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Actin cytoskeleton regulates pollen tube growth and tropismBou Daher, Firas 04 1900 (has links)
La fertilisation chez les plantes dépend de la livraison des cellules spermatiques contenues dans le pollen à l’ovule. Au contact du stigmate, le grain de pollen s’hydrate et forme une protubérance, le tube pollinique, chargé de livrer les noyaux spermatiques à l’ovule. Le tube pollinique est une cellule à croissance rapide, anisotrope et non autotrophe; ainsi tout au long de sa croissance à travers l’apoplaste du tissu pistillaire, le tube pollinique puise ses sources de carbohydrates et de minéraux du pistil. Ces éléments servent à la synthèse des constituants de la paroi qui seront acheminés par des vésicules de sécrétion jusqu’à l’apex du tube. Ce dernier doit aussi résister à des pressions mécaniques pour maintenir sa forme cylindrique et doit répondre à différents signaux directionnels pour pouvoir atteindre l’ovule. Mon projet de doctorat était de comprendre le rôle du cytosquelette dans la croissance anisotrope du tube pollinique et d’identifier les éléments responsables de sa croissance et de son guidage. Le cytosquelette du tube pollinique est composé des microfilaments d’actine et des microtubules. Pour assurer une bonne croissance des tubes polliniques in vitro, les carbohydrates et les éléments de croissance doivent être ajoutés au milieu à des concentrations bien spécifiques. J’ai donc optimisé les conditions de croissance du pollen d’Arabidopsis thaliana et de Camellia japonica qui ont été utilisés avec le pollen de Lilium longiflorum comme modèles pour mes expériences. J’ai développé une méthode rapide et efficace de fixation et de marquage du tube pollinique basée sur la technologie des microondes. J’ai aussi utilisé des outils pharmacologiques, mécaniques et moléculaires couplés à différentes techniques de microscopie pour comprendre le rôle du cytosquelette d’actine lors de la croissance et le tropisme du tube pollinique. J’ai trouvé que le cytosquelette d’actine et plus précisément l’anneau d’actine localisé dans la partie sub-apicale du tube est fortement impliqué dans la croissance et le maintien de l’architecture du tube à travers le contrôle de la livraison des vésicules de sécrétion. J’ai construit une chambre galvanotropique qui peut être montée sur un microscope inversé et qui sert à envoyer des signaux tropistiques bien précis à des tubes polliniques en croissance. J’ai trouvé que les filaments d’actine sont impliqués dans la capacité du tube pollinique à changer de direction. Ce comportement tropistique dépend de la concentration du calcium dans le milieu de croissance et du flux de calcium à travers des canaux calciques. Le gradient de calcium établi dans le tube pollinique affecte l’activité de certaines protéines qui se lient à l’actine et dont le rôle est la réorganisation des filaments d’actine. Parmi ces protéines, il y a celles de dépolymérisation de l’actine (ADF) dont deux spécifiquement exprimées dans le gamétophyte mâle d’Arabidopsis (ADF7 et ADF10). Par marquage avec des proteins fluorescents, j’ai trouvé que l’ADF7 et l’ADF10 ont des expressions différentielles pendant la microsporogenèse et la germination et croissance du tube pollinique et qu’elles partagent entre elles des rôles importants durant ces différents stades. / Fertilization in plants depends on the delivery of the sperm cells in the pollen grain through the pollen tube to the ovule. The pollen tube is a highly anisotropic, fast growing cellular protuberance. Because the pollen tube is non autotrophic, it requires a steady supply of carbohydrates and minerals supplied by the pistil to sustain its growth. These elements serve for the synthesis of cell wall material, delivered to the site of cell wall assembly in secretory vesicles that are transported along the actin cytoskeleton and deposited at the growing apex of the tube. The tube has to resist external deformation forces in order to maintain its cylindrical shape and to respond to various directional signals in order to reach its target. My objectives were to identify the role of the cytoskeleton in the anisotropic growth of the pollen tube and to determine how the tube responds to directional cues. The cytoskeleton in the pollen tube consists of microfilaments and microtubules, both forming long filamentous elements. For in vitro growing pollen tubes, carbohydrates and growth minerals have to be added to the growth medium in specific amounts order to sustain pollen tube growth. I optimized the growth conditions of Arabidopsis thaliana and Camellia japonica pollen tubes which, in addition to pollen from Lilium longiflorum, were used as model species for my experiments. I developed a microwave based, fast and efficient fixation and labelling protocol for pollen tubes. I used pharmacological, mechanical, molecular and microscopical tools to study the role of the cytoskeleton in pollen tube growth and tropism. I found that the actin cytoskeleton, and more specifically the subapical actin fringe, plays an important role in the regulation of pollen tube growth and architecture through the controlled delivery of secretory vesicles to the growing apex. I constructed a galvanotropic chamber that can be mounted on an inverted microscope to induce controlled tropic triggers. I found that the actin cytoskeleton is also involved in the ability of the pollen tube to change its direction. This tropic behaviour was shown to be dependent on the concentration of calcium ions in the growth medium and calcium influx through calcium channels. The cytosolic calcium gradient in the pollen tube regulates the activity of various actin binding proteins that are responsible for remodelling the actin cytoskeleton. Among these proteins are two Arabidopsis gametophyte-specific actin depolymerizing factors (ADFs) that I tagged with two intrinsically fluorescent proteins. I found that ADF7 and ADF10 are differentially expressed during microsporogenesis and pollen tube germination and growth and that they likely divide important functions between them.
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Estudo químico e avaliação da atividade antioxidante de chá-verde brasileiro (camellia sinensis var. assamica) cultivar iac-259 / Components study and antioxidant activity evaluation of Brazilian green tea (Camellia sinensis var. assamica IAC-259 cultivar)Saito, Samuel Takashi January 2007 (has links)
O chá-verde brasileiro, obtido a partir das partes aéreas de Camellia sinensis var. assamica, teve sua produção recentemente implantada no Brasil devido ao emergente consumo. As principais atividades farmacológicas atribuídas a esta planta estão relacionadas à atividade antioxidante, quimiopreventiva e antitumoral. Neste trabalho foi desenvo lvido e validado método por cromatografia líquida de alta eficiência (CLAE) para avaliar o perfil dos constituintes majoritários [galato de epigalocatequina (EGCG), epigalocatequina, cafeína, galato de epicatequina (ECG) e epicatequina] do chá-verde brasileiro coletado na primavera e verão, e utilizando diferentes sistemas de extração. A resposta foi linear na faixa de 37-185 μg/ml para cafeína, 99-500 μg/ml para EGC, 20-100 μg/ml para catequina, 30-150 μg/ml para epicatequina, 150-800 μg/ml para EGCG, 20-105 μg/ml para galato de galocatequina e 40-205 μg/ml para ECG (r > 0,9999 para todos os compostos). Foi, também, avaliada a atividade antioxidante in vitro através do método fotocolorimétrico do DPPH· e do método enzimático da hipoxantina/xantina oxidase. Todos os sistemas de extração testados e seus respectivos extratos liofilizados apresentaram rendimento superior a 30%, sendo que o melhor sistema teve rendimento médio de 36,29%, e se mostrando mais eficiente na extração de EGCG e ECG. As amostras do verão apresentaram melhor atividade antioxidante em comparação às da primavera. Os teores dos componentes ECG e EGCG foram os únicos que correlacionaram com a atividade antioxidante in vitro (DPPH·). A análise estatística não mostrou diferença significativa (a = 0,05) nos teores de catequinas totais entre as estações primavera e verão. / The Brazilian green tea, produced from Camellia sinenis var. assamica, have been cultivated in Brazil recently because the rise at its consumption. The main pharmacological activities attributed to this plant are related to antioxidant activity as chemopreventive and anti-cancer agent. Herein, a new HPLC method was developed and validated to evaluate the profile of the major Brazilian green tea constituents [epigallocatechin gallate (EGCG), epigallocatechin, caffeine, epicatechin gallate (ECG) and epicatechin] between spring and summer, using different extraction systems. The response was linear over a range of 37-185 μg.mL-1 for caffeine, 99-500 μg.mL-1 for epigallocatechin, 20-100 μg.mL-1 for catechin, 30-150 μg.mL-1 for epicatechin, 150-800 μg.mL-1 for EGCG, 20-105 μg.mL-1 for gallocatechin gallate and 40- 205 μg.mL-1 for ECG (r > 0.9999 for all compounds). Likewise, the in vitro antioxidant activity was evaluated using DPPH· assay and hipoxanthine/xanthine oxidase assay. The extractors systems to produce the freeze-drying extract had presented yield up of 30%. The best system an average yield of 36.26% showed more efficient to extract EGCG and ECG. The summer samples presented better antioxidant activity when compared with spring samples. Only ECG and EGCG contents presented correlation with in vitro antioxidant activity (DPPH· assay). The statistic analysis did not show significant difference (a = 0.05) in total catechin content between spring and summer seasons.
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Estudo químico e avaliação da atividade antioxidante de chá-verde brasileiro (camellia sinensis var. assamica) cultivar iac-259 / Components study and antioxidant activity evaluation of Brazilian green tea (Camellia sinensis var. assamica IAC-259 cultivar)Saito, Samuel Takashi January 2007 (has links)
O chá-verde brasileiro, obtido a partir das partes aéreas de Camellia sinensis var. assamica, teve sua produção recentemente implantada no Brasil devido ao emergente consumo. As principais atividades farmacológicas atribuídas a esta planta estão relacionadas à atividade antioxidante, quimiopreventiva e antitumoral. Neste trabalho foi desenvo lvido e validado método por cromatografia líquida de alta eficiência (CLAE) para avaliar o perfil dos constituintes majoritários [galato de epigalocatequina (EGCG), epigalocatequina, cafeína, galato de epicatequina (ECG) e epicatequina] do chá-verde brasileiro coletado na primavera e verão, e utilizando diferentes sistemas de extração. A resposta foi linear na faixa de 37-185 μg/ml para cafeína, 99-500 μg/ml para EGC, 20-100 μg/ml para catequina, 30-150 μg/ml para epicatequina, 150-800 μg/ml para EGCG, 20-105 μg/ml para galato de galocatequina e 40-205 μg/ml para ECG (r > 0,9999 para todos os compostos). Foi, também, avaliada a atividade antioxidante in vitro através do método fotocolorimétrico do DPPH· e do método enzimático da hipoxantina/xantina oxidase. Todos os sistemas de extração testados e seus respectivos extratos liofilizados apresentaram rendimento superior a 30%, sendo que o melhor sistema teve rendimento médio de 36,29%, e se mostrando mais eficiente na extração de EGCG e ECG. As amostras do verão apresentaram melhor atividade antioxidante em comparação às da primavera. Os teores dos componentes ECG e EGCG foram os únicos que correlacionaram com a atividade antioxidante in vitro (DPPH·). A análise estatística não mostrou diferença significativa (a = 0,05) nos teores de catequinas totais entre as estações primavera e verão. / The Brazilian green tea, produced from Camellia sinenis var. assamica, have been cultivated in Brazil recently because the rise at its consumption. The main pharmacological activities attributed to this plant are related to antioxidant activity as chemopreventive and anti-cancer agent. Herein, a new HPLC method was developed and validated to evaluate the profile of the major Brazilian green tea constituents [epigallocatechin gallate (EGCG), epigallocatechin, caffeine, epicatechin gallate (ECG) and epicatechin] between spring and summer, using different extraction systems. The response was linear over a range of 37-185 μg.mL-1 for caffeine, 99-500 μg.mL-1 for epigallocatechin, 20-100 μg.mL-1 for catechin, 30-150 μg.mL-1 for epicatechin, 150-800 μg.mL-1 for EGCG, 20-105 μg.mL-1 for gallocatechin gallate and 40- 205 μg.mL-1 for ECG (r > 0.9999 for all compounds). Likewise, the in vitro antioxidant activity was evaluated using DPPH· assay and hipoxanthine/xanthine oxidase assay. The extractors systems to produce the freeze-drying extract had presented yield up of 30%. The best system an average yield of 36.26% showed more efficient to extract EGCG and ECG. The summer samples presented better antioxidant activity when compared with spring samples. Only ECG and EGCG contents presented correlation with in vitro antioxidant activity (DPPH· assay). The statistic analysis did not show significant difference (a = 0.05) in total catechin content between spring and summer seasons.
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Estudo químico e avaliação da atividade antioxidante de chá-verde brasileiro (camellia sinensis var. assamica) cultivar iac-259 / Components study and antioxidant activity evaluation of Brazilian green tea (Camellia sinensis var. assamica IAC-259 cultivar)Saito, Samuel Takashi January 2007 (has links)
O chá-verde brasileiro, obtido a partir das partes aéreas de Camellia sinensis var. assamica, teve sua produção recentemente implantada no Brasil devido ao emergente consumo. As principais atividades farmacológicas atribuídas a esta planta estão relacionadas à atividade antioxidante, quimiopreventiva e antitumoral. Neste trabalho foi desenvo lvido e validado método por cromatografia líquida de alta eficiência (CLAE) para avaliar o perfil dos constituintes majoritários [galato de epigalocatequina (EGCG), epigalocatequina, cafeína, galato de epicatequina (ECG) e epicatequina] do chá-verde brasileiro coletado na primavera e verão, e utilizando diferentes sistemas de extração. A resposta foi linear na faixa de 37-185 μg/ml para cafeína, 99-500 μg/ml para EGC, 20-100 μg/ml para catequina, 30-150 μg/ml para epicatequina, 150-800 μg/ml para EGCG, 20-105 μg/ml para galato de galocatequina e 40-205 μg/ml para ECG (r > 0,9999 para todos os compostos). Foi, também, avaliada a atividade antioxidante in vitro através do método fotocolorimétrico do DPPH· e do método enzimático da hipoxantina/xantina oxidase. Todos os sistemas de extração testados e seus respectivos extratos liofilizados apresentaram rendimento superior a 30%, sendo que o melhor sistema teve rendimento médio de 36,29%, e se mostrando mais eficiente na extração de EGCG e ECG. As amostras do verão apresentaram melhor atividade antioxidante em comparação às da primavera. Os teores dos componentes ECG e EGCG foram os únicos que correlacionaram com a atividade antioxidante in vitro (DPPH·). A análise estatística não mostrou diferença significativa (a = 0,05) nos teores de catequinas totais entre as estações primavera e verão. / The Brazilian green tea, produced from Camellia sinenis var. assamica, have been cultivated in Brazil recently because the rise at its consumption. The main pharmacological activities attributed to this plant are related to antioxidant activity as chemopreventive and anti-cancer agent. Herein, a new HPLC method was developed and validated to evaluate the profile of the major Brazilian green tea constituents [epigallocatechin gallate (EGCG), epigallocatechin, caffeine, epicatechin gallate (ECG) and epicatechin] between spring and summer, using different extraction systems. The response was linear over a range of 37-185 μg.mL-1 for caffeine, 99-500 μg.mL-1 for epigallocatechin, 20-100 μg.mL-1 for catechin, 30-150 μg.mL-1 for epicatechin, 150-800 μg.mL-1 for EGCG, 20-105 μg.mL-1 for gallocatechin gallate and 40- 205 μg.mL-1 for ECG (r > 0.9999 for all compounds). Likewise, the in vitro antioxidant activity was evaluated using DPPH· assay and hipoxanthine/xanthine oxidase assay. The extractors systems to produce the freeze-drying extract had presented yield up of 30%. The best system an average yield of 36.26% showed more efficient to extract EGCG and ECG. The summer samples presented better antioxidant activity when compared with spring samples. Only ECG and EGCG contents presented correlation with in vitro antioxidant activity (DPPH· assay). The statistic analysis did not show significant difference (a = 0.05) in total catechin content between spring and summer seasons.
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Desenvolvimento e validação de métodos analíticos para o controle de qualidade de formas farmacêuticas contendo chá verde (Camellia sinensis) e estudos de liberação e permeação cutâneaAlves, Michele Campos 12 August 2013 (has links)
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Previous issue date: 2013-08-12 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Atualmente, existe um interesse crescente nas potenciais atividades de
proteção à saúde do chá verde (Camellia sinensis), que é caracterizado pela
presença de grandes quantidades de polifenois, sendo a maioria deles
representados por catequinas. Galato de epigalocatequina (EGCG) é o componente
mais abundante e ativo, sendo geralmente utilizado como biomarcador junto com a
cafeína (CAF) e o ácido gálico (AG). EGCG é pouco absorvido no
tratogastrointestinal e está sujeito ao metabolismo de primeira passagem pelo fígado
quando administrado por via oral, forma mais comumente utilizada para o chá verde.
Um sistema transdérmico poderia então ser um mecanismo de liberação alternativo.
Dessa forma, o presente trabalho teve como objetivo: desenvolver e validar métodos
por cromatografia em fase líquida de alta eficiência (CLAE) para a quantificação
simultânea de EGCG, CAF e AG em cápsulas e emulsões contendo extrato seco de
chá verde e determinar na emulsão a liberação de EGCG in vitro e sua permeação
cutânea em pele humana, utilizando modelo ex vivo. Os métodos utilizaram CLAE
de fase inversa e detecção em arranjo de fotodiodo. A separação foi atingida
utilizando as seguintes condições: coluna octadecilsilano; fase móvel composta por
água, etanol, acetato de etila e ácido acético (84:12:3:1, v/v/v/v); temperatura do
compartimento para a coluna de 35 °C; e um sistema de fluxo por gradiente,
alternando o fluxo entre 0.7 e 1.4 mL.min-1. Os compostos foram separados em 35
min. Os métodos foram simples, seletivos, precisos, exatos e rápidos; e mostraram a
confiabilidade necessária para que fossem utilizados no controle de qualidade das
formulações testadas. O método para a emulsão provou ser adequado para os
ensaios de liberação in vitro e de permeação ex vivo. A taxa de liberação in vitro de
EGCG foi de 8896.01 mg.cm-2, seguindo o modelo de pseudo-primeira ordem,
também conhecido como modelo de Higuchi. O teste de permeação ex vivo mostrou
que EGCG não é capaz de exercer suas atividades biológicas sistemicamente
quando utilizado a partir da emulsão testada, permanecendo apenas no estrato
córneo. / Currently, there is an increased interest in potential health-protective activities
of green tea (Camellia sinensis), which is characterized by the presence of
polyphenols huge amounts, being the majority of them catechins. Epigallocatechin 3-
gallate (EGCG) is the most abundant and active component, been generally used as
biomarker together with caffeine (CAF) and gallic acid (GA). EGCG is poorly
absorbed in the gastrointestinal tract and is subject to first-pass metabolism by the
liver when administered orally, the most widely used forms of green tea. A
transdermal system could then be an alternative delivery mechanism. In this light, the
present work aimed: to develop and validate high performance liquid chromatography
(HPLC) methods for simultaneous quantification of EGCG, CAF and GA in emulsions
and capsules containing dry extract green tea and to determine in emulsion the in
vitro EGCG release and its human skin cutaneous permeation using ex vivo model.
The methods used reversed phase HPLC and photodiode array detection.
Separation was achieved using the follow conditions: octadecylsilyl column; mobile
phase composed by water, ethanol, ethyl acetate and acetic acid (84:12:3:1, v/v/v/v);
column temperature of 35 °C; and flow gradient syst em, alternating flow between 0.7
and 1.4 mL.min-1. The compounds were separated within 35 min. The methods were
simple, selective, precise, accurate and fast; and provide the reliability required for
them to be used for quality control of tested formulations. The emulsion method
proved to be suitable both for in vitro release and ex vivo permeation. EGCG in vitro
release rate was found to be 8896.01 μg.cm–2, following pseudo-first-order model,
also known as Higuchi’s model. Ex vivo permeation testing showed that EGCG is not
able to exert its biological activities systemically when used from tested emulsion,
remaining only in the stratum corneum.
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Actin cytoskeleton regulates pollen tube growth and tropismBou Daher, Firas 04 1900 (has links)
No description available.
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Geochemical Characterization of Tea Leaves (Camellia sinensis) and Soils for Provenance Studies based on Compositional Data AnalysisPospiech, Solveig 17 September 2018 (has links)
No description available.
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Efeito neuroprotetor do vinho ruby cabernet sobre o prejuízo de memória em camundongos hipercolesterolêmicos / Neuroprotective effect of ruby cabernet wine on injury memory in hypercholesterolemic miceViana, Cristini Escobar 11 April 2018 (has links)
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Previous issue date: 2018-04-11 / A hipercolesterolemia se caracteriza pela elevação patológica da taxa de colesterol sanguínea
aumentando a formação de radicais livres, o qual provoca oxidação e danos nos lipídios
celulares causando crescimento da atividade inflamatória e progressão negativa da função
cognitiva. O consumo de vinho tinto atua minimizando estes efeitos, inibindo a expressão de
citocinas e modulando os níveis do fator neurotrófico derivado do cérebro (BDNF), da
proteína cíclica de ligação ao elemento AMPc-resposta (CREB), dos microRNAs (miARN), e
a ação da enzima acetilcolinesterase (AChE). Os antioxidantes, os flavonóides e os polifenóis
tornaram-se as primeiras substâncias contidas no vinho tinto com efeitos benéficos
comprovados em várias doenças, como a inibição da oxidação de LDL e parâmetros
envolvidos na resposta inflamatória. A ingestão regular de bebidas ricas em polifenóis
demonstra exercer consequências favoráveis à saúde humana, como diminuição da incidência
de doenças cardiovasculares, câncer e proteção contra doenças neurodegenerativas, entre
outras. Portanto, o presente estudo teve como objetivo investigar o potencial efeito
neuroprotetor do vinho Ruby Carbenet em camundongos nocaute para o receptor LDLr-/-
expostos à dieta hipercolesterolêmica. O vinho foi submetido a análise em vitro para
quantificação de polifenóis, antocianinas, ácido cafeico, ácido gálico, epicatequina e
resveratrol. Foram utilizados 40 camundongos adultos com 90 dias, de 20 a 25 gramas. Os
animais foram divididos em 4 grupos (n=10) e para a indução da hipercolesterolemia foi
borrifado colesterol diluído em álcool puro 96° C, sobre a dieta comercial (Puro Trato PB 22)
durante 90 dias. Ao final do tratamento foi realizado teste comportamental de memória, tarefa
do labirinto aquático de Morris (MWMT), logo após, o sangue foi coletado por punção
cardíaca e o córtex pré-frontal e o hipocampo foram removidos para os ensaios bioquímicos.
Foram determinados os níveis de lipídios plasmáticos e as citocinas pró-inflamatórias,
interferon gama (IFN-γ), fator de necrose tumoral-α (TNF-α), interleucina-1β (IL-1β) e
interleucina-6 (IL-6) no hipocampo e córtex pré-frontal. Também foram avaliadas a atividade
da AChE e os níveis das proteínas estruturais BDNF e CREB, e os biomarcadores
microRNAs. Nossos resultados mostraram que a dieta hipercolesterolemica causa um
aumento nos lipídios plasmáticos, nas citocinas pró-inflamatórias e na atividade da enzima
AChE nos camundongos LDLr-/-, o que foi atenuado pelo consumo de vinho tinto através da
diminuição do estado inflamatório. A dieta com elevado índice de colesterol diminuiu os
níveis de BDNF, CREB e microRNAs cerebrais, e o consumo moderado de vinho tinto foi
capaz de reverter estas proteínas estruturais BDNF e CREB, e os biomarcadores miRNA146a
e 155, promovendo a neuroproteção e a melhora da memória. O vinho Ruby Cabernet
apresentou potencial antioxidante confirmado pelos seus teores de ácido gálico 135, 3 mg/L,
ácido caféico 91,2 mg/L, epicatequina 242,2 mg/L e resveratrol 102,2 mg/L. O consumo
moderado do vinho proporcionaram aos camundongos melhora no déficit cognitivo e
plasticidade, corroborando com o resultado do teste comportamental, no qual o MWMT está
relacionado com o sistema de aprendizado e memória. Com isso, verificou-se que o vinho
Ruby Carbenet demonstrou efeito neuroprotetor e reduziu os níveis dos lipídios plasmáticos,
modulando as citocinas, agindo na manutenção da neuroplasticidade e regulação da atividade
da acetilcolinesterase em camundongos hipercolesterolêmicos.
Palavras-chave: Compostos fenólicos. Déficit cognitivo. Dieta com elevado índice de
colesterol. Camundongos nocaute para receptor LDLr-/-. Neuroplasticidade. / Hypercholesterolemia is characterized by the pathological elevation of the blood cholesterol
level, increasing the formation of free radicals, which causes oxidation and damage to the
cellular lipids causing inflammatory activity growth and negative progression of cognitive
function. Red wine consumption minimizes these effects by inhibiting cytokine expression
and modulating levels of brain-derived neurotrophic factor (BDNF), cyclic cAMP-binding
element (CREB), microRNAs (miRNA), and the action of the enzyme acetylcholinesterase
(AChE). Antioxidants, flavonoids and polyphenols have become the first substances
contained in red wine with proven beneficial effects in various diseases, such as the inhibition
of LDL oxidation and parameters involved in the inflammatory response. Regular ingestion of
polyphenol-rich beverages has a positive effect on human health, such as reducing the
incidence of cardiovascular diseases, cancer and protection against neurodegenerative
diseases, among others. Therefore, the present study aimed to investigate the potential
neuroprotective effect of Ruby Carbenet wine in knockout mice for the Ldlr-/- receptor
exposed to the hypercholesterolemic diet. The wine was submitted to in vitro analysis for
quantification of polyphenols, anthocyanins, caffeic acid, gallic acid, epicatechin and
resveratrol. 40 adult mice with 90 days, from 20 to 25 grams were used. The animals were
divided into 4 groups (n=10) and for the induction of hypercholesterolemia cholesterol was
diluted in pure alcohol 96 ° C, on the commercial diet (Puro Trato PB 22) for 90 days. At the
end of the treatment was performed memory behavioral test, Morris water maze task
(MWMT), soon after, blood was collected by cardiac puncture and the prefrontal cortex and
hippocampus were removed for biochemical assays. The levels of plasma lipids and
proinflammatory cytokines, interferon gamma (IFN-γ), tumor necrosis factor-α (TNF-α),
interleukin-1β (IL-1β) and interleukin-6 (IL-6) in the hippocampus and prefrontal cortex. We
also evaluated the activity of AChE and the levels of the structural proteins BDNF and CREB,
and the biomarkers microRNAs. Our results showed that the hypercholesterolemic diet causes
an increase in plasma lipids, proinflammatory cytokines and AChE enzyme activity in Ldlr-/-
mice, which was attenuated by the consumption of red wine through the reduction of the
inflammatory state. The high cholesterol diet decreased levels of BDNF, CREB and brain
microRNAs, and moderate consumption of red wine was able to revert these structural
proteins BDNF and CREB, and the miRNA146a and 155 biomarkers, promoting
neuroprotection and the improvement of memory. Ruby Cabernet wine showed antioxidant
potential confirmed by its content of gallic acid 135.3 mg/L, caffeic acid 91.2 mg/L,
epicatechin 242.2 mg/L and resveratrol 102.2 mg/L. The moderate consumption of wine gave
the mice an improvement in cognitive deficit and plasticity, corroborating with the result of
the behavioral test, in which MWMT is related to the learning and memory system. Thus, it
was verified that Ruby Carbenet wine demonstrated neuroprotective effect and reduced
plasma lipid levels, modulating cytokines, acting to maintain neuroplasticity and regulate
acetylcholinesterase activity in hypercholesterolemic mice.
Keywords: Phenolic compounds. Cognitive deficit. Diet with high cholesterol. Knockout
mice for Ldlr-/- receptor. Neuroplasticity.
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Effect of tea and herbal infusions on mammalian reproduction and fertilityOpuwari, Chinyerum Sylvia January 2013 (has links)
<p>Camellia sinensis (tea) and Aspalathus linearis (rooibos) may improve reproductive function owing to their antioxidant properties. To test this<br />
hypothesis, male and female rats were given 2% and 5% green tea (Gt), black tea (Bt), unfermented rooibos (Ur) or fermented rooibos (Fr) as sole source of drinking for 52 and 21 days respectively. Control rats received tap water. In addition, TM3 Leydig cells were exposed to 0.025, 0.05, 0.1 and 0.5 % aqueous extracts of green tea, black tea, unfermented and fermented rooibos for 24h. In vitro analysis of tea and the herbal infusion revealed the phenolic property and antioxidant capacity (FRAP) in the order Gt > / Bt > / Ur > / Fr. Camellia sinensis and Aspalathus linearis revealed no significant effect on serum antioxidant capacity (p > / 0.05) and lipid peroxidation (MDA) in the kidney or liver in both male and female rats and in the testes of the male rats (p > / 0.05). In addition, the antioxidant levels were maintained in the testes, liver and kidneys in both the male and female rats. In the male rats, no significant alterations were observed in body weight gain, liver and reproductive organs weight, and serum testosterone (p > / 0.05). Only, 5% green tea significantly increased testosterone level (p < / 0.05). Seminiferous tubules displayed complete spermatogenesis with abundant sperm in the lumen in all treated groups. However, a significant decrease in diameter and germinal epithelial height of these tubules were observed (p < / 0.05). In the epididymides, epithelial height of caput region showed a significant increase (p < / 0.01), while the cauda region was increased by Camellia sinensis but decreased by Aspalathus linearis. Sperm concentration improved significantly by green tea and unfermented rooibos (p < / 0.05), while black tea and fermented rooibos produced a non significant effect (p > / 0.05). Sperm viability was enhanced in all treatment groups (p < / 0.05). Furthermore, green tea, black tea and unfermented rooibos significantly improved the motility of rat sperm (p < / 0.05) / fermented rooibos tended to improve it (p > / 0.05). In addition, green tea, black tea and fermented rooibos enhanced acrosome reaction (p < / 0.05). Creatinine activity was significantly higher in rats treated with black tea, unfermented rooibos or fermented rooibos (p < / 0.05), green tea tended to increase it (p > / 0.05) reflecting the significant increased kidney weight in the treatment groups at high concentrations. Liver markers, ALT and AST, decreased significantly in all treated groups (p < / 0.05), except in 5% fermented rooibos where a significant increase in AST level was observed (p < / 0.01). In the female rats, the body weight gain, and reproductive organs weight was no affected (p > / 0.05). However, 5% fermented rooibos reduced the ovarian weight (p < / 0.05), while 5% unfermented rooibos significantly increased the uterine weight (p < / 0.05). Liver weight increased significantly by black tea and unfermented rooibos (p < / 0.05) while the kidney weight increased significantly by 5% black tea (p < / 0.05). No significant effect was observed in the level of FSH produced, on the other hand, Camellia sinensis significantly lowered the level of LH (p < / 0.05), while Aspalathus linearis had no effect (p > / 0.05). Creatinine activity was enhanced significantly only by 5% fermented rooibos (p < / 0.05). Liver markers, ALT and AST were reduced in most treated groups except in fermented rooibos where an increase was observed. In addition, histological sections revealed no obvious alteration in the ovaries, uteri, kidneys and liver of all treated female rats. Camellia sinensis and Aspalathus linearis significantly reduced the level of testosterone produced in TM3 Leydig cells under stimulated conditions in vitro (p< / 0.05). Furthermore, both plants maintained the viability and morphology of the cells. However, at 0.5% of either plant extracts, a significant decrease in the viability (p < / 0.05) and altered morphology of the TM3 Leydig cells was observed. In conclusion, Camellia sinensis and Aspalathus linearis significantly improved certain sperm function which might be attributed to their high level of antioxidant activity. However, the prolonged exposure of both plant extracts might result in subtle structural changes in the male reproductive system and impair kidney function. In addition, fermented rooibos at high concentration may also impair the functions of the liver. In vitro, both plants were shown to possess anti-androgenic property on TM3 Leydig cells. Furthermore, both Camellia sinensis and Aspalathus linearis may be classified as weak phytoestrogens due to the changes in the weight of the uterus and ovaries observed.</p>
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70 |
Effect of tea and herbal infusions on mammalian reproduction and fertilityOpuwari, Chinyerum Sylvia January 2013 (has links)
<p>Camellia sinensis (tea) and Aspalathus linearis (rooibos) may improve reproductive function owing to their antioxidant properties. To test this<br />
hypothesis, male and female rats were given 2% and 5% green tea (Gt), black tea (Bt), unfermented rooibos (Ur) or fermented rooibos (Fr) as sole source of drinking for 52 and 21 days respectively. Control rats received tap water. In addition, TM3 Leydig cells were exposed to 0.025, 0.05, 0.1 and 0.5 % aqueous extracts of green tea, black tea, unfermented and fermented rooibos for 24h. In vitro analysis of tea and the herbal infusion revealed the phenolic property and antioxidant capacity (FRAP) in the order Gt > / Bt > / Ur > / Fr. Camellia sinensis and Aspalathus linearis revealed no significant effect on serum antioxidant capacity (p > / 0.05) and lipid peroxidation (MDA) in the kidney or liver in both male and female rats and in the testes of the male rats (p > / 0.05). In addition, the antioxidant levels were maintained in the testes, liver and kidneys in both the male and female rats. In the male rats, no significant alterations were observed in body weight gain, liver and reproductive organs weight, and serum testosterone (p > / 0.05). Only, 5% green tea significantly increased testosterone level (p < / 0.05). Seminiferous tubules displayed complete spermatogenesis with abundant sperm in the lumen in all treated groups. However, a significant decrease in diameter and germinal epithelial height of these tubules were observed (p < / 0.05). In the epididymides, epithelial height of caput region showed a significant increase (p < / 0.01), while the cauda region was increased by Camellia sinensis but decreased by Aspalathus linearis. Sperm concentration improved significantly by green tea and unfermented rooibos (p < / 0.05), while black tea and fermented rooibos produced a non significant effect (p > / 0.05). Sperm viability was enhanced in all treatment groups (p < / 0.05). Furthermore, green tea, black tea and unfermented rooibos significantly improved the motility of rat sperm (p < / 0.05) / fermented rooibos tended to improve it (p > / 0.05). In addition, green tea, black tea and fermented rooibos enhanced acrosome reaction (p < / 0.05). Creatinine activity was significantly higher in rats treated with black tea, unfermented rooibos or fermented rooibos (p < / 0.05), green tea tended to increase it (p > / 0.05) reflecting the significant increased kidney weight in the treatment groups at high concentrations. Liver markers, ALT and AST, decreased significantly in all treated groups (p < / 0.05), except in 5% fermented rooibos where a significant increase in AST level was observed (p < / 0.01). In the female rats, the body weight gain, and reproductive organs weight was no affected (p > / 0.05). However, 5% fermented rooibos reduced the ovarian weight (p < / 0.05), while 5% unfermented rooibos significantly increased the uterine weight (p < / 0.05). Liver weight increased significantly by black tea and unfermented rooibos (p < / 0.05) while the kidney weight increased significantly by 5% black tea (p < / 0.05). No significant effect was observed in the level of FSH produced, on the other hand, Camellia sinensis significantly lowered the level of LH (p < / 0.05), while Aspalathus linearis had no effect (p > / 0.05). Creatinine activity was enhanced significantly only by 5% fermented rooibos (p < / 0.05). Liver markers, ALT and AST were reduced in most treated groups except in fermented rooibos where an increase was observed. In addition, histological sections revealed no obvious alteration in the ovaries, uteri, kidneys and liver of all treated female rats. Camellia sinensis and Aspalathus linearis significantly reduced the level of testosterone produced in TM3 Leydig cells under stimulated conditions in vitro (p< / 0.05). Furthermore, both plants maintained the viability and morphology of the cells. However, at 0.5% of either plant extracts, a significant decrease in the viability (p < / 0.05) and altered morphology of the TM3 Leydig cells was observed. In conclusion, Camellia sinensis and Aspalathus linearis significantly improved certain sperm function which might be attributed to their high level of antioxidant activity. However, the prolonged exposure of both plant extracts might result in subtle structural changes in the male reproductive system and impair kidney function. In addition, fermented rooibos at high concentration may also impair the functions of the liver. In vitro, both plants were shown to possess anti-androgenic property on TM3 Leydig cells. Furthermore, both Camellia sinensis and Aspalathus linearis may be classified as weak phytoestrogens due to the changes in the weight of the uterus and ovaries observed.</p>
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