Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The aim of this study was to evaluate the effect of the essential oil of Lippia alba (EOLA) as a feed additive on ionoregulatory and metabolic parameters, and pituitary hormones expression in silver catfish, Rhamdia quelen. Fish were fed for 20 days with different concentrations of EOLA (0.0 control, 0.25 and 0.50 mL kg-1 food). Plasma Na+, Cl- , K+ and cortisol, and metabolic parameters were not affected by the diet, with the exception of ALT (alanine aminotransferase), which was higher in the liver of fish fed 0.50 mL EOLA kg-1 food. Fish fed 0.25 mL EOLA kg-1 food presented higher Na+/K+-ATPase activity and somatolactin expression, but H+-ATPase activity and growth hormone and prolactin expression did not change. The EOLA can be used as a dietary supplement for silver catfish at the evaluated concentrations, but using 0.25 mL EOLA kg-1 food seems to be more suitable than 0.50 mL EOLA kg-1 food since the latter may be related to liver damage. / O objetivo deste estudo foi analisar o efeito do óleo essencial de Lippia alba (EOLA) adicionado à ração de Rhamdia quelen (jundiá) em parâmetros metabólicos, osmorregulatórios e endócrinos. Os peixes foram alimentados por 20 dias com diferentes concentrações de OELA (0,00; 0,25 e 0,50 mL kg-1 ração). Os peixes foram alimentados durante 20 dias com diferentes concentrações de OELA (0,0 - controle, 0,25 e 0,50 mL kg-1 de ração). Parâmetros metabólicos, Na+, Cl-, K+ e cortisol no plasmas não foram afetados pela dieta, com a exceção de ALT (alanina aminotransferase), que foi maior no fígado dos peixes alimentados com 0,50 mL OELA kg -1 de ração. Os peixes alimentados com 0,25 mL EOLA kg-1 de ração, apresentaram maior atividade de Na+/K+-ATPase e da expressão da somatolactina, mas a atividade de H+-ATPase e da expressão do hormônio do crescimento e da prolactina não alterou-se. O OELA pode ser utilizado como um suplemento dietético para o jundiá nas concentrações testadas, mas 0,25 mL EOLA kg -1 de ração, parece ser melhor do que a concentração de 0,50 mL kg-1, uma vez que esta última pode estar relacionada com danos no fígado.

Magister Scientiae - MSc / This investigation focused on Rhoicissus tomentosa, belonging to the family, Vitaceae in an attempt to assess the phytochemistry of this plant which is widely used by traditional healers in South Africa to ensure the safe delivery during pregnancy and childbirth (Hutchings et al., 1996). / South Africa

Kalaharituber pfeilii (Henn.) Trappe & Kagan-Zur commonly known as the “Kalahari truffle” is a desert truffle species identified from the Kalahari region of southern Africa. Two other species, Eremiomyces echinulatus (Trappe & Marasas) Trappe & Kagan-Zur and Mattirolomyces austroafricanus (Trappe & Marasas) Trappe & Kovacs are also known to occur in other parts of southern Africa. Truffles are hypogeous fruiting bodies of Ascomycetes, important to humans for their nutritional value and medicinal characteristics. These truffles are known as desert truffles as they prefer to occur under arid or semi-arid conditions characteristic of deserts. Truffle development depends on the presence of a mycorrhizal host, associated microorganisms as well as soil and climatic characteristics. It has been suggested that K. pfeilii has a suspected broad plant host range which includes herbaceous to woody trees and shrubs. However, these relationships have not been verified. Indigenous people of the Kalahari believe that truffles are found under grasses. In the Kalahari, truffle fruiting bodies are often found entangled in Stipagrostis ciliata (Desf.) De Winter var. capensis (Trin. & Rupr.) De Winter roots. S. ciliata, also known as the tall bushman-grass, is the most common grass found in the Kalahari. The objective of this study was to provide conclusive evidence that S. ciliata var. capensis is a host of the Kalahari truffle. Truffle fruiting bodies and grass roots from where the truffles were found were collected from Upington, South Africa. The fruiting bodies were identified by observing their morphological characteristics using the ‘Keys of Truffle genera’. All observed physical properties were similar to those of K. pfeilii and further identification was done using molecular techniques. DNA was extracted from the fruiting bodies, mycelial cultures, rhizosheaths and from the S. ciliata var. capensis grass roots, which were then amplified using the specific K. pfeilii specific primers TPF3 and TPR1 and sequenced. The obtained sequence results confirmed that the collected fruiting bodies were those of the K. pfeilii and the molecular techniques also confirmed that K. pfeilii DNA was present in the S. ciliata var. capensis rhizosheath and root cells. Microscopy showed an ectendomycorrhizal association between K. pfeilii and S. ciliata var. capensis. Mycorrhizal resynthesis experiments were conducted to establish this mycorrhizal association in-vitro. They were unsuccessful because of the structure of the grass and the availability of contaminants. And more...

Pour la première fois, l’étude de la flore fongique endophytique de cinq plantes médicinales soudanaises : Calotropis procera (Ait.), Catharanthus roseus (L.), Euphorbia prostrata (Ait.), Trigonella foenum-graecum (L.), and Vernonia amygdalina (Del.) a été réalisée. Un total de 23 souches de champignons endophytes ont été isolées à partir des plantes après la stérilisation de surface puis les différentes analyses biologiques ont été effectuées. Les extraits bruts d’acétate d’éthyle de 21 endophytes ainsi que de leurs plantes hôtes ont été évalués pour leur teneur en phénols totaux et leur activité antioxydante en utilisant respectivement la méthode colorimértrique Folin-Ciocalteu et le piégeage des radicaux libres par la méthode 1,1,-diphényl-2-picrylhydrazil (DPPH) in vitro. Une évaluation générale de la cytotoxicité de 16 endophytes sélectionnés ainsi que de leurs plantes hôtes a été réalisée selon le test MTT sur trois types de cellules cancéreuses : carcinome du sein humain (MCF7), adénocarcinome du côlon (HT29 et HCT116). Ces extraits ont été aussi testés, selon la méthode de dilution en bouillon, sur deux souches bactériennes représentatives, Escherichia coli et la souche résistante à la méthicilline de Staphylococcus aureus. La teneur en phénols totaux (89,9 ±7,1 mg Equivalent d’Acide Gallique EAG/g) ainsi que l’activité antioxydante (IC50: 18±0,1 µg/mL) les plus élevées ont été observées pour l’endophyte, Aspergillus terreus 2 isolé à partir des graines de T. foenum-graecum. Byssochlamys spectabilis a montré l’activité cytotoxique la plus importante (1,51 ± 0,2 µg/mL), suivi par Cladosporium cladosporioides 2 (10,5 ± 1,5 µg/mL), puis par Alternaria sp. (13,5 ± 1,8 µg/mL). Seules six souches ont montré une activité contre S. aureus avec des valeurs de MIC qui se situent entre 0,125 et 2 mg/mL dont: Alternaria alternata (0,125 mg/mL), Alternaria sp. (0,250 mg/mL), Byssochlamys spectabilis (0,5 mg/mL). 10 composés purs (0,3 à 40 mg) ont été isolés à partir des extraits bruts d’acétate d’éthyle de Curvularia papendorfii. Le nouveau composé pur (AFB) 3,7,11,15-Tetrahydroxy-18-hydroxymethyl-14,16,20,22,24-pentamethyl-hexacosa-4E,8E,12E,16,18-pentaenoic acid (acide Khartomique) a montré une activité antibactérienne modérée contre S. aureus avec une CIM de 62,5 µg/mL et une faible activité cytotoxique sur les cellules MCF7 avec une IC50 > 100 µM. Le composé pur AF1 a montré une activité cytotoxique modérée sur les cellules HT29 avec une IC50 de 29,78 µM et une très faible activité antibactérienne contre S. aureus. Ces deux composés ne présentent pas d’activité antioxydante. / This study investigated, for the first time, the endophytic fungi flora of five Sudanese medicinal plants: Calotropis procera (Ait.), Catharanthus roseus (L.), Euphorbia prostrata (Ait.), Trigonella foenum-graecum (L.) and Vernonia amygdalina (Del.). A total of 23 endophytic fungal strains were isolated from the plants after surface disinfection and different biological tests were performed. Total phenolic content (TPC) and total antioxidant activity of ethyl acetate crude extracts of 21 endophytes and their host plants were estimated using respectively the Folin-Ciocalteu colorimetric method and 1,1,-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging in vitro method. General evaluation of the cytotoxicity of 16 selected endophytes and their host plants was performed by the MTT assay using cancer cells type: Human breast carcinoma (MCF7) and Colon adenocarcinoma (HT29 and HCT116). Preliminary antibacterial screening was done for the 16 endophytes. These extracts were also tested against two representative bacterial strains, Escherichia coli and methicillin-resistant Staphylococcus aureus, by broth dilution tests. The endophyte, Aspergillus terreus 1 from T. foenum-graecum seeds had the highest TPC in term of Gallic Acid Equivalent (89.9 ± 7.1 mg GAE/g) and antioxidant activity (IC50: 18±0.1µg/mL). Byssochlamys spectabilis showed strong cytotoxicity (1.51 ± 0.2 µg/mL) followed by Cladosporium cladosporioides 2 (10.5 ± 1.5 µg/mL), then Alternaria sp. (13.5 ± 1.8 µg/mL). Only six strains showed activity against methicillin-resistant S. aureus with MIC values ranging between 0.125-2 mg/mL, Alternaria alternata (0.125 mg/mL) Alternaria sp. (0.250 mg/mL) and Byssochlamys spectabilis values (0.5 mg/mL). Ten pure compounds (0.3 to 40 mg) were isolated from ethyl acetate crude extract of Curvularia papendorfii .The new pure compound (AFB) 3,7,11,15-Tetrahydroxy-18-hydroxymethyl-14,16,20,22,24-pentamethyl-hexacosa-4E,8E,12E,16,18-pentaenoic acid (Khartoumic acid) revealed moderate antibacterial activity against S. aureus with MIC value 62.5 µg/mL and weak cytotoxicity with a IC50 > 100 µM against MCF7 cells. The pure compound AF1 showed moderate cytotoxic activity with IC50 value of 29.78 µM against HT29 and weak antibacterial activity with MIC 250 µg/mL against S. aureus. Both compounds displayed no antioxidant activity.

Over the past 25 years, there has been a resurgence of worldwide scientific research in the fields of ethnopharmacology. The Western world has acknowledged the continued use of traditional medicines by the majority of third world countries, and the need for novel drug development. Hence, much of the pharmaceutical research in recent years has focused on the ethnobotanical approach to drug discovery (Light et al., 2005). In South Africa, as in most developing parts of the world, traditional herbal medicine still forms the backbone of rural healthcare. The government health services in South Africa provide only western medical care although the majority of the population consult traditional healers for some or all of their healthcare needs (McGaw et al., 2005). Medicinal plants like Harpephyllum caffrum are used as blood purifiers or emetics (Watt and Breyer-Brandwijk, 1962), and also for treating acne and eczema. The antimicrobial activity of this plant can be used to treat septicaemia, which is ranked the sixth leading cause of death among neonates and the eighth leading cause of death for infants through the first year of life (Heron, 2007). In this study, the plants investigated for antimicrobial activity were Harpephyllum caffrum, Hermannia cuneifolia, Chironia baccifera, Rhigozum obovatum, Felicia muricata and Pentzia incana. These plants were tested against ATTC (American Type Culture Collection) strains and microorganisms isolated from clinical isolates of patients suffering from septicaemia. The assay methods used included the agar diffusion method using the Mast multipoint inoculator, the microtitre dilution method were used to determine the minimum inhibitory concentration, thin layer chromatography fingerprints accompanied by bioautographic assay were used to detect the inhibition of bacterial growth by active compounds separated from plant extracts and the Ames test was required to assess the possibility of bacterial mutagenesis upon the exposure to plant extracts which can lead to carcinogenicity. In agar diffusion method, extracts of Harpephyllum caffrum inhibited nine strains of Candida albicans, three species of Acinetobacter and four strains of E.faecalis. Extracts of Hermannia cuneifolia inhibited four strains of B.cereus and three strains of Staphylococcus aureus. Extracts of Chironia baccifera inhibited one strain of Acinetobacter and five strains of E.faecalis. Extracts of plants Rhigozum obovatum, Felicia muricata, and Pentzia incana showed no antimicrobial activity. In the microtitre dilution method used to determine the minimum inhibitory concentration (MIC), the results were different from the agar diffusion method. More activity was observed. Extracts of Harpephyllum caffrum inhibited three strains of E.coli, six strains of S.aureus, three species of Acinetobacter and one strain of Klebsiella pneumonia. Extracts of Hermannia cuneifolia inhibited four strains of B.cereus, three strains of S.aureus, two strains of K.oxytoca and one species of Acinetobacter. Extracts of Chironia baccifera inhibited three strains of S.aureus, one strain of MRSA, one species of Acinetobacter and one strain of S.haemolyticus. The MIC values ranged from 0.049 to 6.25mg/ml. Using the thin layer chromatography fingerprints, bioautography showed the presence of various inhibitory chemical compounds. Methanol and acetone extracts of Harpephyllum caffrum, separated very well and showed various inhibition zones on exposure to Candida albicans, Enterococcus faecalis and Staphylococcus aureus. The different inhibition zones were recorded as Rf In the Ames test (Maron and Ames, 1983) the methanol and acetone extracts of Harpephyllum caffrum and Hermannia cuneifolia were negative which means they were devoid of any mutagenic properties. Methanol extracts of Harpephyllum caffrum showed similar results in the Ames assay as reported by Verschaeve and Van Staden (2008). values ranging from 0.25 to 0.95. The zones indicate the different inhibiting chemical compounds present in the plant. Petroleum ether, ethyl acetate, chloroform and formic acid were the solvents used in the assay in the ratio 8:7:5:1, respectively. Establishing the antimicrobial activity of these plants contribute to the systematic scientific investigation of indigenous South African medicinal plants.

Alcohol abuse is a very common practice (just like in many other parts of the world) in Nkonkobe Municipality, Eastern Cape Province, South Africa. This is associated with liver disease. An ethnobotanical survey of plants used for the treatment of alcohol-induced liver damage in Nkonkobe Municipality was conducted. During the survey and also from information gathered in the literature, Pelargonium reniforme Curtis, was prominently mentioned, among other plants, as the species used generally for the treatment of alcohol-induced liver damage. This project was designed to evaluate the effects of the plant on alcohol-induced liver damage, including its antioxidant and antimicrobial properties. It also involves safety evaluation studies to determine if the plant is safe for consumption. Studies using rats of the Wistar strain were carried out to determine the protective and curative effects of P. reniforme on alcohol-induced liver damage. Results obtained showed that the plant extract can protect the liver cells as well as enhance recovery from tissue damage. The plant also showed good antimicrobial and antioxidant activity and this further validates its use in the treatment of liver diseases. Safety evaluation studies of the extract were carried out by investigating the effects of the oral administration on some haematological and biochemical parameters in male Wistar rats. The results obtained from the study suggest that the plant extract is not toxic at the doses used and is therefore safe for medicinal uses. The results of the various bioassays carried out in this project have justified the traditional uses of P. reniforme for the treatment of alcohol-induced liver damage.

Nos travaux de doctorat s’inscrivent dans la démarche de la valorisation de la biodiversité végétale cambodgienne. Les objectifs portaient sur les travaux phytochimique et pharmacologique de deux plantes cambodgiennes : Vernonia cinerea et Vernonia elliptica. Une méthode de dosage par CLHP du composé 8α-tigloyloxyhirsutinolide 13-O-acetate (H1) appartenant à la classe des lactones sesquiterpènes de V. cinerea a été mise au point et validée dans le but de déterminer la teneur de ce composé. La teneur en H1 varie entre 0,08 % à 0,17 % pour les échantillons étudiés provenant de 4 régions du Cambodge. L’évaluation de l’activité antiproliférative in vitro des extraits de V. cinerea, et du composé H1 a été réalisée vis-à-vis de deux lignées cellulaires cancéreuses humaines HT29 et HepG2. Le composé H1 présente une CI50 similaire à celle du contrôle positif 5FU, 4,3 ± 0,2 versus 5,4 ± 0,9 µM vis-à-vis de la lignée HepG2. Les résultats intéressants obtenus sur V. cinerea nous ont incités à travailler sur d’autres espèces du genre Vernonia qui poussent au Cambodge, Vernonia elliptica. L’étude phytochimique menée sur cette plante a permis d’isoler un lignane [(+)-Syringaresinol] et deux lactones sesquiterpènes [(+)-8,13-diacetyl-piptocarphol et Glaucolide B]. Ces composés sont décrits dans cette plante pour la première fois. L’étude in vitro de l’activité antiproliférative des extraits de V. elliptica montre que l’extrait dichlorométhane obtenu à partir de l’extrait aqueux est très actif. / Our PhD work is in the process of valorization of the Cambodian plant biodiversity. The objectives focused on phytochemical and pharmacological studies of two Cambodian plants of the Vernonia genus: Vernonia cinerea and Vernonia elliptica.An analytical HPLC method was developed and validated to quantify the major compound 8α-tigloyloxyhirsutinolide 13-O-acetate (H1) of Vernonia cinerea. We found that the compound H1 content ranges from 0.08% to 0.17% in samples collected from cambodian regions. We thus evaluated the anti-proliferative activity in vitro of extracts of V. cinerea and H1 compound against two human cancer cell lines HT29 and HepG2. For HepG2 cell line, the compound H1 has a similar IC50 to the positive control 5FU, 4.3 ± 0.2 versus 5.4 ± 0.9 µM. The interesting results on V. cinerea encouraged us to study other species of Vernonia genus growing in Cambodia, Vernonia elliptica. The phytochemical study conducted on this plant led to isolate a lignan [(+)-syringaresinol] and two sesquiterpene lactones [(+)-8,13-diacetyl-piptocarphol and Glaucolide B]. These compounds were described in this plant for the first time. The in vitro study of the anti-proliferative activity of the extracts of V. elliptica shows that the dichloromethane extract obtained from aqueous extract is very active.

The use and trade in medicinal plant products in South Africa is a cultivating sector of rural development. In order to conserve these plants and develop sustainable production, policies to promote domestication have been adopted. However, users view cultivated plants with scepticism. The key for policy makers is to understand how users respond to scarcity of species and whether domestication can be a solution. The main purpose of the study is to give an overview of variables that influence the rate of adoption of medicinal plant gardens. The last discussion will investigate whether domestication of species is an appropriate solution to conservation, or whether the intervention should be promoted as a local economic development option. The research took the form of a case study approach focusing on a specific area, with participatory and qualitative phases providing the basis for a semi-structured interview schedule, designed to assess the healer's acceptability of medicinal plant gardens, their comparative perception of cultivated and collected plants in their natural habitat/environment, and constraints in the establishment of medicinal plant gardens. Results indicated that:<li> The type of healer influences acceptability of medicinal plant gardens; •Healers use medicinal plant gardens to supply urban demand and provide income; • Healer and client perceptions of medicinal plant potency influence acceptability of cultivated plants; •Type of healer influences the environment in which species are collected. • Ancestral belief system forms an integral component of healers behavioural pattern.</li> The study concludes that:<li> Local scarcity of medicinal plants alone does not stimulate demand for nurseries; • Rituals associated with some treatments influence collection, storage and cultivation patterns; • Policy makers also need to promote domestication for commercial production and to investigate alternatives for propagating the population of some plant species in their natural habitat/environment to cater for specific needs of healers and • Interview responses, although useful, cannot always be taken at face value<//i>. / Dissertation (M Inst Agrar ( Agricultural Extension))--University of Pretoria, 2005. / Agricultural Economics, Extension and Rural Development / unrestricted

Advanced glycation end products the (AGE) comprise a series of compounds, quite different, whose activity is related to their ability to modify the chemical and functional properties of diverse biological structures. Several studies, including that used animals, indicate that AGE restriction in the diet can have many protective effects. Furthermore, the use of certain medicinal plants appears to offer additional benefits, especially to individuals suffering from disorders associated with excessive accumulation of AGE. It is believed that plants combiam antiglicantes and antioxidant properties, have positive effects on the inhibition of glycation. This fact has attracted the interest of the scientific community, as it can provide important therapeutic potential. Although any further component is particularly suitable as reducing AGEs in the context of the human body, some substances in vitro studies demonstrated the anti-AGE effects important and worth further investigation such as pyridoxamine, alilcisteína (garlic extract component), phenolics, vitamins C and E, thiamin, taurine and carnosine, recognized for its antioxidant properties. Therefore the aim of this proposal was to determine the antiglicante activity of extracts of plants native to the brasileleira flora of recognized antioxidant activity. All the studied extracts are part of the bank of plant extracts belonging to the Laboratory for Research on Natural Resources (LPqRN), the Institute of Chemistry and Biotechnology - UFAL. Therefore, this thesis was divided into three parts, which generated three articles. The first titled "Antioxidant activity of native plant extracts of flora by ABTS methods, DPPH, FRAP and ORAC", aimed to investigate the antioxidant capacity of different plant extracts, by different in vitro assays (ABTS, DPPH, FRAP , ORAC), and determine whether there is correlation between the tested methods. In this proposal, 49 plant samples were studied, the tests of antioxidant capacity, DPPH; ABTS; FRAP and ORAC. Further tests were conducted to see if there was a correlation between the methods. The results were screened for to follow up the research, specifically, the antiglicante activity, believed to be related to the antioxidant activity. So the second article entitled "antiglicante in vitro activity of native plant extracts of flora", aimed to determine the antiglicante activity of native plant extracts of brazilian flora, with recognized antioxidant activity and the possibility of correlation between tests, antiglicante and antioxidants. For this 18 extracts, which had been classified as important sources of antioxidant compounds were tested for activity antiglicante. From the most promising extract, Auxemma oncocalyx Taub. (Bark) for activity, an isolated quinone thereof was tested in Oncocalixona A. This may be an important antiglicante agent, which resulted in the third section, "Oncocalyxone The functions of an anti-glycation agent in vitro", that this was submitted to the journal FOOD CHEMISTRY, ISSN: 0308-8146, Impact factor 3.259, on 02/12/14, in the form of Short communication. The particular this thesis is of importance, since investigations of this nature favors the development of research investigating potential therapeutic products, and that in turn may be useful in treating diseases associated with excessive accumulation of AGE / Os produtos de glicação avançada (AGE) englobam uma série de compostos, bastante diferentes entre si, cuja atividade está relacionada à capacidade destes de modificar as propriedades químicas e funcionais das mais diversas estruturas biológicas. Diversos estudos, inclusive que utilizaram animais, apontam que a restrição de AGE na dieta pode exercer diversos efeitos protetores. Além disso, o uso de determinadas plantas medicinais parece oferecer benefícios adicionais, especialmente, para indivíduos que sofrem com transtornos associados ao acúmulo exagerado de AGE. Acredita-se que plantas que combinam propriedades antiglicantes e antioxidantes, exercem efeitos positivos sobre a inibição da glicação. Este fato tem atraído o interesse da comunidade científica, visto que pode oferecer importante potencial terapêutico. Embora nenhum componente seja ainda especificamente indicado como redutor de AGEs no contexto do corpo humano, em estudos in vitro algumas substâncias demonstraram efeitos anti-AGE importantes e merecem investigações adicionais, tais como piridoxamina, alilcisteína (componente do extrato de alho), compostos fenólicos, vitaminas C e E, tiamina, taurina e carnosina, reconhecidos por suas propriedades antioxidantes. Por esta razão o objetivo da presente proposta foi determinar a atividade antiglicante de extratos de plantas nativas da flora brasileira, que tenham reconhecida atividade antioxidante. Todos os extratos estudados fazem parte do banco de extratos vegetais pertencentes ao Laboratório de Pesquisas em Recursos Naturais (LPqRN), do Instituto de Química e Biotecnologia – UFAL. Para tanto, a presente tese foi dividida em três partes, que geraram três artigos. O primeiro deles entitulado “Atividade antioxidante de extratos de plantas nativas da flora brasileira pelos métodos ABTS, DPPH, FRAP e ORAC”, teve como objetivo investigar a capacidade antioxidante de diferentes extratos de plantas, por diferentes ensaios in vitro (ABTS, DPPH, FRAP, ORAC), e determinar se existe correlação entre os métodos testados. Nesta proposta, foram estudadas 49 amostras vegetais, pelos ensaios de capacidade antioxidante, DPPH; ABTS; FRAP e ORAC. Adicionalmente foram realizados testes para verificar se existia correlação entre os métodos. Os resultados foram uma triagem para dar sequência às pesquisas, especificamente, da atividade antiglicante, que se acredita haver relação com a atividade antioxidante. Assim o segundo artigo entitulado “Atividade antiglicante in vitro de extratos de plantas nativas da flora brasileira”, teve como objetivo determinar a atividade antiglicante de extratos de plantas nativas da flora barasileira, com reconhecida atividade antioxidante e a possibilidade de correlação entre os testes, antiglicante e antioxidantes. Para isto 18 extratos, que haviam sido classificados como importantes fontes de componentes antioxidantes, foram testados para atividade antiglicante. A partir do extrato mais promissor, Auxemma oncocalyx Taub. (entrecasca) para a atividade, foi testada uma quinona isolada do mesmo, a Oncocalixona A. Esta demonstrou ser um importante agente antiglicante, o que resultou no terceiro artigo, “Oncocalyxone A functions as an anti-glycation agent in vitro”, este que foi submetido à revista FOOD CHEMISTRY, ISSN: 0308-8146, Fator de impacto 3.259, no dia 02.12.14, na forma de Short communication. A presente tese se reveste de particular importância, uma vez que investigações desta natureza favorecem o desenvolvimento de pesquisas que investiguem potenciais produtos terapêuticos, e que por sua vez, possam ser úteis no tratamento de doenças associadas ao acúmulo exagerado de AGE

This work describes the isolation and structural elucidation of some chemical constituents as well as antioxidant, anticholinesterasic and skin anti-inflammatory activities of extracts and some of the isolated compounds from Coutarea hexandra (Jacq.) K. Schum. (Rubiaceae). In the antioxidant assays, extracts from roots and leaves, when compared to positive controls used, free scavenger radical and inhibited the formation of peroxide of the linoleic acid. In the anticholinesterase and anti-inflammatory assays, at 0.6 mg/ear, some extracts from roots were effective in inhibiting the effects of the anticholinesterase enzyme and the formation of ear edema. The phytochemical investigation of some active extracts from roots resulted in the isolation of three phytosteroids (sitosterol, stigmasterol and sitostenone), a diterpene (2β,3α-dihydroxy-11-oxo-ros-5-ene), two triterpenes (2β,3α-dihydroxy-11,16-dioxo- 5-ene-octanor-cucurbitacin and 23,24-diidrocucurbitacin F 25-acetate) and five 4-phenylcoumarin derivatives [5,7,8-trimethoxy-4-(3,4-dihydroxyphenyl)coumarin, 5-hydroxy-7-methoxy-4-(p-hydroxyphenyl)-coumarin, 5,7,8-trimethoxy-4-(p-methoxyphenyl)coumarin, 5,7-dimethoxy-4-(3-hydroxy-4-methoxyphenyl)coumarin,and 5,7-dimethoxy-4-(p-methoxyphenyl) coumarin]. These compounds had their structures elucidated based on their NMR spectral data and by comparison with literature data. Among isolated compounds, 2β,3α-dihydroxy-11,16-dioxo-5-ene-octanorcucurbitacin and 2β,3α-dihydroxy-11-oxo-ros-5-ene are being reported for the first time; 5-hydroxy-7-methoxy-4-(p-hydroxyphenyl)coumarin are being described for the first time as natural product and 5,7,8-trimethoxy-4-(3,4-dihydroxy-phenyl)coumarin occur for the first time in the Coutarea genus. In the anti-inflammatory assays, 5,7-dimethoxy-4-(pmethoxyphenyl) coumarin, 5-hydroxy-7-methoxy-4-(p-hydroxyphenyl) coumarin, 5,7-dimethoxy-4-(3-hydroxy-4-methoxy-phenyl)coumarin and 23,24-dihydro-cucurbitacin F 25-acetate, at 0.6 mg/ear, inhibited the edema by 35, 67, 19 and 30%, respectively. In the DPPH assays, 5-hydroxy-7-methoxy-4-(p-hydroxyphenyl)coumarin (43.70 0.21 μg/mL) and 5,7,8-trimethoxy- 4-(3,4-dihydroxyphenyl)coumarin (20.85 0.53 μg/mL) free scavenger radical with IC50 values comparable to ascorbic acid and gallic acid, respectively. In the anticholinesterase assays, 2β,3α-dihydroxy-11-oxo-ros-5-ene showed inhibition of this enzyme. / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O presente trabalho descreve o isolamento e a elucidação estrutural de alguns constituintes químicos, bem como das atividades antioxidante, anticolinesterásica e antiinflamatória cutânea de extratos das raízes, folhas e cascas do caule de Coutarea hexandra (Jacq.) K. Schum. (Rubiaceae), bem como de algumas das substâncias isoladas. Nos ensaios para avaliar atividade antioxidante, extratos das raízes e folhas forneceram os melhores resultados e alguns dos extratos das raízes foram eficazes em inibir o efeito da enzima acetilcolinesterase e a formação do edema de orelha induzido pelo óleo de cróton, na concentração de 0,6 mg/orelha. A investigação fitoquímica efetuada com extratos das raízes resultou na obtenção de três fitoesteróides (sitosterol, estigmasterol e sitostenona), dois triterpenos (2β,3α-dihidroxi-11,16-dioxo-5-eno-octanorcucurbitacina e 25-acetato da 23,24-diidrocucurbitacina F), um diterpeno (2β,3α-diidroxi-11-oxo-ros-5-eno) e cinco derivados 4-fenilcumarínicos [5-hidroxi-7-metoxi-4-(p-hidroxifenil)cumarina, 5,7,8-trimetoxi-4-(pmetoxifenil) cumarina, 5,7-dimetoxi-4-(3-hidroxi-4-metoxifenil)cumarina, 5,7-dimetoxi-4-(pmetoxifenil) cumarina e 5,7,8-trimetoxi-4-(3,4-dihidroxifenil) cumarina]. Estes compostos tiveram suas estruturas elucidadas com base na análise dos dados de RMN e pela comparação com dados da literatura. Dentre as substâncias isoladas, o triterpeno 2β,3α-diidroxi-11,16-dioxo-5-eno-octanorcucurbitacina e o diterpeno 2β,3α-diidroxi-11-oxo-ros-5-eno estão sendo relatados pela primeira vez; a 5-hidroxi-7-metoxi-4-(p-hidroxifenil)cumarina está sendo descrita pela primeira vez como produto natural e a 5,7,8-trimetoxi-4-(3,4-diidroxifenil) cumarina ocorre pela primeira vez no gênero Coutarea. Nos ensaios antiinflamatórios, as substâncias 5,7-dimetoxi-4-(p-metoxifenil)cumarina, 5-hidroxi-7-metoxi-4-(p-hidroxifenil)cumarina e 25-acetato de 23,24-diidrocucurbitacina F inibiram o edema por cerca de 35, 67 e 30%, respectivamente. Nos ensaios frente ao DPPH, tanto a 5-hidroxi-7-metoxi-4-(p-hidroxifenil)cumarina (43,70 0,21 μg/mL) quanto a 5,7,8-trimetoxi-4-(3,4-diidroxifenil)cumarina (20,85 0,53 μg/mL) seqüestraram radicais livres com valores de CI50 comparáveis aos do ácido ascórbico e do ácido gálico, respectivamente. Nos ensaios anticolinesterásicos, o diterpeno 2β,3α-diidroxi-11-oxo-ros-5-eno inibiu a ação da enzima acetilcolinesterase.