Discovery of cheap, nontoxic and readily available antiatherosclerotic drugs is an extraordinary challenge in this modern world. Atherosclerosis and cardiovascular diseases have been predicted to be the leading cause of death by the year 2030. Hence, this thesis was designed to search for plant (s) with anti-atherogenic properties, investigate its possible side effects and extrapolate its likely mechanism(s) of action. An ethnobotanical survey was employed in identification of locally important plants used for the management and treatment of cardiovascular diseases and its predisposing factors in Nkonkobe Municipality, Eastern Cape in South Africa. Information on the names of plants, their parts used and methods of preparation was collected through a questionnaire which was administered to herbalists, traditional healers and rural dwellers. The most frequently used plant (Rhizomes of Tulbaghia violacea Harv.) was investigated for toxicity using brine shrimp lethality (in vitro) and in vivo toxicity test (acute and subchronic) on rats to determine safety dosage. The in vitro antioxidant and free radical scavenging activity of the plant was investigated using models such as 1,1-diphenyl-2- picrylhydrazyl (DPPH), superoxide anions, hydrogen peroxide (H2O2), nitric oxide (NO), 2,2’- azinobis [3-ethylbenzothiazoline-6-sulfonic acid] diammonium salt (ABTS), lipid peroxidation inhibition and the ferric reducing agent. Phytochemical content and the effect of oral administration of fresh methanolic extract rhizomes of Tulbaghia violacea (250, 500 mg/kg. bwt/day) on Lipid peroxidation (TBARS), serum and tissue antioxidant enzymes in normal, hypercholesterolemic and diet induced atherogenic rats were also assessed. More so, the potential of the extract (250 and 500 mg/kg. bwt) to protect against atherogenic diet (4 percentage cholesterol 1 pecentage cholic acid and 0.5 percentage thiouracil) induced fatty streaks formation, dyslipidemia, oxidative stress and endothelial dysfunction was also investigated. Ethnobotanical study revealed that 19 plant species are used for the treatment of heart related diseases in the Municipality. 53 percentage of the plants mentioned were used for the management of chest pain, 47 percentage for high blood pressure, 42 percent for heart disease, 16 percentage for stroke and 11 percentage for the treatment of hypercholesterolemia. Tulbaghia violacea was repeatedly mentioned as the plant species used for the treatment of high blood pressure and predisposing factors in the study area. The brine shrimp cytotoxicity test revealed that fresh, dried methanolic extracts and essential oil of the T. violacea exhibited a high degree of cytotoxic activity with IC50 values of 18.18 (fresh) and 19.24 (dried) μg/ml. An IC50 value of 12. 59 μg/ml was obtained for the essential oil of the plant. The low cytotoxicity values obtained, suggested that rhizome of T. violacea may serve as a potential source of antimicrobial and anticancer agents. In vivo acute study of single oral administration of 5g/kg dose does not produce mortality or significant behavioral changes during 14 days observation. In the sub-chronic study, the extract (250, 500 mg/kg/bwt/ day) administered for a period of 28 days showed no mortality or morbidity. The weekly body and organ weight of the rats showed no significant differences between the control and the rats treated with the extract. The extract at all doses does not show any effect on of biomarkers of liver or renal damage. However, a significant decrease in the activity of ƔGT was observed in the extract treated groups. Hematological evaluation revealed that oral administration of fresh methanolic extracts of rhizomes of T. violacea does not cause anaemia or leucocytosis in the animals. Furthermore, histopathology results of the internal organs revealed no detectable inflammation. These results demonstrated that the rhizome extract of T. violacea was potentially safe for consumption orally even in chronic concentration. In vitro antioxidant evaluation showed that the essential oil, fresh and dried methanolic extracts exhibited potent antioxidant activities in a concentration dependent manner. Phytochemical investigation reveals that the fresh and the dry extract of RTV are rich in flavonoid, flavonol, phenols, tannin and proanthocyanidin, while the essential oil contained dimethy disulfide, dimethyl trisulfide, (methyl methylthio) methyl, 2,4-dithiapentane (11.35 percent) and (methylthio) acetic acid, 2- (methylthio) ethanol, 3-(methylthio) - and propanenitrile (7.20 percent). The fresh extract had higher radicals scavenging activity than the essential oil or dried extract, with 50 percentage inhibition of DPPH, hydrogen peroxide and lipid peroxidation at a concentration of 35.0 ± 0.12, 19.3 ± 0.11 and 17.9 ± 0.15 μg/ml respectively. Oral administration of methanolic extract of RTV in 125, 250 and 500 mg/kg to female Wistar rats significantly inhibited reduction of glutathione (GSH), superoxide dismutase (SOD) and catalase (CAT). The extracts also inhibited (p< 0.05) lipid peroxidation in normal, high cholesterol and diet induced atherosclerosis fed rats in a dose dependant manner. Also the extract (250 and 500 mg/kg/bwt/day) caused a significant (p<0.05) improvement in body weight of treated animals compared with untreated hypercholesterolemia control rats. The extracts also protected significantly (p<0.05) against atherogenic diet induced liver damage or fatty streaks formation in the aorta as revealed by histological examination. The anti-cholesterolemia and anti-atherosclerotic activities of the extract compared favorably well with standard drugs Gemfibrozil and Atorvastatin respectively. Conclusively, rhizomes of T. violacea possess significant anti-atherogenic activity and its mechanism of action(s) may be due to its antioxidant and anti-hypercholesterolemia properties. The results of this study also suggested that rhizome of T. violacea is relatively safe for human consumption and it may be used as an alternative to garlic.

Background. As a result of the AIDS pandemic, many people areimmuno compromised andopportunistic fungal infections (OFIs) such as candidiasis are common. Despite the widespread use of medicinal plants in South Africa, there is a dearth of knowledge regarding the use of such plants in the management of these infections. This study evaluates three South African medicinal plants (Arctotis arctotoides, Pittosporum viridiflorum, and Gasteria bicolor) traditionally used in the treatment of OFIs in HIV/AIDS patients, in the Eastern Cape Province, South Africa. Materials and methods. A six-stage process of documentation, evaluation and analysis of results was conducted: (1) Selection of medicinal plants most frequently used in the treatment of OFIs through ethnomedical studies and the survey of specialised literature; (2) Collection and preparation of the extract of each plant; (3) Antifungal evaluation of the crude plant extracts. (4) Phytochemical and antioxidant evaluation of the active crude plant extracts; (5) Cytotoxicity evaluation of the bioactive extracts using the Chang liver cell line, and (6) Statistical analysis of the results. Ethnobotanical information was obtained through interviews with traditional healers and AIDS patients with the aid of semi-structured questionnaires, direct observations and by reviewing studies reported in the literature. Following the approval from the University of Fort Hare‘s Ethics Committee, 101 HIV/AIDS patients were recruited through convenience sampling into an anonymous cross-sectional questionnaire study. The agar diffusion and micro-dilution methods were used to determine the antifungal activities of the hexane, acetone and aqueous extracts of A. arctotoides, G. bicolor and P. viridiflorum against 10 opportunistic fungi.

Diabetes mellitus is a chronic physiological carbohydrate metabolic disorder with significant impact on the economy, quality of life and life expectancy in South Africa. Herbal medicine has become the alternative therapy in the management of this disease. However, their safety and effectiveness have not been investigated. To address this, one of the plants used in Eastern Cape Province, South Africa, Bulbine abyssinica A. Rich (Asphodelaceae), was evaluated. Bulbine abyssinica is one of the species used in the management of diabetes mellitus. This plant was mentioned during an ethnobotanical survey conducted in Nkonkobe municipality of the Eastern Cape Province. Though a decoction prepared from the whole plant is used in the treatment of diabetes mellitus, the mechanism of action and its safety has not been elucidated. Thus, this research work was designed to contribute to the understanding of the possible mechanism of action of B. abyssinica as an antidiabetic medicinal plant and its toxic potentials to the users. The aqueous extract exhibited remarkable inhibitory activity onα-amylase (estimated inhibitory concentration (IC)50 value of 3.28 μg/ml), while the acetone extract exhibited weak inhibitory activity. The acetone extract exhibited notable α-glucosidase inhibitory activity (IC50 value of 4.27 μg/ml) while aqueous extract had significantly weak activity. The Lineweaver-Burk double reciprocal plots revealed that the aqueous extract exerts noncompetitive inhibition on the α-amylase activity while the acetone extract exerts a near competitive inhibitory pattern on the α-glucosidase activity. The extracts from the plant possessed high free radical scavenging activities, with acetone extract exhibiting the highest activities in all assay models used except with ferric reducing power and nitric oxide (NO) scavenging ability. The aqueous extract exhibited the highest ferric reducing power and nitric oxide radical mopping strength while the essential oil exhibited the highest scavenging activities with 1,1-diphenyl-2-picrylhydrazyl (DPPH) and relatively high ferric reducing power and nitric oxide scavenging ability. The acetone extract and the essential oil of this species exhibited higher albumin denaturation inhibition than the aqueous extract while the latter showed the greatest membrane lysis protection. The essential oil, acetone and aqueous extracts from this plant significantly inhibited the growth of Shigelle flexneri, Pseudomonas aeruginosa, Staphylococcus aureus and Enterococcus faecalis. Klebsiella pneumonia, Proteus vulgaris and Streptococcus pyogens growth were inhibited by acetone and aqueous extracts. The essential oil also showed inhibitory activity against Proteus vulgaris. However, the extracts were active against the growth of only three fungi species. The essential oil showed significant inhibitory activity against Trichophyton rubrum. The aqueous extract inhibited the growth of Microsporum gypseum while the acetone extract was active against Microsporum canis, and Microsporum gypseum. The carbohydrate, crude fibre, moisture, ash, crude protein and crude fat of approximately 74.8 percent, 8.9 percent, 8.8 percent, 8 percent, 7.7 percent and 0.6 percent, respectively, were detected in this plant. The species is characterized by moderate levels of oxalates, phytic acids, Vitamin A, Vitamin C and Vitamin E. Potassium and calcium were present in highest levels, while magnesium, iron, sodium, aluminium and phosphorus were moderately present. Manganese, zinc and copper where in low amounts. These vitamins and mineral elements were within their recommended daily allowance (RDA) in humans. The investigation also revealed appreciable amounts of total phenols, flavonoids, flavanols, proanthocyanidins and alkaloids in both acetone and aqueous extracts while saponins and tannins were in trace amounts. The essential oil was characterized by large quantities of terpenes (91.9 percent) and small fraction of esters (8.01 percent).

Objetivos: descrever a utilização de plantas medicinais e fitoterápicos concomitantemente com outros medicamentos na população do município de Teutônia, RS. Métodos: a pesquisa seguiu um modelo transversal descritivo e o instrumento de coleta de dados foi um questionário semi-estruturado. As entrevistas ocorreram no período de dezembro de 2005 a maio de 2006. As plantas coletadas foram identificadas botanicamente pelo Herbário ICN/UFRGS, do Departamento de Botânica da UFRGS. Resultados e Conclusões: Dos 196 entrevistados 87,2% eram do sexo feminino, com idade média de 44,4+13,86 anos. O uso de plantas medicinais foi relatado por 92,9% dos entrevistados, com média de 4,8+3,7 plantas por pessoa. Para a maioria dos respondentes (74,8%) a informação sobre o uso das plantas medicinais está baseada no conhecimento tradicional, sendo usadas principalmente para tratar sintomas de condições relacionadas ao trato gastrintestinal, ao sistema respiratório ou simplesmente como bebida. 58,6% das plantas medicinais referidas são cultivadas nas moradias dos entrevistados. Quanto à utilização de medicamentos, 37,8% dos entrevistados relataram que utilizam medicamentos sempre, 31,1% quando não se sentem bem e 8,7% somente quando consultam o médico. Das pessoas entrevistadas, 86,2% relataram o consumo de algum tipo de medicamento no mês anterior à entrevista. Foram citados 467 medicamentos, perfazendo a média de 2,4 medicamentos por pessoa. O uso de fitoterápicos foi de apenas 7,28%. A presença de doenças crônicas foi relatada por 45,9% do total dos entrevistados, correspondendo a 90 entrevistados. A hipertensão arterial foi a mais prevalente. Neste estrato, 90% utilizam plantas medicinais com média de 5,26+4,02 plantas por pessoa. A análise de comparação entre médias demonstrou que não existe diferença significativa entre o número médio de plantas medicinais utilizadas por pessoas com doenças crônicas e pessoas sadias (pcalc= 0,1056; p>0,05; t-Student). Os resultados deste estudo reforçam que o intercâmbio entre os conhecimentos científico e popular é um instrumento importante para o uso racional das plantas medicinais. / Objective: Describing the use of the medicinal plants and herbal medicines with other medicines in the population of Teutonia city, RS. Methods: The research followed a descriptive cross-sectional study model, using a questionnaire semi-structured in a data collection. The interviews occurred from December 2005 to May 2006. The collected plants were botanically identified by the herbal ICN/UFRGS, from Botanic department of UFRGS. Results and Conclusions: 196 were interviewed and among them 87.2% were female, at the average age of 44.4 around 13.86 years. The use of medicinal plants was told for 92.9 % of the interviewed, the average of 4.8, around 3.7 plants for each person. For the most of them (74.8%) the information about the use of medicinal plants is based in the traditional knowledge, being used mainly for treating the symptoms of conditions related to the gastrointestinal treat, to respiratory system or just as a beverage. In 58.6% medicinal plants mentioned, they are grown in the homes of the interviewed. About the using of the medicines, 37.8% of the interviewed tell that they always use the medicines, 31.1% use them when they do not feel well and 8.7 only when see the doctor. From the interviewed people, 86.2% told about the using of some kind of medicine in the month before the interview. Were mentioned 467 medicines, making the average of 2.4 medicines per person. The herbal medicines use was only of 7.28%. The presence of chronic diseases was related for 45.9% from the amount of interviewed, being 90 interviewed people. The arterial hypertension was the most prevalent. In this sample, 90% use medicinal plants with the average of 5.26 around 4.02 plants for each person. The analysis of comparison between averages showed that there is not significant differences between the middle number of medicinal plants used by people with chronic diseases and health people (p=0.1056; p>0.05; t-Student Test).The results of this study reinforces that the exchange between the popular and scientific knowledge is an important instrument for the rational use of the medicinal plants.

The aim of this Thesis was find out if adult population has experience with the use of medicinal plants, if people prefer local herbs to exotic plants, what is their knowledge in the preparation of herbal products and if they ever know what the herbs are and what is the meaning of the term Phytotherapy. Another aim was the creation of electronic manual of the most used herbs, which would include information on sellected medicinal plants and their effects on human body. In this thesis there are comprehensively summarized the theroetical and practical knowledge, which should be beneficial for general population. Research survey in the form of a questionnaire was focused on experience and utilization of herbal therapy in adult population. That was realized in South Bohemian and Pilsen Regions. Results of this work point out, that respondents still consider herbal medicine as an important supplement to classical medical treatment. On the basis of the research there were conclusion written and made the recommendation for practise.

Submitted by Ana Hilda Fonseca (anahilda@ufba.br) on 2013-04-09T15:57:22Z No. of bitstreams: 1 Dissertação Alessandra (4).pdf: 2685859 bytes, checksum: 5365f2b719ecbda7464c4a77b11585ec (MD5) / Approved for entry into archive by Ana Hilda Fonseca(anahilda@ufba.br) on 2013-04-18T17:37:07Z (GMT) No. of bitstreams: 1 Dissertação Alessandra (4).pdf: 2685859 bytes, checksum: 5365f2b719ecbda7464c4a77b11585ec (MD5) / Made available in DSpace on 2013-04-18T17:37:08Z (GMT). No. of bitstreams: 1 Dissertação Alessandra (4).pdf: 2685859 bytes, checksum: 5365f2b719ecbda7464c4a77b11585ec (MD5) Previous issue date: 2009 / CAPES / O presente trabalho é uma contribuição à pesquisa de plantas medicinais, e descreve o estudo farmacognóstico das espécies Averrhoa bilimbi e Poiretia bahiana no que diz respeito aos aspectos botânicos, fitoquímicos e à avaliação de atividade biológica. As folhas e frutos da espécie A. bilimbi foram caracterizados pelos aspectos morfoanatômicos, e apenas de seus frutos foram realizadas extrações com metanol, seguido de fracionamento para obtenção dos extratos brutos hexânico, clorofórmio e acetato de etlia. Estes extratos foram avaliados quanto ao potencial citotóxico através do bioensaio com o microcrustáceo Artemia salina. Os extratos brutos clorofórmio e acetato de etila foram submetidos ao fracionamento e isolamento de substâncias através de técnicas cromatográficas. A identificação das substâncias isoladas foi realizada através de métodos espectroscópicos, tais como RMN de 1H e 13C. Para a espécie P. bahiana, o estudo envolveu a caracterização anatômica de suas folhas e obtenção de extratos brutos hexânico, clorofórmio, acetato de etlia e hidroalcoólico através de técnicas cromatográficas. Os extratos obtidos foram avaliados quanto ao potencial citotóxico frente a A. salina, e a capacidade seqüestradora de radical livre, medida pelo método DPPH (2,2-difenilpicril- hidrazil). Os resultados apresentados no estudo farmacobotânico das espécies A. bilimbi e P. bahiana permitiram levantar características muito peculiares nos aspectos morfoanatômicos. A espécie A. bilimbi apresentou um elevado potencial citotóxico, evidenciado pela morte de 100% dos microcrustáceos em todos os extratos avaliados. A partir do fracionamento e isolamento das substâncias presentes nos extratos acetato de etila e clorofórmio da espécie A. bilimbi foram isoladas as substâncias 3β-O-β-D-glicopiranosil sitosterol e 3,5-dimetoxi-benzaldeído, até então não descritas para a espécie. Já a espécie P. bahiana apresentou apenas citotoxicidade nos extratos hexânico e clorofórmio, entretanto foi observada atividade antioxidante dos extratos hidroalcoólico, clorofórmio e acetato de etila através do teste com o DPPH, demonstrando ter potencial como fonte de substâncias antioxidantes. / Salvador

Plants have been used as sources of medicinal agents for millennia. Plant extracts and derived active principles have served as a major source for new pharmaceuticals for treatment of malignant tumors. In this work, we described the plants most used as anticancer agents, by the population treated on the Oncology Service of the University Hospital (Maceió-AL). The aim of this work was evaluate the toxicological effects of those plants and to value the antitumoral activity of the extracts and juices against human tumor cell lines and in neoplasic assays in vivo. Initially, the plants were collected, identified, and the hydroethanolic extracts and juices of Aloe vera and Euphorbia tirucalli were prepared. In the pharmacological approach, the follow assays were done: toxicity against brine shrimp Artemia salina, in vitro test against human tumor cell lines and in vivo evaluation using sarcoma 180 in mice. The plants were Annona muricata, Aloe vera, Schinus terebenthifolius, Hyptis pectinata, Stryphnodendron adstringens, Euphorbia tirucalli, Caesalpinia bonduc and Cnidosculus urens. The phytochemical screening showed the presence of flavonoids, tanins, saponins and sterols. The Artemia salina essay suggested moderate toxicity of the Stryphnodendron adstringens, Annona muricata e Euphorbia tirucalli. Hyptis pectinata was considered atoxic. In the preliminary in vitro assay only two species showed moderate cytotoxicity: Annona muricata (82%, 86% and 71% of inhibition) against SF295, HCT-8 and MDA-MB435 line cells, respectively and Hyptis pectinata (62% of inhibition) against HCT-8.line cells. Both plants presented no hemolytic effect and in the evaluation of the cytotoxicity activities against peripheral blood mononuclear cells, Annona muricata showed the biggest activity, although had shown bigger toxicity than Hyptis pectinata to PBMC. In the in vivo tests, Hyptis pectinata, Schinus terebinthifolius, Annona muricata and Euphorbia tirucalli inhibited the sarcoma 180 (70,5% - 67,1% - 58,9% and 57,5%). The histopathological analysis of the liver, kidneys and spleen of animals which was treated showed absence of lesions. Take together these results suggest that only two plants (Hyptis pectinata and Annona muricata) used by the population showed potential anti-tumor activity, at least in the used pharmacological models in vitro and in vivo. / Fundação de Amparo a Pesquisa do Estado de Alagoas / Plantas têm sido usadas como fonte de medicamentos há milênios. Extratos de plantas e princípios derivados ativos têm servido como a maior fonte de novos fármacos para tratamento de tumores malignos. Neste trabalho, descrevemos as plantas mais utilizadas como agentes anticancerígenos, pela população tratada no Centro de Oncologia do Hospital Universitário/UFAL (Maceió-AL). O objetivo deste trabalho foi avaliar os efeitos toxicológicos destas plantas e analisar a ação de seus extratos e sucos sobre células tumorais em modelos in vitro e em neoplasias in vivo. Inicialmente, as plantas foram coletadas, identificadas, e os extratos hidroetanólicos e sucos de Aloe vera e Euphorbia tirucalli foram preparados. Na investigação farmacológica, os seguintes ensaios foram feitos: Toxicidade frente ao microcrustáceo Artemia salina, teste in vitro contra células de linhagem tumoral humana e avaliação in vivo utilizando Sarcoma 180 em camundongos. As plantas foram: Annona muricata, Aloe vera, Schinus terebenthifolius, Hyptis pectinata, Stryphnodendron adstringens, Euphorbia tirucalli, Caesalpinia bonduc e Cnidosculus urens. O screening fitoquímico mostrou a presença de flavonóides, taninos, saponinas e esteróides. O ensaio de Artemia salina sugeriu alta toxicidade de Stryphnodendron adstringens, Annona muricata e Euphorbia tirucalli; Hyptis pectinata foi considerada atóxica. No ensaio preliminar in vitro, apenas duas espécies exibiram moderada citoxicidade: Annona muricata (82%, 86% e 71% de inibição) contra as linhagens celulares SF295, HCT-8 e MDA-MB435, respectivamente e Hyptis pectinata (62% de inibição) contra a linhagem celular HCT-8. Ambas as plantas apresentaram nenhum efeito hemolítico e na avaliação da atividade citotóxica em células mononucleares do sangue periférico (PBMC), Annona muricata exibiu maior atividade, embora tenha apresentado maior toxicidade que Hyptis pectinata.para PBMC. Nos testes in vivo, Hyptis pectinata, Schinus terebinthifolius, Euphorbia tirucalli e Annona muricata e inibiram o Sarcoma 180 (70,5% - 67,1% - 58,9% e 57,5%). A análise histopatológica de fígado, rins e baço dos animais tratados, informou ausência de lesões. Em conjunto estes resultados sugerem que apenas duas plantas (Hyptis pectinata e Annona muricata) usadas pela população mostraram potencial atividade antitumoral, pelo menos nos modelos farmacológicos in vitro e in vivo utilizados.

Brazil is considered a holder country of a great biodiversity, which the popular use of medicinal plants contributes an important source to discovery the new active compounds. Plants of the family Bignoniaceae and Tabebuia genus are widely used in traditional medicine in several countries for the treatment of various diseases and have an excellent potential for scientific studies, which may result in the discovery of new compounds having therapeutic interest. In this context, evidence the need to assess the biological potential of Tabebuia aurea, investigate their cytotoxic, antimicrobial activity, antiradical, antiedematogenic and lead the phytochemical screening to identify classes of bioactive compounds. We evaluated two ethanolic extracts from flowers and leaves, both subject to cell viability assay Metiltetrazólio by the colorimetric method for assessing the presence of cytotoxicity. The antimicrobial activity was evaluated using the broth microdilution assay, which provided the minimum inhibitory concentration (MIC) of the extracts against microbial growth. We performed the test ear edema induced by capsaicin to determine the antiedematogenic action and the evaluation of antiradical activity, the test verifies that the scavenging capacity of free radical opposite to 2,2-difenil-1-picrilidazila (DPPH) radical. The cell viability assay showed that the leaves ethanolic extract showed no cytotoxicity, since the LC50 value was greater than the maximum concentration used (≥ 1,0 mg ml-1), while the flowers ethanolic extract had no cytotoxicity at concentrations ≤ 0,5 mg mL-1. The ethanol extracts of flowers and leaves of T. aurea showed antibacterial activity, especially toward those Gram-positive bacteria. The flowers ethanol extract proved to be active with bactericidal action against the strain of S. epidermidis at MIC of 0,06 mg mL-1 and the leaves ethanol extract moderately active against S. epidermidis (CIM: 0,25 mg ml-1) and S. aureus (MIC: 0,50 mg mL-1) with bacteriostatic action to both strains. Both extracts inhibited edema formation induced by capsaicin. Treatment with flowers ethanol extract showed a significant inhibition of edema of 40,50% and the ethanol extract of leaves of 41,73%. The ethanol extracts of T. aurea showed no antiradical activity against the DPPH radical. The plant species used was identified by the presence of organic acids, naphthoquinones, anthraquinones and saponins. These results scientifically prove the antiedematogenic potential of the species and its promising antibacterial activity, with no cytotoxicity. Representing security evidence on the therapeutic use of plant species in preclinical in vivo tests. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O Brasil é considerado um país detentor de uma grande biodiversidade, no qual o uso popular de plantas medicinais contribui como importante fonte para a descoberta de novos compostos ativos. As plantas da família Bignoniaceae e do gênero Tabebuia são amplamente utilizadas na medicina tradicional de vários países, para o tratamento de diversas doenças e possuem um excelente potencial para estudos científicos, que podem resultar na descoberta de novos compostos com interesse terapêutico. Nesse contexto, evidenciou-se a necessidade de avaliar o potencial biológico de Tabebuia aurea, investigando sua atividade citotóxica, antimicrobiana, antirradicalar, antiedematogênica e conduzindo a prospecção fitoquímica para identificar as classes de compostos bioativos. Foram avaliados dois extratos etanólicos, um proveniente de flores e outro de folhas, ambos submetidos ao ensaio de viabilidade celular pelo método colorimétrico de Metiltetrazólio, para avaliar a presença de citotoxicidade. O potencial antimicrobiano foi avaliado com o teste de microdiluição em caldo, o qual forneceu a concentração inibitória mínima (CIM) dos extratos frente ao crescimento microbiano. Para determinar a ação antiedematogênica, realizou-se o teste de edema de orelha induzido por capsaicina e para a avaliação da atividade antirradicalar, o ensaio que verifica a capacidade sequestradora de radicais livres frente ao radical 2,2-difenil- 1-picrilidazila (DPPH). O ensaio de viabilidade celular revelou que o extrato etanólico das folhas não evidenciou citotoxidade, uma vez que a CL50 foi superior a concentração máxima utilizada (≥ 1,0 mg mL-1), enquanto que o extrato etanólico de flores não teve citotoxidade nas concentrações ≤ 0,5 mg mL-1. Os extratos etanólicos de flores e de folhas de T. aurea apresentaram atividade antibacteriana, principalmente, frente às bactérias Gram-positivas. O extrato etanólico de flores demonstrou-se ativo, com ação bactericida, frente à linhagem de S. epidermidis na CIM de 0,06 mg mL-1 e o extrato etanólico de folhas moderadamente ativo frente a S. epidermidis (CIM: 0,25 mg mL-1) e S. aureus (CIM: 0,50 mg mL-1), com ação bacteriostática para ambas as linhagens. Ambos os extratos inibiram a formação do edema induzido por capsaicina. O tratamento com o extrato etanólico de flores apresentou uma porcentagem de inibição do edema de 40,50% e o extrato etanólico de folhas de 41,73%. Os extratos etanólicos de T. aurea não apresentaram atividade antirradicalar frente ao radical DPPH. Foram idenficados na espécie vegetal avaliada a presença de ácidos orgânicos, naftoquinonas, antraquinonas e saponinas. Esses resultados comprovam cientificamente o potencial antiedematogênico da espécie e sua promissora atividade antibacteriana, com ausência de citotoxidade. Representando indícios de segurança na utilização terapêutica da espécie vegetal nos testes pré-clínicos in vivo.

Clusia nemorosa is distributed throughout Brazil and its extract is widely used in popular medicine to treat different diseases. Although the popular use of extract of C. nemorosa, the scientific reported on their pharmacological and phytochemical analysis are scarce. In the present study, we aimed to evaluate the antinociceptive and antiinflammatory effect of hexane extract of Clusia nemorosa-derived stem bark (EHC). To evaluate the antinociceptive effect of EHC classical models of the study of pain were used, such as the abdominal writhing test induced by acetic acid, formalin test and hot plate test. In order to evaluate anti-inflammatory effect of EHC, the model of paw edema induced by carrageenan were used. Rota-rod testing and toxicological analysis in vivo and in vitro were performed. Mices submitted to abdominal writhing test, was observed a significant reduction in nociceptive response induced by acetic acid, and these effects persisted for up to 2 h after treatment. Moreover, it is important emphasizing that the EHC did not induce changes in motor function of animals when the rota-rod test was performed. In order to investigate the possible mechanism of action of EHC, the animals were pretreated with naloxone (5mg/kg), yohimbine (1 mg/kg), atropine (5 mg/kg), metoclopramide (1 mg/kg), haloperidol (2 mg/kg) and L -NAME (20 mg/kg). Only when used metoclopramide, a serotoninergic and dopaminergic antagonist receptor, it was observed an inhibition of the antinociceptivo effect of EHC, suggesting that the serotoninergic pathway may be involved in the action of the EHC, as haloperidol, a dopaminergic antagonist did not reverse the antinociception of EHC. In addition, EHC did not induce changes in the nociceptive response of animals when assessed by the hot plate test, discarding the action in the central nervous system. Then, when the animals were treated with EHC and submitted to the formalin test, it was observed that only the second phase of this test was inhibited by the EHC, indicating inhibitory actions on pain of EHC from inflammatory origin. To evaluate possible toxic effects of EHC, animals were treated for 7 consecutive days. Then after treatment, were not observed changes in behavior, blood cellularity and cytotoxicity in vitro, which ruled out possible of toxic effects in these preliminary tests. Through analysis of gas chromatography coupled with mass spectrum, the major component present in the extract was identified, friedelin, who presented antinociceptive effects on pain and inflammation of neurogenic origin, as evidenced by the formalin test. Although this model of pain induced by formalin, it was observed that the antinociceptive effects friedelin were maintained when the EHC was administered orally. In addition, friedelin inhibited the paw edema induced by carrageenan. Lastly, in vitro assays, this triterpene did not affect cell viability as measured by MTT test. Together, our results support the popular use of this plant based on their anti-inflammatory and antinociceptive effects. In addition, we report the first time the presence of pentacyclic triterpenoid, friedelin on hexane extract of Clusia nemorosa-derived stem bark, triterpenoid which is possibly responsible for the pharmacological effects evaluated. / Conselho Nacional de Desenvolvimento Científico e Tecnológico / A espécie Clusia nemorosa é bem distribuída pelo território brasileiro e seus extratos são amplamente utilizados pela medicina popular para tratar diferentes doenças. Mesmo sendo comum o consumo dos extratos de C. nemorosa (EHC), os registros científicos sobre as propriedades farmacológicas e análises fitoquímicas desta espécie ainda mostram-se escassos. Assim, no presente estudo objetivamos avaliar o efeito antinociceptivo e anti-inflamatório do extrato hexânico obtido a partir das cascas do caule de Clusia nemorosa. Para traçar o perfil antinociceptivo foram utilizados os modelos de contorção abdominal induzido por ácido acético, formalina e o teste de placa quente. Com propósito de avaliar o perfil anti-inflamatório utilizamos o teste de edema de pata induzido por carragenina. Para melhor compreender as ações do EHC, submetemos os animais tratados com o extrato ao teste de rota-rod e a análises toxicológicas in vivo e in vitro. No ensaio de contorção abdominal foi observada uma redução na resposta nociceptiva induzida por ácido acético, sendo estes efeitos persistentes por até 2 h após tratamento com EHC. Através do teste de rota rod, verificamos que o EHC não induziu alterações na motricidade dos animais. Com propósito de investigar o possível mecanismo de ação deste extrato os animais foram pré-tratados com naloxona (5 mg/kg), ioimbina (1 mg/kg), atropina (5 mg/kg), metoclopramida (1 mg/kg), haloperidol (2 mg/kg) e LNAME (20 mg/kg). Apenas quando utilizado a metoclopramida, antagonista de receptores serotoninérgicos e dopaminérgicos, ocorreu inibição do efeito antinociceptivo do EHC, sugerindo que as vias serotoninérgicas possam estar envolvidas nas ações do extrato, pois o haloperidol, um antagonista dopaminérgico não reverteu a antinocicepção do EHC. Em adição, o tratamento com EHC não induziu alterações na resposta nociceptiva dos animais quando avaliados pelo teste da placa quente, descartando possíveis ações no Sistema Nervoso Central. Em seguida, quando os animais tratados com extrato foram submetidos ao teste de formalina, foi observado que apenas a segunda fase deste teste foi inibida, indicando ações inibitórias sobre a dor de origem inflamatória, revertida pelo pré-tratamento com metoclopramida. Quando os animais foram submetidos ao tratamento por 7 dias consecutivos com o EHC não foram observadas alterações no comportamento, na celularidade do sangue e citotoxicidade in vitro, o que descartou possíveis efeitos tóxicos nestes testes preliminares. Através de análise de cromatografia gasosa acoplada a espectro de massa, foi identificado o componente majoritário presente no extrato, a friedelina (1 e 10 mg/kg; i.p.), que apresentou efeitos atinociceptivos na dor de origem neurogênica e inflamatória, evidenciado pelo teste de formalina. Ainda neste modelo, foi observado que a friedelina manteve seus efeitos antinociceptivos na dor de origem inflamatória quando administrado por via oral. Além disso, a friedelina inibiu o edema de pata induzido por carragenina. Em adição, em ensaios in vitro, este triterpeno não alterou a viabilidade celular. Em conjunto, nossos resultados sustentam o uso popular desta planta tendo por base seus efeitos antinociceptivos e anti-inflamatórios. Além disso, relatamos pela primeira vez a presença do triterpenóide pentacíclico, friedelina, na casca de C. nemorosa, sendo este triterpenóide possivelmente responsável pelos efeitos farmacológicos avaliados.

The diploma thesis concentrates on medicinal plants, their use and effects in the treatment of diseases. The thesis summarizes some effects of medicinal substances contained in plants and herbs. They are suitable for use in selected diseases. The research part of diploma thesis deals with the mapping of methods of treatment and production of herbal products among university students. The thesis also examines whether students are inclined to use more herbal therapy over conventional pharmaceutical drugs. It investigated whether information on herbs in schools sufficient. KEY WORDS medicinal plants, plant's medicinal substances, herbs, products, phytotherapy