Wan, Chung Tin. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2008. / Includes bibliographical references (leaves 126-139). / Abstracts in English and Chinese. / Thesis / Assessment Committee --- p.i / Acknowledgement --- p.ii / Abstract (English) --- p.iii / Abstract (Chinese) --- p.v / List of Abbreviations --- p.vii / List of Tables --- p.ix / List of Figures --- p.x / Chapter Chanter 1 --- Introduction / Chapter 1.1. --- Overview of cancer --- p.1 / Chapter 1.1.1 --- Breast cancer --- p.1 / Chapter 1.1.2 --- Hepatocellular carcinoma (HCC) --- p.2 / Chapter 1.2. --- Role of natural products in the fight against cancer --- p.3 / Chapter 1.2.1 --- Introduction of Astragalus membranaceus --- p.4 / Chapter 1.2.2 --- Introduction of Acanthopanax senticosus --- p.9 / Chapter 1.2.3 --- Introduction of Grifola Frondosa --- p.13 / Chapter 1.3. --- Apoptosis and cancer --- p.17 / Chapter 1.3.1 --- Caspases --- p.20 / Chapter 1.3.2 --- Intrinsic apoptotic pathway --- p.21 / Chapter 1.3.3 --- Extrinsic apoptotic pathway --- p.23 / Chapter 1.3.4 --- Execution of apoptosis --- p.26 / Chapter 1.4 --- Cell cycle and cancer --- p.27 / Chapter 1.5 --- Objective of this project: --- p.29 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials: --- p.30 / Chapter 2.1.1 --- Culture medium --- p.30 / Chapter 2.1.2 --- Buffers and reagents --- p.31 / Chapter 2.1.3 --- Herbs --- p.34 / Chapter 2.2 --- Methods --- p.35 / Chapter 2.2.1 --- Cell cultures --- p.35 / Chapter 2.2.1.1 --- Cell lines --- p.35 / Chapter 2.2.1.2 --- Cell culture techniques --- p.36 / Chapter 2.2.1.3 --- Cryopreservation of cell lines --- p.36 / Chapter 2.2.2 --- Proliferation assay --- p.37 / Chapter 2.2.2.1 --- MTT assay --- p.37 / Chapter 2.2.2.2 --- Isolation of PBMC --- p.38 / Chapter 2.2.2.3 --- XTT --- p.39 / Chapter 2.2.3 --- DNA fragmentation --- p.39 / Chapter 2.2.4 --- Flow cytometry --- p.41 / Chapter 2.2.4.1 --- Detection of mitochondrial membrane depolarization by the use of JC-1 --- p.41 / Chapter 2.2.4.2 --- Annexin-V-FITC PI labeling of apoptotic cells --- p.42 / Chapter 2.2.4.3 --- Cell cycle analysis --- p.43 / Chapter 2.2.5 --- Western blotting --- p.43 / Chapter 2.2.5.1 --- Total protein extraction --- p.44 / Chapter 2.2.5.2 --- Determining protein concentration --- p.44 / Chapter 2.2.5.3 --- SDS-PAGE --- p.45 / Chapter 2.2.5.4 --- Electroblotting --- p.45 / Chapter 2.2.5.5 --- Probing --- p.46 / Chapter 2.2.5.6 --- ECL --- p.46 / Chapter 2.2.6 --- Preparation of herbal extracts --- p.47 / Chapter 2.2.6.1 --- Preparation of extract of Astragalus membranaceus and Grifola frondosa --- p.47 / Chapter 2.2.6.2 --- Preparation of Acanthopanax senticosus aqueous extract --- p.47 / Chapter 2.2.6.3 --- Partition of Acanthopanax --- p.47 / Chapter 2.2.6.4 --- Column purification of ethyl-acetate fraction --- p.48 / Chapter 2.2.6.5 --- Analytical thin layer chromatography --- p.49 / Chapter 2.2.7 --- Statistical Analysis --- p.50 / Chapter Chanter 3 --- Results / Chapter 3.1 --- Extractions --- p.51 / Chapter 3.2 --- Anti-proliferative effect of herbal extracts on cancer cell lines --- p.51 / Chapter 3.3 --- Partition of Acanthopanax methanol extract --- p.56 / Chapter 3.4 --- Anti-proliferative effect of Acanthopanax partition fractions on breast cancer cells --- p.59 / Chapter 3.5 --- Column chromatography of ethyl acetate fraction --- p.59 / Chapter 3.6 --- Anti-proliferative effect of various sub-fractions on breast cancer cells --- p.64 / Chapter 3.7 --- Effect of sub-fractions on PBMC proliferation --- p.76 / Chapter 3.8 --- Kinetic study of anti-proliferative effect of Fc --- p.76 / Chapter 3.9 --- Flow cytometric analysis --- p.79 / Chapter 3.91 --- JC-1 staining --- p.79 / Chapter 3.92 --- Annexin-P1 labeling --- p.79 / Chapter 3.93 --- Cell cycle analysis --- p.80 / Chapter 3.10 --- DNA fragmentation assay --- p.88 / Chapter 3.11 --- Western blotting --- p.91 / Chapter Chanter 4 --- Discussion --- p.99 / Chapter Chapter 5 --- Conclusion --- p.124 / References --- p.126

Orientador: Dora Maria Grassi-Kassisse / Dissertação (mestrado) - Universidade Estadual de Campinas, Instituto de Biologia / Made available in DSpace on 2018-08-21T06:42:56Z (GMT). No. of bitstreams: 1 Arouca_AlineBarbedo_M.pdf: 2816326 bytes, checksum: 7245a319bb7b00c52b0d7ae0cc6d6b02 (MD5) Previous issue date: 2012 / Resumo: O ginseng é um dos suplementos herbais mais populares do mundo. Investigações revelaram que o ginseng e seus produtos relacionados reduzem o processo inflamatório, produzem atividade adaptogênica promovendo a melhora na performance em ratos e camundongos. Das diferentes espécies de ginseng, o Siberiano (Eleutherococcus senticosus) é o que apresenta menor índice de efeitos colaterais, não produzindo excitação em pacientes, exercendo efeito protetor mais intenso sobre o sistema imune, e não desenvolvendo síndrome semelhante ao estresse, conforme já observado no Panax ginseng (ginseng Coreano). Desta forma, propomos neste trabalho, avaliar a eficácia da administração crônica de Eleutherococcus senticosus (ginseng Siberiano) sobre a performance de ratos adaptados à esteira de corrida 3 x/semana (10 m/minuto, durante 10 minutos) e submetidos a treinamento de corrida (incremento de velocidade e duração do treino até a 4ª semana, e depois mantendo 25 m/minuto, durante 60 minutos/dia até a 8ª semana), aplicando três testes de performance (teste incremental de velocidade sem inclinação) ao longo de 8 semanas de tratamento. Os resultados indicaram que não houve melhora da performance dos ratos dos grupos adaptados e corrida no segundo e no terceiro teste (meio e final do tratamento, respectivamente), e a performance do grupo corrida só foi maior que a do grupo adaptado quando os ratos foram suplementados com 100 mg/kg de peso/dia de extrato padronizado de Eleutherococcus senticosus (ES). O ganho de peso foi semelhante entre os grupos adaptado e corrida, entretanto o tratamento com o ES induziu a um menor ganho de peso dos ratos quando comparados com seus respectivos controles. A ingesta alimentar, por sua vez, foi significativamente menor no grupo Corrida e o tratamento com ES não alterou este resultado. A morfometria dos adipócitos indicou menor área dos adipócitos dos ratos suplementados em relação aos seus controles, sendo significativo no panículo epididimal para o grupo Adaptado. Em relação aos biomarcadores de estresse oxidativo, no grupo Corrida Suplementado foi observado uma tendência a menor produção de espécies reativas de oxigênio analisados pela concentração de TBARS plasmático e muscular, porém não estatisticamente diferentes. Quanto aos outros biomarcadores metabólicos (lactato e glicemia plasmáticos, e glicogênio hepático e muscular), e ao perfil lipídico (triacilglicerol e colesterol total plasmáticos), não observamos alterações significativas que demonstrassem a eficácia do tratamento com ginseng no exercício proposto. Especificamente os outros biomarcadores de estresse oxidativo (catalase e superóxido dismutase) não apontaram aumento significativo na atividade antioxidante enzimática. Concluímos, portanto, que o tratamento com Eleutherococcus senticosus na forma de tratamento proposto foi eficiente em melhorar a performance no grupo treinado em esteira (Corrida Suplementado) / Abstract: Ginseng is one of the most popular herbal supplements in the world. Investigations revealed that ginseng and its related products reduce inflammation, produce adaptogenic activity promoting improvement in performance in rats and mice. Of different species of ginseng, Siberian (Eleutherococcus senticosus) is due to a lower rate of side effects, not producing arousal in patients, exerting stronger protective effect on the immune system, and not developing like syndrome to stress, as observed in Panax ginseng (Korean ginseng). Thus, we propose in this work was to evaluate the efficacy of chronic administration of Eleutherococcus senticosus (Siberian ginseng) on the performance of rats adapted to treadmill running 3 x/week (10 m/min for 10 min) and subjected to running training (increment of speed and duration of the training until the 4th week, and then maintaining 25 m/minute for 60 minutes / day to 8 weeks) by applying three performance tests (Test incremental speed without inclination) over 8 weeks treatment. The results indicated that there was no improvement of the performance of rats in groups adapted and running in the second and third test (middle and end of treatment, respectively) and running performance only group was higher than the group adapted when the rats were treated with 100 mg/kg/day of standardized extract of Eleutherococcus senticosus (ES). Weight gain was similar between groups adapted and race, however treatment with ES induced a lower weight gain of rats when compared to their respective controls. The food intake, in turn, was significantly lower in the group racing and ES treatment did not alter this result. The adipocyte morphometry showed the smallest area of adipocytes supplemented rats compared to their controls, with significant difference in panniculus epididymal isolated of Adapted group. Regarding biomarkers of oxidative stress in Race Supplemented group was observed a trend to lower production of reactive oxygen species analyzed by plasma and muscular TBARS concentration, but not statistically different. As for other metabolic biomarkers (plasma lactate and glucose, and liver and muscle glycogen), and lipid profile (total cholesterol and plasma triacylglycerol), we observed no significant changes to demonstrate the efficacy of treatment with ginseng in the proposed exercise. Specifically other biomarkers of oxidative stress (catalase and superoxide dismutase) showed no significant increase in antioxidant enzyme activity. We therefore conclude that treatment with Eleutherococcus senticosus as proposed treatment was effective in improving performance in the group trained on a treadmill (Race Supplemented) / Mestrado / Fisiologia / Mestre em Biologia Funcional e Molecular