, creatinina, colesterol, triglicerídeos, entre cordeiros sobreviventes e os que morreram. O método rotativo foi efetivo no controle da verminose, ficando a contagem de ovos por grama de fezes (OPG) abaixo de 500 ao final do experimento, não ocorrendo diferença entre os suplementos proteicos e entre os aditivos utilizados. Não houve efeito no desempenho dos cordeiros, que chegaram ao final do experimento, pela inclusão de suplementos proteicos e de aditivos. / Photosensitization refers to increase in response to ultraviolet radiation from sunlight, caused by a photodynamic agent into the skin. The sensitivity of the photosensitization at 80 crossbred lambs weaned was evaluated on Brachiaria brizantha cv. Marandu managed in rotational grazing for 120 days. Lambs were supplemented to 1% of their body weight with different treatments: A - basal supplement with 12% CP; B - supplement with 18% CP; C - supplement with18% CP + zinc proprionate and D - supplement with18 % of CP + protected choline chloride. At the end of the experiment the animals were slaughtered. The objectives were to evaluate the effect of adaptation of animals supplemented to handling Brachiaria brizantha and toxicity of the plant to lambs weaned; to characterize the causes of photosensitivity, the possible control of parasitism associated with the management of sheep grazing on rotational grazing, mineral supplementation and protein with or without the addition of additives; to relate the levels of saponin and the presence of spores Pithomyces chartarum in samples of forage and the occurrence of hepatic photosensitization. The experimental design was a complete randomized block design and assigned to four treatments and four replications for a total of five animals per treatment. The analysis of variance was performed and the means test t-Student a significance level of 5% for the effect of each treatment was used. Low count of Pithomyces chartarum spores (spores 0-15,000) in the senescent part of the plant was observed. The concentration of saponin protodioscine ranged from 0.52 to 0.65 %. Occurred between the 8º and 58º day mortality of 20 % of the animals. Histopathology of the liver of animals that died during the experiment was consistent with lesions determined by saponin. The histological patterns of liver lesions found in animals at the end of the experiment did not differ between treatments, as well as, for biochemical analysis: cholesterol, triglycerides, bilirubin, total protein, albumin, creatinine, urea, AST and GGT. There were differences in serum bilirubin, protein, albumin, AST, GGT, urea, creatinine, cholesterol and triglycerides between survivors and that died. The rotational grazing system was effective in the control of nematode parasites getting below 500 EPG at the end of the experiment, there was no difference between the protein concentrations, nor for the additives used. The additives zinc oxide and choline chloride did not improve the performance of sheep.

As plantas medicinais são consideradas pela população uma alternativa para tratamento de diversas doenças. Entre estas, destacam-se as plantas da família Verbenaceae, como as espécies do gênero Verbena. Espécies reconhecidas atualmente como gênero Glandularia foram anteriormente descritas como Verbena. Esses gêneros ainda possuem problemas de circunscrição, tendo em vista que são táxons muitos semelhantes, sendo muitas vezes difícil diferenciá-los. Espécies de Verbena são bem relatadas na medicina popular quanto a atividade anti-inflamatória e para o tratamento de doenças hepáticas. Diante disso, objetivou-se com este trabalho o isolamento de compostos presentes em espécies dos gêneros Glandularia e Verbena e a avaliação das atividades hepatoprotetora e antiquimiotáxica de espécies de Verbena utilizadas na medicina popular. Os compostos isolados foram identificados através de técnicas cromatográficas e espectroscópicas. Extratos metanólicos de V. montevidensis e V. litoralis e o iridoide brasosídeo, presente em ambos os extratos, foram avaliados quanto a atividade antiquimiotáxica realizada de acordo com o teste modificado de Boyden e quanto a atividade hepatoprotetora e citotoxicidade in vitro realizado em células HepG2, pelos métodos de MTT e vermelho neutro. De G. selloi foram obtidos 6β-OH-ipolamiida e dois derivados de crisoeriol com estruturas inéditas, seloisídeo A e seloisídeo B. Do extrato das partes aéreas de G. lobata foram isolados verbascosídeo e o iridoide griselinosídeo, que foi também isolado de V. hirta e V. bonariensis. De V. montevidensis obteve-se brasosídeo. No ensaio biológico antiquimiotáxico realizado com o extrato metanólico de V. montevidensis e com brasosídeo, seu produto isolado, todas as amostras apresentaram inibição significativa da migração leucocitária em relação ao controle. Os resultados obtidos para citotoxicidade in vitro demonstram que os extratos metanólicos e brasosídeo, não apresentaram citotoxicidade nas concentrações testadas. Esses extratos demonstraram atividade hepatoprotetora, em células HepG2, após lesão induzida por etanol. Brasosídeo protegeu as células nas menores concentrações testadas, sugerindo que a atividade hepatoprotetora dos extratos esteja relacionada ao sinergismo entre este composto e verbascosídeo. Os resultados encontrados nos ensaios biológicos corroboram o uso popular de algumas espécies de Verbena como agentes hepatoprotetor e anti-inflamatório. / Medicinal plants are considered by the population an alternative to treat various diseases. Among these, stand out plants of the family Verbenaceae, such as the species of the genus Verbena. Species currently recognized as the genus Glandularia were previously described as Verbena. These genera still have problems of circumscription, since the taxa are very similar, being often difficult to differentiate them. Verbena species are well reported in folk medicine regarding anti-inflammatory activity and for the treatment of liver diseases. The objective of this work was the isolation of compounds present in species of the genera Glandularia and Verbena and the evaluation of the hepatoprotective and antichemotactic activities of Verbena species used in folk medicine. The isolated compounds were identified by chromatographic and spectroscopic techniques. Methanolic extracts of the V. montevidensis and V. litoralis and the iridoid brasoside, present in both extracts, were evaluated for the antichemotactic activity performed according to the modified Boyden test and for the hepatoprotective activity and in vitro cytotoxicity performed in HepG2, by MTT and neutral red methods. From G. selloi 6β-OH-ipolamiide and two chrysoeriol derivatives were obtained with novel structures, selloiside A and selloiside B. From the extract of the aerial parts of G. lobata verbascoside and the iridoid griselinoside were isolated. This iridoid was also isolated from V. hirta and V. bonariensis while V. montevidensis afforded brasoside. In the antichemotactic biological assay performed with the methanolic extract of V. montevidensis and with brasoside, all the samples showed significant inhibition of the leukocyte migration in relation to the control. The results obtained for in vitro cytotoxicity demonstrate that the methanolic extracts and brasoside did not present cytotoxicity at the concentrations tested. These extracts demonstrated hepatoprotective activity in HepG2 cells after ethanol-induced injury. Brasoside protected the cells at the lowest concentrations tested, suggesting that the hepatoprotective activity of the extracts is related to the synergism between this compound and verbascoside. The results found in the biological assays corroborate the popular use of some species of Verbena as hepatoprotective and anti-inflammatory agents.

As plantas medicinais são consideradas pela população uma alternativa para tratamento de diversas doenças. Entre estas, destacam-se as plantas da família Verbenaceae, como as espécies do gênero Verbena. Espécies reconhecidas atualmente como gênero Glandularia foram anteriormente descritas como Verbena. Esses gêneros ainda possuem problemas de circunscrição, tendo em vista que são táxons muitos semelhantes, sendo muitas vezes difícil diferenciá-los. Espécies de Verbena são bem relatadas na medicina popular quanto a atividade anti-inflamatória e para o tratamento de doenças hepáticas. Diante disso, objetivou-se com este trabalho o isolamento de compostos presentes em espécies dos gêneros Glandularia e Verbena e a avaliação das atividades hepatoprotetora e antiquimiotáxica de espécies de Verbena utilizadas na medicina popular. Os compostos isolados foram identificados através de técnicas cromatográficas e espectroscópicas. Extratos metanólicos de V. montevidensis e V. litoralis e o iridoide brasosídeo, presente em ambos os extratos, foram avaliados quanto a atividade antiquimiotáxica realizada de acordo com o teste modificado de Boyden e quanto a atividade hepatoprotetora e citotoxicidade in vitro realizado em células HepG2, pelos métodos de MTT e vermelho neutro. De G. selloi foram obtidos 6β-OH-ipolamiida e dois derivados de crisoeriol com estruturas inéditas, seloisídeo A e seloisídeo B. Do extrato das partes aéreas de G. lobata foram isolados verbascosídeo e o iridoide griselinosídeo, que foi também isolado de V. hirta e V. bonariensis. De V. montevidensis obteve-se brasosídeo. No ensaio biológico antiquimiotáxico realizado com o extrato metanólico de V. montevidensis e com brasosídeo, seu produto isolado, todas as amostras apresentaram inibição significativa da migração leucocitária em relação ao controle. Os resultados obtidos para citotoxicidade in vitro demonstram que os extratos metanólicos e brasosídeo, não apresentaram citotoxicidade nas concentrações testadas. Esses extratos demonstraram atividade hepatoprotetora, em células HepG2, após lesão induzida por etanol. Brasosídeo protegeu as células nas menores concentrações testadas, sugerindo que a atividade hepatoprotetora dos extratos esteja relacionada ao sinergismo entre este composto e verbascosídeo. Os resultados encontrados nos ensaios biológicos corroboram o uso popular de algumas espécies de Verbena como agentes hepatoprotetor e anti-inflamatório. / Medicinal plants are considered by the population an alternative to treat various diseases. Among these, stand out plants of the family Verbenaceae, such as the species of the genus Verbena. Species currently recognized as the genus Glandularia were previously described as Verbena. These genera still have problems of circumscription, since the taxa are very similar, being often difficult to differentiate them. Verbena species are well reported in folk medicine regarding anti-inflammatory activity and for the treatment of liver diseases. The objective of this work was the isolation of compounds present in species of the genera Glandularia and Verbena and the evaluation of the hepatoprotective and antichemotactic activities of Verbena species used in folk medicine. The isolated compounds were identified by chromatographic and spectroscopic techniques. Methanolic extracts of the V. montevidensis and V. litoralis and the iridoid brasoside, present in both extracts, were evaluated for the antichemotactic activity performed according to the modified Boyden test and for the hepatoprotective activity and in vitro cytotoxicity performed in HepG2, by MTT and neutral red methods. From G. selloi 6β-OH-ipolamiide and two chrysoeriol derivatives were obtained with novel structures, selloiside A and selloiside B. From the extract of the aerial parts of G. lobata verbascoside and the iridoid griselinoside were isolated. This iridoid was also isolated from V. hirta and V. bonariensis while V. montevidensis afforded brasoside. In the antichemotactic biological assay performed with the methanolic extract of V. montevidensis and with brasoside, all the samples showed significant inhibition of the leukocyte migration in relation to the control. The results obtained for in vitro cytotoxicity demonstrate that the methanolic extracts and brasoside did not present cytotoxicity at the concentrations tested. These extracts demonstrated hepatoprotective activity in HepG2 cells after ethanol-induced injury. Brasoside protected the cells at the lowest concentrations tested, suggesting that the hepatoprotective activity of the extracts is related to the synergism between this compound and verbascoside. The results found in the biological assays corroborate the popular use of some species of Verbena as hepatoprotective and anti-inflammatory agents.

As plantas medicinais são consideradas pela população uma alternativa para tratamento de diversas doenças. Entre estas, destacam-se as plantas da família Verbenaceae, como as espécies do gênero Verbena. Espécies reconhecidas atualmente como gênero Glandularia foram anteriormente descritas como Verbena. Esses gêneros ainda possuem problemas de circunscrição, tendo em vista que são táxons muitos semelhantes, sendo muitas vezes difícil diferenciá-los. Espécies de Verbena são bem relatadas na medicina popular quanto a atividade anti-inflamatória e para o tratamento de doenças hepáticas. Diante disso, objetivou-se com este trabalho o isolamento de compostos presentes em espécies dos gêneros Glandularia e Verbena e a avaliação das atividades hepatoprotetora e antiquimiotáxica de espécies de Verbena utilizadas na medicina popular. Os compostos isolados foram identificados através de técnicas cromatográficas e espectroscópicas. Extratos metanólicos de V. montevidensis e V. litoralis e o iridoide brasosídeo, presente em ambos os extratos, foram avaliados quanto a atividade antiquimiotáxica realizada de acordo com o teste modificado de Boyden e quanto a atividade hepatoprotetora e citotoxicidade in vitro realizado em células HepG2, pelos métodos de MTT e vermelho neutro. De G. selloi foram obtidos 6β-OH-ipolamiida e dois derivados de crisoeriol com estruturas inéditas, seloisídeo A e seloisídeo B. Do extrato das partes aéreas de G. lobata foram isolados verbascosídeo e o iridoide griselinosídeo, que foi também isolado de V. hirta e V. bonariensis. De V. montevidensis obteve-se brasosídeo. No ensaio biológico antiquimiotáxico realizado com o extrato metanólico de V. montevidensis e com brasosídeo, seu produto isolado, todas as amostras apresentaram inibição significativa da migração leucocitária em relação ao controle. Os resultados obtidos para citotoxicidade in vitro demonstram que os extratos metanólicos e brasosídeo, não apresentaram citotoxicidade nas concentrações testadas. Esses extratos demonstraram atividade hepatoprotetora, em células HepG2, após lesão induzida por etanol. Brasosídeo protegeu as células nas menores concentrações testadas, sugerindo que a atividade hepatoprotetora dos extratos esteja relacionada ao sinergismo entre este composto e verbascosídeo. Os resultados encontrados nos ensaios biológicos corroboram o uso popular de algumas espécies de Verbena como agentes hepatoprotetor e anti-inflamatório. / Medicinal plants are considered by the population an alternative to treat various diseases. Among these, stand out plants of the family Verbenaceae, such as the species of the genus Verbena. Species currently recognized as the genus Glandularia were previously described as Verbena. These genera still have problems of circumscription, since the taxa are very similar, being often difficult to differentiate them. Verbena species are well reported in folk medicine regarding anti-inflammatory activity and for the treatment of liver diseases. The objective of this work was the isolation of compounds present in species of the genera Glandularia and Verbena and the evaluation of the hepatoprotective and antichemotactic activities of Verbena species used in folk medicine. The isolated compounds were identified by chromatographic and spectroscopic techniques. Methanolic extracts of the V. montevidensis and V. litoralis and the iridoid brasoside, present in both extracts, were evaluated for the antichemotactic activity performed according to the modified Boyden test and for the hepatoprotective activity and in vitro cytotoxicity performed in HepG2, by MTT and neutral red methods. From G. selloi 6β-OH-ipolamiide and two chrysoeriol derivatives were obtained with novel structures, selloiside A and selloiside B. From the extract of the aerial parts of G. lobata verbascoside and the iridoid griselinoside were isolated. This iridoid was also isolated from V. hirta and V. bonariensis while V. montevidensis afforded brasoside. In the antichemotactic biological assay performed with the methanolic extract of V. montevidensis and with brasoside, all the samples showed significant inhibition of the leukocyte migration in relation to the control. The results obtained for in vitro cytotoxicity demonstrate that the methanolic extracts and brasoside did not present cytotoxicity at the concentrations tested. These extracts demonstrated hepatoprotective activity in HepG2 cells after ethanol-induced injury. Brasoside protected the cells at the lowest concentrations tested, suggesting that the hepatoprotective activity of the extracts is related to the synergism between this compound and verbascoside. The results found in the biological assays corroborate the popular use of some species of Verbena as hepatoprotective and anti-inflammatory agents.

Fotossensibilização refere-se ao aumento da resposta à radiação ultravioleta da luz solar, causada por um agente fotodinâmico na pele. A sensibilidade à fotossensibilização de 80 cordeiros mestiços desmamados e manejados em lotação rotativa, durante 120 dias, foi avaliada em pastagem de Brachiaria brizantha cv. Marandu. Os cordeiros foram suplementados a 1% do seu peso corporal com os tratamentos, a saber: A - suplemento basal com 12% PB; B - suplemento com 18% de PB; C - suplemento com 18% de PB + propionato de Zn e D - suplemento com 18% de PB + cloreto de colina protegida. Ao término do experimento os animais foram abatidos. Os objetivos foram: avaliar o efeito da adaptação dos animais suplementados às condições de manejo em Brachiaria brizantha e a toxidez da planta aos cordeiros; caracterizar as causas da fotossensibilização; viabilizar o possível controle da verminose associada ao manejo de cordeiros em lotação rotativa e à suplementação mineral e proteica, com ou sem adição de aditivos; relacionar os teores de saponina e a presença de esporos de Pithomyces chartarum em amostras da forragem à ocorrência de casos de fotossensibilização hepatógena. O delineamento experimental foi em blocos completos e casualizados, com quatro tratamentos e quatro repetições, num total de 5 animais por tratamento. Realizou-se a análise da variância para as variáveis e as médias foram comparadas pelo teste t-Student ao nível de significância de 5%. Foram observadas baixas contagens de esporos de Pithomyces chartarum (0 - 15.000 esporos) na parte senescente da planta. A concentração de saponina protodioscina variou de 0,52 a 0,65%. Do 8º ao 58º dia ocorreram 20% de mortalidade dos animais. A avaliação histopatológica, do fígado dos animais que vieram a óbito, foi compatível com as lesões causadas pela saponina. Os padrões histológicos das lesões hepáticas encontradas nos animais, ao final do experimento, não diferiram entre os tratamentos, bem como para as análises bioquímicas: colesterol, triglicerídeos, bilirrubinas, proteína total, albumina, creatinina, ureia, AST, GGT. Ocorreram diferenças nas concentrações séricas de bilirrubinas, proteínas, albumina, AST, GGT, ureia, creatinina, colesterol, triglicerídeos, entre cordeiros sobreviventes e os que morreram. O método rotativo foi efetivo no controle da verminose, ficando a contagem de ovos por grama de fezes (OPG) abaixo de 500 ao final do experimento, não ocorrendo diferença entre os suplementos proteicos e entre os aditivos utilizados. Não houve efeito no desempenho dos cordeiros, que chegaram ao final do experimento, pela inclusão de suplementos proteicos e de aditivos. / Photosensitization refers to increase in response to ultraviolet radiation from sunlight, caused by a photodynamic agent into the skin. The sensitivity of the photosensitization at 80 crossbred lambs weaned was evaluated on Brachiaria brizantha cv. Marandu managed in rotational grazing for 120 days. Lambs were supplemented to 1% of their body weight with different treatments: A - basal supplement with 12% CP; B - supplement with 18% CP; C - supplement with18% CP + zinc proprionate and D - supplement with18 % of CP + protected choline chloride. At the end of the experiment the animals were slaughtered. The objectives were to evaluate the effect of adaptation of animals supplemented to handling Brachiaria brizantha and toxicity of the plant to lambs weaned; to characterize the causes of photosensitivity, the possible control of parasitism associated with the management of sheep grazing on rotational grazing, mineral supplementation and protein with or without the addition of additives; to relate the levels of saponin and the presence of spores Pithomyces chartarum in samples of forage and the occurrence of hepatic photosensitization. The experimental design was a complete randomized block design and assigned to four treatments and four replications for a total of five animals per treatment. The analysis of variance was performed and the means test t-Student a significance level of 5% for the effect of each treatment was used. Low count of Pithomyces chartarum spores (spores 0-15,000) in the senescent part of the plant was observed. The concentration of saponin protodioscine ranged from 0.52 to 0.65 %. Occurred between the 8º and 58º day mortality of 20 % of the animals. Histopathology of the liver of animals that died during the experiment was consistent with lesions determined by saponin. The histological patterns of liver lesions found in animals at the end of the experiment did not differ between treatments, as well as, for biochemical analysis: cholesterol, triglycerides, bilirubin, total protein, albumin, creatinine, urea, AST and GGT. There were differences in serum bilirubin, protein, albumin, AST, GGT, urea, creatinine, cholesterol and triglycerides between survivors and that died. The rotational grazing system was effective in the control of nematode parasites getting below 500 EPG at the end of the experiment, there was no difference between the protein concentrations, nor for the additives used. The additives zinc oxide and choline chloride did not improve the performance of sheep.

Submitted by Marcos Anselmo (marcos.anselmo@unipampa.edu.br) on 2016-04-04T17:31:47Z No. of bitstreams: 1 Marcelo Gomes de Gomes.pdf: 1949846 bytes, checksum: c4472edd9df88ec1d4f0e86bd59fafa9 (MD5) / Made available in DSpace on 2016-04-04T17:31:47Z (GMT). No. of bitstreams: 1 Marcelo Gomes de Gomes.pdf: 1949846 bytes, checksum: c4472edd9df88ec1d4f0e86bd59fafa9 (MD5) Previous issue date: 2015-03-02 / O lipopolissacarídeo (LPS) associado com a D-galactosamina (D-GalN), tetracloreto de carbono (CCl4) e o paracetamol (PCM), são modelos animais de hepatite fulminante aguda amplamente utilizados. O gama orizanol (γ-ORY) é uma mistura de 10 ésteres de ácido trans ferúlico e é encontrado no óleo do farelo de arroz. O objetivo deste estudo foi avaliar o efeito hepatoprotetor do γ-ORY nos diferentes modelos animais de hepatite fulminante aguda em camundongos. O experimento foi realizado através do tratamento com γ-ORY (50 mg/kg, por via oral, p.o.) durante uma semana, após uma hora do último tratamento foi administrado por via intraperitoneal o LPS (50 mg/kg) e D-GalN (500 mg/kg), foi utilizado silimarina como controle positivo. O γ-ORY foi administrado oralmente (50 mg/kg) durante uma semana, após 1 hora da última administração o dano foi induzido por uma única dose de CCI4 (1 mg/kg i.p.), a silimarina foi utilizada como controle positivo. Os animais receberam γ-ORY durante dois dias em duas doses (10 mg e 50 mg/kg p.o.), no terceiro dia o PCM (2 g/kg, p.o.). Após 24 horas os animais foram submetidos à eutanásia. Foram feitas as seguintes determinações bioquímicas: marcadores de estresse oxidativo (TBARS, NPSH, 4-HNE), defesas antioxidantes enzimáticas (CAT, SOD, GPx), marcadores de dano hepático (AST, ALT, GGT), marcadores de funcionalidade hepática (bilirrubina), marcadores inflamatórios (TNF-α, IL-1β), marcadores de apoptoses (Caspase 3 e 9) e análise histológica. O γ-ORY foi capaz de proteger significantemente em todas as determinações analisadas, podendo ser comparado ao um fármaco utilizado para o tratamento da hepatite fulminante aguda que é a silimarina. Portanto, sugerimos que o γ-ORY pode ser uma alternativa para o tratamento da hepatite fulminante aguda, pois ficou comprovado seu efeito hepatoprotetor está associado com sua capacidade antioxidante, anti-inflamatória e anti-apoptótica. / The lipopolysaccharide (LPS) associated with D-galactosamine (D-GalN), carbon tetrachloride (CCl4) and paracetamol (PCM) are acute fulminant hepatitis widely used animal models. The gamma oryzanol (γ-ORY) is a mixture of 10 esters of trans-ferulic acid and is found in rice bran oil. The objective was to assess the potential hepatoprotective effect of γ-ORY on mechanisms involved in different animal models of acute fulminant hepatitis in mice. The animals were divided into different groups depending on the experiment. γ-ORY treatment (50 mg/kg, per oral, p.o.) for a week after an hour of the last treatment before the administration by intraperitoneal of LPS (50 mg/kg) and D-GalN (500 mg/kg). γ-ORY was administered orally (50 mg/kg) for one week, after 1 hour of the last administration damage was induced by a single dose of CCl4 (1 mg /kg i.p.). The animals received γ-ORY for two days in two doses (10mg and 50mg /kg p.o.), on the third day PCM (2 g/ kg, p.o.). After 24 hours the animals were euthanized. The following biochemical determinations were made: non-enzymatic antioxidant defenses, defenses enzymatic antioxidants, liver damage markers, liver function markers, inflammatory markers, apoptosis markers and histological analysis. The γ-ORY was able to significantly protect in all analyzed determinations, and can be compared to a drug used for acute fulminant hepatitis. Therefore, we suggest that the γ-ORY can be an alternative for the treatment of acute fulminant hepatitis, as was proved its hepatoprotective effect is associated with its antioxidant capacity, anti-inflammatory and anti-apoptotic effect.

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.

Philosophiae Doctor - PhD / Introduction: Plant extracts and herbal preparations are often marketed as natural and safe alternatives to conventional medicines for the prevention and treatment of a variety of ailments, without proof of efficacy and safety. Cardiovascular, hematopoetic, hepatic and renal impairment resulting from the use of conventional drugs is widely acknowledged. However, there is less awareness of the potential toxicity of herbal preparations and other botanicals, many of which are widely perceived by the public as being effective and harmless, and are commonly used for self medication without supervision. In addition, potential interactions between herbal medicines and conventional drugs may compromise with patient management. In the safety evaluation of most substances, non human primates are preferred to rodent species for preclinical animal safety studies, because of their biological similarity to humans. They are regarded to be the best metabolic models for humans in a broad range of investigations. Additionally, a disadvantage of using small animal species in toxicological testing is that they require higher doses of drugs and more frequent administrations than in larger species. In light of these considerations, vervet monkeys are used here to investigate toxicity of a plant-derived triterpene, oleanolic acid. The focus is to determine effects of different concentrations of this triterpene on the cardiovascular, hematopoetic, hepatic and renal systems. Materials and methods: 12 male vervet monkeys used in this study were equally divided into four groups, i.e. three treatment groups (4, 10 and 25 mg/kg bodyweight), and one control group. Each individual in a treatment group received a specified concentration of oleanolic acid in food for 16 weeks. Monkeys in the control group received the vehicle (food) alone. Bodyweight, body temperature, respiratory rate, heart rate, systolic pressure, diastolic pressure, and mean arterial pressure were recorded from ketamine-anaethetized monkeys at baseline and every second week until week 16. In addition, blood samples were collected at baseline and every fourth week for clinical biochemistry indicators (serum electrolytes, enzymes, proteins, lipids, nitrogenous compounds, bilirubins and glucose) and hematological tests (red cell count and its indices, hemoglobin, haematocrit, white blood cell and differential count and platelet count). Results: No animal showed deviation from their normal behavioral patterns, food and water intake, was in poor health or died during and after completion of the study. The average bodyweights were not statistically significantly different between controls and the treated groups. The biphasic changes in the average body temperature of treated monkeys were similar to those seen in the control group during the first eight weeks of the study. No statistically significant differences were found in body temperature determinations between controls and the treated groups. Fluctuations observed in the respiratory rates of the treated monkeys were not statistically significantly different from that of the control group. Although not statistically significantly different from the controls, the systolic, diastolic and mean arterial pressures in the group treated with 25 mg/kg oleanolic acid were lower at week 16 compared to baseline, while those of the groups treated with 4 and 10 mg/kg oleanolic acid were relatively unchanged. Except for a reduction in systolic pressure of the control group, other blood pressure parameters were stable. Heart rates in the treated groups were not statistically significantly different from those in the controls. In all groups, except the control, high density lipoprotein concentrations were higher at week 16 compared to baseline. Fluctuations in low-density lipoprotein and total cholesterol concentrations were similar between controls and the treated groups. The triglycerides were lower at week 16 compared to baseline for all four groups. Upward trends from baseline to the end of the study were observed in creatine kinase concentrations of the controls and the groups that received 4 and 25 mg/kg. Concentrations of this enzyme were unchanged in the group that received 10 mg/kg oleanolic acid between baseline and the end of the study. No statistically significant differences were found with cholesterol, triglyceride and creatine kinase concentrations between treated groups and the controls. Serum concentrations of aspartate aminotransferase were unchanged in the controls and the groups treated with 4 and 10 mg/kg oleanolic acid, but changes in this parameter over time were statistically significantly different (P = 0.0452) from the controls in the group that received 25 mg/kg oleanolic acid. Despite wide fluctuations in the alanine aminotransferase concentrations in the groups that received 4 and 25 mg/kg oleanolic acid, no statistically significant differences were found with any of the treated groups compared to the controls. No statistically significantly different changes were seen in alkaline phosphatase activities between controls and the treated groups. Reductions in gamma-glutamyl transferase activities in the groups that received 4 and 25 mg/kg oleanolic acid were not statistically significantly different from concentrations of this enzyme in the controls. In addition, no statistically significant differences were evident between controls and the group that received 10 mg/kg oleanolic acid. There were no statistically significantly different changes in the total and conjugated bilirubin and glucose concentrations between controls and the treated groups. Fluctuations over time in the serum albumin and globulin concentrations were similar between treated groups and the controls, whereas total protein concentrations were relatively constant. Consequently, no statistically significant differences were found between controls and the treated groups. Wide fluctuations were observed in the creatinine concentrations of the groups that received 4 mg/kg oleanolic acid, while no such changes were encountered in the controls and the group that received 10 and 25 mg/kg oleanolic acid. Serum urea concentrations increased in all groups over time, except for the group that received 10 mg/kg oleanolic acid. Both urea and creatinine concentrations in the treated groups were not statistically significantly different from concentrations in the controls. Serum concentrations of sodium, chloride, potassium, calcium and magnesium and phosphate in the treated groups were not statistically significantly different from these electrolyte concentrations in the controls. Decline in red cell and hemoglobin concentrations of the controls and the group that received 25 mg/kg oleanolic acid were not statistically significantly different between these groups. In addition, no statistical significant differences were found in red cell and hemoglobin concentrations between controls and the groups that received 4 and 10 mg/kg oleanolic acid. Controls and the treated groups showed upward trends in haematocrit concentrations. Mean corpuscular volumes were statistically significantly increased; P = 0.0027 (4 mg/kg), P = 0.0010 (10 mg/kg), and P = 0.0022 (25 mg/kg), while mean corpuscular hemoglobin concentrations were statistically significantly reduced; P = 0.0017 (4 mg/kg), P = 0.0004 (10 mg/kg), P = 0.0002 (25 mg/kg) in the treated groups as compared to the controls. No statistically significant differences were evident in the concentrations of mean corpuscular hemoglobin between controls and the treated groups. White blood cell counts of the treated groups were not statistically significantly different from those of the controls throughout the study period. No statistically significant differences were found in the differential white cells and platelet counts between treated groups and the controls. Discussions: The results of this study showed that administration of oleanolic acid had no effects on the general wellbeing, bodyweights, body temperature, respiratory and heart rates, and blood pressure of vervet monkeys. A statistically significant increase in the aspartate aminotransferase activity of the group treated with 25 mg/kg oleanolic acid, together with the increase in the alanine aminotransferase levels during the same time period, might indicate oleanolic acid-induced hypersensitivity, and accordingly hepatocellular alteration. However, since serum concentrations of these enzymes returned to baseline levels, as well as the absence of variations over time in other parameters of the hepatic function, particularly alkaline phosphatase activity, it is likely that there was no underlying subacute liver disease. Serum renal function parameters also appeared to be within normal physiological limits. No pronounced changes were observed in the hematological parameters of monkeys that received oleanolic acid. Conclusion: This study's results, suggest that oleanolic acid does not produce cumulative liver enzyme alterations, and has no detrimental effects on the renal, hematopoetic and cardiovascular systems of vervet monkeys.

<p>U okviru ove disertacije su ispitane mogućnosti primene savremenih postupaka za dobijanje ekstrakata rtanjskog čaja (Satureja montana L.).<br />U ekstraktima dobijenim primenom superkritičnog ugljendioksida (različitih gustina) utvrđeno je prisustvo karvakrola kao najzastupljenije komponente. Ekstrakt sa najvećim sadržajem karvakrola je dobijen pri uslovima 350 bar i 50 &deg;C (60,82%), dok je najveći prinos karvakrola ostvaren pri pritiscima 325 i 350 bar i temperaturi 60 &deg;C, i iznosio je 2,4 g/100 g droge. Primenom ugljendioksida na većim pritiscima se ostvaruje efikasnija ekstrakcija karvakrola iz droge, kao i proizvodnja ekstrakata sa većim sadržajima karvakrola. Druge komponente prisutne u ekstraktima u znatno nižim koncentracijama su: p-cimen, borneol, trans-kariofilen, kariofilen-oksid, &gamma;-terpinen i linalool.<br />Da bi se utvrdila mogućnost unapređenja prinosa ekstrakcije superkritičnim ugljendioksidom, kao i pobolj&scaron;anja kvaliteta ekstrakata, ispitan je uticaj različitih predtretmana biljnog materijala. Ustanovljeno je da predtretman vodom najznačajnije povećava prinos ekstrakcije (za 25%), dok je ultrazvučni predtretman najadekvatniji izbor za dobijanje ekstrakata sa najvećim sadržajem karvakrola (66,46%). Međutim, najveći prinos ekstrakcije karvakrola iz 100 g droge postignut je primenom etanola i vode kao predtretmana.<br />Ekstrakti dobijeni primenom subkritične vode (uz variranje temperature i vremena ekstrakcije) su hemijski okarakterisani i radi utvrđivanja uslova ekstrakcije pri kojima se postiže najveći kvalitet ekstrakata u pogledu sadržaja polifenolnih komponenti, kao i antioksidantne aktivnosti, primenjena je metoda odzivne povr&scaron;ine (RSM). Utvrđena je prednost primene subkritične vode u odnosu na klasičnu metodu ekstrakcije u pogledu sadržaja polifenolnih komponenti i antioksidantne aktivnosti.<br />Suvi ekstrakti S. montana dobijeni spray drying tehnologijom su&scaron;enja (sa različitim koncentracijama maltodekstrina) su ispitani u pogledu fizičkih i hemijskih osobina. Takođe, radi ispitivanja mogućnosti primene u vidu funkcionalnih ili prehrambenih proizvoda, izvr&scaron;ena je senzorna analiza dobijenih suvih ekstrakata, i analizirana je njihova farmakolo&scaron;ka aktivnost u smislu ACE inhibitornog i antioksidantnog delovanja.<br />Ustanovljeno je da je najniža koncentracija maltodekstrina (10%) najadekvatnija za dobijanje suvog ekstrakta S. montana sa najvećim sadržajem polifenolnih komponenti, sadržajem etarskog ulja i u pogledu antioksidantne i ACE inhibitorne aktivnosti. Takođe, suvi ekstrakt sa 10% maltodekstrina ispunjava i ostale fizičko-hemijske parametre, kojima se osigurava kako efikasno su&scaron;enje, tako i kvalitet ekstrakta.<br />In vivo aktivnost odabranih ekstrakata S. montana ispitana je na modelu hepatotoksičnosti indukovane primenom ugljentetrahlorida. Analiziran je uticaj ekstrakata na funkciju jetre, enzimske parametre oksidativnog stresa i serumske parametre oksidativnog o&scaron;tećenja izazvanog ugljentetrahloridom i ustanovljen je hepatoprotektivni učinak ekstrakata S. montana. Takođe, ispitano je dejstvo ekstrakata na rast Ehrlich-ovih ascitnih tumora implantiranih mi&scaron;evima, kao i na biohemijske parametre oksidativnog stresa u ćelijama tumora i utvrđeno je da je vreme aplikacije ekstrakata od izuzetnog značaja.</p> / <p>This dissertation investigates the possibilities of applying contemporary processes for obtaining extracts of winter savory (Satureja montana L.).<br />Carvacrol was determined to be the most abundant component in extracts acquired by using supercritical carbon dioxide (of various densities). Extract with the richest content of carvacrol was obtained under conditions of 350 bar and 50 &deg;C (60,82%), while the highest carvacrol yield was produced under 325 bar and 350 bar pressures and at a 60 &deg;C temperature reaching 2.4 g/100 g of dry weight. By applying carbon dioxide on higher pressures, a more efficient carvacrol extraction from herbal material is performed, alongside production of extracts with higher carvacrol content. Other components in the extracts present in significantly lower concentrations are the following: p-cymen, borneol, trans-caryophyllene, caryophyllene-oxide, &gamma;-terpinene and linalool.<br />In order to recognize the possibility of improving the extraction yield produced by employing supercritical carbon dioxide and enhancing the quality of extracts, the influence of several different pre-treatments was investigated. It was determined that the water pre-treatment raises the extraction yield the most (for 25%), while the ultrasonic pre-treatment was the most adequate choice for acquiring extracts with the highest content of carvacrol (66.46%). However, the highest carvacrol extraction yield from 100 g of dry weight was achieved by applying ethanol and water as pre-treatment.<br />Extracts which were attained with subcritical water (by varying the temperature and time of extraction) were chemically characterized and Response Surface Method (RSM) was applied for the purposes of determining conditions of extractions under which the highest quality of extracts in terms of content of polyphenolic compounds and antioxidant activity is reached. The advantage of employing subcritical water over the classical method of extraction in respect of content of polyphenolic components and antioxidant activity was demonstrated.<br />Dry extracts of S. montana obtained by using the spray drying technology (with different concentrations of maltodextrine) were examined in terms of their physical and chemical characteristics. Also, in order to investigate the possibility of their use as functional or food products, sensory analysis of attained extracts was performed, and furthermore, their pharmacological activity in terms of the ACE inhibitory and antioxidant effect was analysed as well.<br />It was determined that the lowest concentration of maltodextrin (10%) was the most adequate for obtaining dry extracts of S. montana with the richest content of polyphenolic components and essential oil, and in respect of antioxidant and ACE inhibitory activities. Also, dry extract with 10% of maltodextrin meets all other physical-chemical parameters, which provide efficient drying and quality of extracts.<br />In vivo activity of selected extracts of S. montana was examined on a model of hepatotoxicity induced by applying carbon tetrachloride. The influence of extracts on liver function, enzyme parameters of oxidative stress, and serum parameters of oxidative damage caused by carbon tetrachloride was analysed and hepatoprotective effect of S. montana extracts was determined. In addition, the influence of extracts on the growth of Ehrlich ascite tumors implanted in mice was investigated, and their impact on biochemical parameters of oxidative stress in tumor cells as well, and it was determined that the time of application of extracts is of great significance.</p>

Le cancer colorectal est la deuxième cause de mortalité due au cancer en Europe et aux États-Unis. Etant donné l’efficacité limitée et la toxicité élevée des agents de chimiothérapie, de nouvelles approches sont nécessaires. Le flavanolignane silybine représente le principal constituant actif du chardon-marie (Silybum marianum). Les mécanismes moléculaires des propriétés anticancéreuses de la silybine ont été étudiés dans un modèle cellulaire de progression du cancer colorectal humain : les cellules SW480 issues d’un adénocarcinome, et leurs dérivées métastatiques les cellules SW620. Les effets chimiopréventifs de la silybine ont été étudiés dans un modèle de cancérogenèse colique induite par l’azoxyméthane chez le rat. La silybine induit une mort apoptotique avec activation de la caspase-3 dans les deux lignées. L’expression des récepteurs de mort TRAIL est augmentée, et la caspase-8 activée. Le potentiel mitochondrial est perturbé provoquant une libération du cytochrome c et une activation de la caspase-9. En plus de l’activation des voies apoptotiques extrinsèque et intrinsèque la silybine induit une réponse autophagique. La combinaison de la silybine et de TRAIL, un agent anti-cancéreux prometteur, provoque une mort cellulaire synergique dans les deux lignées. Un effet synergique est aussi observé avec la combinaison de la silybine et des inhibiteurs des histones déacétylases (HDAC) : TSA et SAHA. Dans le modèle chez le rat, la silybine réduit de 50% le nombre des lésions prénéoplasiques. En conclusion, la silybine est un agent naturel intéressant pour la prévention du cancer colorectal et dans le cadre d’une combinaison avec TRAIL/des inhibiteurs d’HDACs. / Colorectal cancer (CRC) is the second most common cause for cancer-related deaths in Europe and in the USA. Because of the limited efficacy and considerable toxicity of chemotherapeutic agents, new approaches are needed. The hepatoprotective flavonolignan silibinin is the major biologically active compound of the milk thistle (Silybum marianum).The molecular mechanisms of the anticancer properties of silibinin in CRC were studied in an in vitro model of cancer progression consisting of the adenocarcinoma cell line SW480 and its derived metastatic cell line SW620. Its chemopreventive effects were assessed in an in vivo model of azoxymethane-induced colon carcinogenesis in the rat. Silibinin induced apoptotic cell death with activation of caspase-3 in both cell lines. The expression of death receptors was upregulated, and caspase-8 was activated. The potential of the mitochondrial membrane was perturbed permitting the release of cytochrome c and the activation of caspase-9. Besides the activation of the extrinsic and the intrinsic apoptotic pathway, silibinin induced an autophagic response. Combination of silibinin and TRAIL, a promising anticancer agent selectively inducing apoptosis in cancer cells, induced synergistic cell death in both cell lines. Synergy in cell death induction was also observed by the combination of silibinin and the histone deacetylase (HDAC) inhibitors TSA and SAHA. In the preclinical model in the rat, silibinin administration was able to reduce by half the number of preneoplastic lesions present in the colon. In conclusion, silibinin is a promising natural agent for colon cancer chemoprevention and for combination therapy with TRAIL/HDAC inhibitors.