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Resumo: Atualmente, tem se verificado um grande avanço científico envolvendo os estudos químicos, farmacológicos e microbiológicos de plantas medicinais que visam obter novos compostos com propriedades terapêuticas. A demonstração de notável atividade antimicrobiana de extratos aquoso e hidroalcoólico de Araçá (Psidium cattleianum) sobre a microbiota bucal e a busca de substâncias que reúnam tais propriedades resguardando os princípios biológicos, nos levam a considerar uma possível utilização clínica desse extrato como coadjuvante na terapia odontológica. Desta forma, o objetivo deste estudo foi avaliar a resposta biológica imediata e tardia de soluções hidroalcoólica e aquosa preparadas à base de extrato vegetal de Araçá (Psidium cattleianum). Para a análise edemogênica (reação imediata), foram utilizados 18 ratos machos. Sob anestesia geral, os animais receberam injeção intravenosa de Azul de Evans 1%. Após 30 minutos, foi injetado 0,1mL de um dos extratos ou do controle (soro fisiológico), na região subcutânea dorsal do animal. Os animais foram sacrificados após 3 e 6 horas e as peças obtidas colocadas em formamida por 72 horas. A leitura foi realizada em espectrofotômetro com comprimento de onda de 630hm. Para a análise morfológica (reação tardia), 30 ratos receberam implantes de tubos de polietileno contendo os extratos ou o soro na região dorsal, e sacrificados após 7 e 28 dias. As peças foram processadas, cortadas e coradas com Hematoxilina e Eosina. Os resultados obtidos através da leitura dos espécimes em microscópio óptico, em aumentos de 10 e 40x, para avaliação da espessura da cápsula fibrosa e contagem de células inflamatórias, que quantificam o infiltrado inflamatório. / Abstract: Nowadays, it's been seen great scientific advance involving chemistry, pharmacological and micro biotical researches in medicinal plants. These researches can find new components with therapeutic property. The demonstration of great anti micro biotic activity of watery vegetable and hydro alcoholic extract of araça (Psidium cattleianum) on the oral microbiota and the search form substances which can put together these properties keeping the biological principles, make us consider a possible clinical use of this extract as supporting on dentistry therapy. This way, the objective of this paper (research) was evaluate the biological immediate and late answer of hydro alcoholic and watery solutions prepared with vegetable extract of araça (Psidium cattleianum). For the edemogenic analysis (immediate reaction), we used 18 male mice. Under general anesthesia, the animals received intra shot of Evans blue 1%. 30 minutes later, it was injected 0,1mL of one of the extracts or the control (physiological serum), on the dorsal under skin region of the animal. The animals were sacrificed after 3 and 6 hours and the obtained material were put in formamide for 72 hours. The reading was made for spectrophotometer with waves length of 630nm. For the morphological analysis (late reaction), 30 mice received duct implant of polyethylene with the extracts or the serum on the dorsal region, and sacrificed after 7 and 28 days. The material were processed, cut and printed with HE. The obtained results from the reading of the species in optical microscopy, with raisings of 10 and 40x, for the thickness evaluation of the fiber capsule and counting of the damaged cells, that quantified the damaged infiltrate. / Orientador: Eloi Dezan Júnior / Coorientador: Elerson Gaetti-Jardim Júnior / Banca: Sônia Regina Panzarini Barioni / Banca: Thais Marchini de Oliveira / Mestre

Diabetes mellitus is a metabolic disease characterised by hyperglycaemia resulting from defects in insulin secretion, insulin action, or both. The leaf extract of Leonotis leonurus and its active compound marrubiin, have been shown to possess anti-diabetic, antiplatelet, anti-inflammatory and anti-coagulation activity. In the present study, the mechanism by which L. leonurus and marrubiin exert their anti-diabetic properties, the cross-talk between the peripheral tissues and a nano drug delivery system were investigated. Marrubiin in the plant extract was effectively quantified by an optimised reversed phase highperformance liquid chromatography (HPLC) protocol using a pentafluorophenyl (PFP) column with water and acetonitrile (50:50) as mobile phase, and a flow rate of 1ml/min. The chemical structure was determined using liquid chromatography-tandem mass spectroscopy LC-MS/MS. Real-time quantitative polymerase chain reaction (RT-qPCR) gene expression of selected adipokines and proteins implicated in Type-2 diabetes (T2D) were investigated in specific peripheral tissues isolated from an in vivo obese rat model. An in vitro cell culture model was used to determine the crosstalk between the peripheral tissues and pancreatic (INS-1E) β-cells. Various nanoformulations of L. leonurus extract were prepared and their effect on cytotoxicity (in Chang liver and INS-1 cells), insulin-mediated glucose uptake (Change liver cells) and insulin secretion (INS-1) were investigated. The average yield of marrubiin from the plant extract was 10% (n=3), with a molecular mass of 333.20Da and a molecular formula of C20H29O4 +. Results from the in vivo study showed that the L. leonurus extract significantly (p<0.05) enhanced the gene expression of adiponectin, peroxisome proliferator-activated receptor gamma (PPAR-γ), glucokinase (GK), uncoupling protein-2 (UCP-2) and reduced leptin in adipose tissue, but resistin, glucose transporters (GLUT), fatty acid synthase (FAS), insulin receptor substrate -1 (IRS-1) and phosphoenolpyruvate carboxykinase (PEPCK) gene expression were not affected. Marrubiin decreased gene expression of leptin and resistin, and increased IRS-1 and glucokinase in adipose tissue. In liver and muscle tissues, marrubiin and the L. leonurus extract reduced gene expression of PPAR-γ, IRS-1, glucokinase and PEPCK. In the in vitro crosstalk study (under normoglycaemic and hyperglycaemic conditions), conditioned medium from 3T3-L1 cells significantly (p<0.01) enhanced insulin secretion. This was not observed in INS-1E cells exposed to muscle- and liver-conditioned medium, respectively. The in vitro studies using a nanostructured lipid formulation (NLC) of the plant extract was not cytotoxic to either INS-1 and Chang liver cells. The NLC formulation significantly (p<0.05) enhanced glucose uptake in Chang liver cells and improved chronic insulin release in INS-1 cells (p<0.05). Based on the above findings from the in vivo and in vitro studies, both L. leonurus and marrubiin exerted an insulinotropic effect via adipose tissue on pancreatic β-cells. The findings in the in vivo study showed that marrubiin and the L. leonurus extract were employing their major anti-diabetic action via the adipose tissue.

Six plant species (Aloe ferox, Aloe marlothii, Clerodendrum glabrum, Jatropha curcas, Ricinus communis and Strychnos madagascariensis) used by communities in the northern parts of South Africa to control ticks on livestock were selected from the database of the Onderstepoort Veterinary Institute. The leaves of these species were extracted with organic solvents ranging from polar to non-polar (methanol, acetone and dichloromethane). Infusions (soap-water-paraffin) and decoctions traditionally used were also prepared. The tick repelling and toxicity activities of the extracts were evaluated against the livestock tick, Rhipicephalus appendiculatus. Initial results of the tick-climbing repellency bioassay illustrated that a 30% concentration of crude acetone extracts of A. ferox, A. marlothii and C. glabrum; DCM extracts of R. communis and A. marlothii and MeOH extract of J. curcas repelled the ticks to a statistically significant level. Of all the aqueous extracts only the decoction of S. madagascariensis repelled ticks significantly. At lower concentration of 10%, only acetone extracts of C. glabrum were effective. Three types of bioassays were conducted for toxicity i.e., dipping, topical and dry extract applications. Organic extracts were not effective. Infusions of A. ferox and S. madagascariensis. had strong topical application toxicity at a concentration of 35.5 % while the same concentration of infusions of A. ferox and J. curcas had a strong dipping toxicity effect. Both A. ferox and S. madagascariensis extracts were still toxic at the concentration of 30% in topical and dipping bioassays. In general acetone was a good extractant with lower yields but higher percentage repellency. Polar aqueous extracts had high yields but lower percentage repellencies. It may be that volatiles present in organic extracts are repellents and non-volatiles from aqueous extracts are toxic. Some of the fractions of C. glabrum had a higher tick repellency activity than the commercial tick control agents used as positive controls. Unfortunately the activity decreased after 2.5 hours probably due to a volatility of the biologically active compound. Attempts were made to isolate the repellent compound from the acetone extract of C. Glabrum. The process gave very good results up to a late stage of the bioassay guided fractionation. At that stage the repellent activity was lost. When two fractions were combined the repellent activity was present again. This is strong evidence for the existence of a synergistic effect. Further attempt to isolate the compounds at this stage and characterize them by nuclear magnetic resonance spectroscopy failed probably through decomposition of the compounds. This aspect can probably be followed up in more detail. The results substantiated the rationale behind the use of these plant extracts to control ticks. Even a simple acetone extract of leaves of some species has excellent tick repelling activities. If the duration of this effect could be increased it could very well develop into a commercially useful product. The tick repelling activity of S. madagascariensis decoctions and tick killing activity of A. ferox soap infusion holds great promise for the control of ticks by rural farmers. In vivo trials under controlled conditions should provide useful results. / Dissertation (MSc)--University of Pretoria, 2008. / Paraclinical Sciences / unrestricted

published_or_final_version / Biological Sciences / Doctoral / Doctor of Philosophy

Thesis (MSc (Paravet. Studies))--University of Pretoria, 2007. / Includes bibliographical references. Also available as hard copy.

Made available in DSpace on 2014-06-11T19:25:25Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-02-26Bitstream added on 2014-06-13T20:32:57Z : No. of bitstreams: 1 vieira_ld_me_botib.pdf: 650379 bytes, checksum: 2e951ffd593a61538c5b43bfddd72a7a (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / As úlceras pépticas são lesões provocadas pelo desequilíbrio entre fatores lesivos e protetores das mucosas gástrica e duodenal. Existe um grande número de plantas que são empregadas popularmente para o tratamento deste tipo de doença. Com este trabalho, pretende-se contribuir para a compreensão da ação farmacológica dos extratos de duas dessas plantas: a Alchornea glandulosa e a Davilla elliptica, assim como a Davilla nitida, que não é empregada na etnofarmacologia, mas pertence ao mesmo gênero de plantas com a função gastroprotetora. Neste sentido, foi realizada uma análise da composição multielementar destes compostos utilizando a técnica analítica de fluorescência de raios-X por reflexão total (TXRF). Foram analisadas, também, amostras de mucosas gástricas de ratos tratados com estes extratos fitoterápicos e que tiveram úlceras pépticas induzidas por dois protocolos: indução por etanol ou indução por Indometacina, uma droga anti-inflamatória não esteroidal (DAINE). As análises exploratórias e qualitativas foram efetuadas no Laboratório de Instrumentação Nuclear do Centro de Energia Nuclear na Agricultura (CENA/USP). A determinação das concentrações dos elementos P, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br e Rb foi efetuada no Laboratório Nacional de Luz Síncrotron (LNLS) / Peptic ulcers are lesions caused by the disequilibrium between harmful and protective factors of gastric and duodenal mucosa. There are several plants that are commonly used for the treatment of this disease. The purpose of this work is contribute for the comprehension of the pharmacological action of extracts of two types of these plants: Alchornea glandulosa and Davilla elliptica, as Davilla nitida, which is not used in the ethnopharmocology, but it belongs to the same genus of plants, with gastroprotective activity. So, we have done an analysis of the multielemental composition of these compounds using the analytical technique of total reflection X-ray fluorescence (TXRF). We also analyzed samples of gastric mucosa of rats treated with herbal extracts, which had peptic ulcers induced by two protocols: induction by ethanol or by indomethacin, a non-steroidal anti-inflammatory drug (NSAID). The exploratory and qualitative analyzes were performed at the Nuclear Instrumentation Laboratory of the Brazilian Center for Nuclear Energy and Agriculture (CENA / USP). The concentrations of elements P, S, Cl, K, Ca, Ti, Cr, Mn, Fe, Ni, Cu, Zn, Se, Br and Rb were determinated at the Brazilian Synchrotron Light Laboratory (LNLS)

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