Thesis (M.Sc. (Pharmacology)) -- Mahidol University, 1978. / Supported by the Faculty of Dentistry, Mahidol University and the National Research Council.

Thesis (MSc)--Stellenbosch University, 2013. / ENGLISH ABSTRACT: In traditional medicine, there is a long-standing culture of combining herbal drugs to increase the therapeutic efficacy. The improved medical action is thought to be due to synergistic interactions between different plant bioactive components. The aim of this study was to test the pharmacological interactions in a medicinal plant combination which consisted of Agathosma crenulata, Dodonaea viscosa and Eucalyptus globulus. The rationale for the analysis of this particular mixture is that it had noteworthy antibacterial activity and exhibited the highest activity out of seven medicinal plant mixtures previously investigated. Using chromatographic analysis, the phytochemistry of the plants was also assessed. The chloroform: methanol (1:1; v/v) extracts or hydo-distilled essential oils (A. crenulata and E. globulus) were screened individually and in combinations (double and triple plant combination) for activity against five respiratory pathogens using a microdilution assay. The antimicrobial interactions in combinations were assessed with the fractional inhibitory concentration (FIC) and the isobolograms. The organic extracts generally showed the highest antimicrobial activity with E. globulus having the highest activity with MIC values below 1 mg ml-1 representing noteworthy activity. The overall activity of the aqueous extracts was poor. The essential oil activity of E. globulus was mostly noteworthy (0.5 to 2 mg ml-1) while A. crenulata essential oil displayed moderate activity (1 to 4 mg ml-1). The ΣFIC values for double combinations (1:1) of A. crenulata with D. viscosa, A. crenulata with E. globulus and D. viscosa with E. globulus were calculated from the minimum inhibitory concentration (MIC) data and the interactions were classified as synergistic, additive, indifferent and antagonistic. The highest synergistic interactions observed were for a 1:1 combination of A. crenulata with E. globulus against K. pneumoniae, S. aureus and B. subtilis with ΣFIC values of 0.07. There was only one incident of antagonism noted in the study for D. viscosa with E. globulus (1:1) against C. neoformans with ΣFIC value of 4.25. The double combinations against selective pathogens (K. pneumoniae, S. aureus and E. coli) were further analysed for interactions using isobolograms. Mostly, the antimicrobial interactions as presented by the isobolograms were congruent with FIC results which further validated the occurrence of relevant antimicrobial interactions in those combinations. The ΣFIC values for triple combinations (1:1:1) revealed mostly synergistic interactions. When the triple combinations were analysed further against certain pathogens based on the predictions of the Design of Experiments software program (MODDE 9.1®), the MIC values remained the same despite the different combinations that were tested Thin layer chromatography (TLC) was used for a quick chemical fingerprinting of the plant extracts. This was followed by a bio-autographic assay. The chemical profiles of the organic extracts and essential oils from two of the study aromatic plants (A. crenulata and E. globulus) were further analysed with liquid chromatography mass spectrometry (LC-MS) and gas chromatography mass spectrometry (GC-MS) respectively. For combined plant extracts, a multivariate data analysis using principal component analysis (PCA) and hierarchical clustering analysis (HCA) was used to determine the relationship of the chemical make-up of combinations with that of individual plant extracts. According to the TLC analysis, E. globulus extracts had more compounds than the other two plants in the study. For the bio-autographic assay, E. globulus and combinations that included this plant showed greater inhibition zones than A. crenulata and D. viscosa. For the LC-MS analysis, PCA and HCA showed a close relationship between A. crenulata with D. viscosa, D. viscosa with E. globulus and the triple combination. Twenty one components were identified in the essential oil of A. crenulata representing 88.83% of the total oil composition. The oil was dominated by oxygen-containing monoterpenes (46.25%). In the essential oil of E. globulus, twenty six compounds were identified making up to 95.62% of the oil composition. Oxygen-containing monoterpenes (32.98%) also dominated the E. globulus essential oil. There was no great variation in essential oil metabolites of the individual plants and their combination as shown by both PCA and HCA. The enhanced in vitro antimicrobial activity and pharmacological interactions (synergy and additivity) in some of the combinations (double and triple) that were tested in this study adds scientific support to the use of medicinal plant combinations in Western Cape traditional medicine. The metabolic profiles of plants in combination might be unique due to interaction of the different plant bioactive molecules and thus result into defined antimicrobial activity. / AFRIKAANSE OPSOMMING: In tradisionele geneeskunde is dit ’n lank bestaande kultuur om kruiemiddels te kombineer om die terapeutiese werking daarvan te verhoog. Dié verbeterde mediese werking word toegeskryf aan die oënskynlik sinergistiese interaksies tussen verskillende bioaktiewe plantkomponente. Die doel van hierdie studie was om die farmakologiese interaksies in medisinale plantkombinasies van Agathosma crenulata, Dodonaea viscosa en Eucalyptus globulus te bestudeer. Daar is op die ontleding van hierdie spesifieke mengsel besluit omdat dit oor beduidende antibakteriese waarde beskik en omdat dit uit sewe medisinale plantmengsels wat voorheen bestudeer is, as die doeltreffendste een aangewys is. Die fitochemie van die plante is ook met behulp van chromatografiese ontleding beoordeel. Deur middel van ’n mikroverdunningstoets is die chloroform:metanol- (1:1; v/v-)ekstrakte of hidrogedistilleerde vlugtige olies (A. crenulata en E. globulus) individueel sowel as in kombinasie (dubbele en drievoudige plantkombinasies) nagegaan vir hul werking met betrekking tot vyf respiratoriese patogene. Die gekombineerde antimikrobiese interaksies is met behulp van fraksioneel stremmende konsentrasie (FIC) en isobologramme ondersoek. Die organiese ekstrakte het oor die algemeen die meeste antimikrobiese aktiwiteit by E. globulus getoon, met MIC-waardes onder 1 mg ml-1 wat as noemenswaardige aktiwiteit beskou is. Die algehele aktiwiteit van die waterekstrakte was swak. Die vlugtige-olieaktiwiteit van E. globulus was merendeels noemenswaardig (0,5 tot 2 mg ml-1), terwyl die vlugtige olie van A. crenulata matige aktiwiteit getoon het (1 tot 4 mg ml-1). Die ΣFIC-waardes vir dubbelkombinasies (1:1) van A. crenulata en D. viscosa, A. crenulata en E. globulus, en D. viscosa en E. globulus is uit die minimum stremmende konsentrasie (MIC) bereken en die interaksies is as sinergisties, additief, neutraal en antagonisties geklassifiseer. Die sterkste sinergistiese interaksies is by ’n 1:1-kombinasie van A. crenulata en E. globulus met betrekking tot K. pneumoniae, S. aureus en B. subtilis opgemerk, met ΣFIC-waardes van 0,07. Die studie het slegs een geval van antagonisme opgelewer, naamlik by D. viscosa en E. globulus (1:1) met betrekking tot C. neoformans, wat ’n ΣFIC-waarde van 4,25 geregistreer het. Die werking van die dubbelkombinasies met betrekking tot gekose patogene (K. pneumoniae, S. aureus en E. coli) is voorts met behulp van isobologramme vir interaksies nagegaan. Die antimikrobiese interaksies wat uit die isobologramme geblyk het, was meestal in pas met FIC-resultate, wat die bestaan van tersaaklike antimikrobiese interaksies in daardie kombinasies verder bevestig het. Die ΣFIC-waardes vir die drievoudige kombinasies (1:1:1) het meestal sinergistiese interaksies aan die lig gebring. Toe die drievoudige kombinasies verder op grond van die voorspellings van die sagteware Design of Experiments (MODDE 9.1®) met betrekking tot sekere patogene ontleed is, het die MIC-waardes onveranderd gebly, ondanks verskillende toetskombinasies. Dunlaagchromatografie (TLC) is vir ’n vinnige chemiese ontleding van die plantekstrakte gebruik en is gevolg deur ’n bio-outografiese toets. Die chemiese profiele van die organiese ekstrakte en vlugtige olies van twee van die aromatiese plante in die studie (A. crenulata en E. globulus) is verder met vloeistofchromatografie-massaspektrometrie (LC-MS) en gaschromatografie-massaspektrometrie (GC-MS) onderskeidelik ontleed. Vir gekombineerde plantekstrakte is veelveranderlike-ontleding in die vorm van hoofkomponentontleding (PCA) en hiërargiese groepsontleding (HCA) gebruik om die verhouding van die chemiese samestelling van kombinasies in vergelyking met dié van individuele plantekstrakte te bepaal. Volgens die TLC-ontleding beskik E. globulus-ekstrakte oor meer verbindings as die ander twee plante in die studie. Vir die bio-outografiese toets het E. globulus en kombinasies daarmee groter stremmingsones as A. crenulata en D. viscosa getoon. In die LC-MS-ontleding het PCA en HCA op ’n hegte verhouding tussen A. crenulata en D. viscosa, D. viscosa en E. globulus, en die drievoudige kombinasie daarvan gedui. Een-en-twintig komponente is in die vlugtige olie van A. crenulata gevind, wat 88,83% van die algehele oliesamestelling uitmaak. Die olie is deur suurstofhoudende monoterpene (46,25%) oorheers. Die vlugtige olie van E. globulus het 26 verbindings opgelewer, wat 95,62% van die oliesamestelling uitmaak. Suurstofhoudende monoterpene (32,98%) het ook die vlugtige olie van E. globulus oorheers. Nóg PCA nóg HCA het op enige beduidende variasie in die metaboliete van die vlugtige olies van die individuele plante en hul kombinasies gedui. Die verhoogde in vitro- antimikrobiese aktiwiteit en farmakologiese interaksies (sinergie en additiwiteit) in van die kombinasies (dubbel én drievoudig) wat in hierdie studie getoets is, bied wetenskaplike stawing vir die gebruik van medisinale plantkombinasies in Wes-Kaapse tradisionele geneeskunde. Die metaboliese profiele van plantkombinasies kan verander weens die interaksie van die verskillende bioaktiewe plantmolekules, en kan baie bepaalde antimikrobiese aktiwiteit tot gevolg hê.

Made available in DSpace on 2014-06-11T19:30:54Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-04-22Bitstream added on 2014-06-13T21:01:35Z : No. of bitstreams: 1 arruda_apccbn_me_botib.pdf: 476199 bytes, checksum: 0108cd81c3b674db160daded40713597 (MD5) / Universidade Estadual Paulista (UNESP) / A planta Serjania erecta Radlk (SAPINDACEAE) é uma espécie endêmica da região de cerrado e apresenta como indicação popular a utilização de suas folhas para o tratamento de úlcera. A partir destas informações etnofarmacológicas os extratos metanólico e clorofórmico de suas folhas foram avaliados quanto a sua ação gastroprotetora e tóxica. Os dois extratos não apresentaram atividade tóxica no modelo de toxicidade aguda e em triagem hipocrática em modelos de animais in vivo e ambos apresentaram atividade gastroprotetora em modelo de indução de úlcera gástrica por etanol absoluto. O extrato clorofórmico (apolar) apresentou maior atividade gastroprotetora do que o extrato metanólico o que instigou a caracterização do mecanismo de ação deste extrato vegetal. A caracterização do mecanismo de ação foi realizada através de ensaios biológicos com o objetivo de se avaliar a ação antisecretória (ligadura de piloro) e o envolvimento de grupamentos sulfidrílicos, de óxido nítrico e das terminações nervosas sensíveis à capsaicina na gastroproteção. Foi constatado que este extrato não apresenta atividade anti-secretória e sua ação gastroprotetora ocorre por envolvimento dos grupamentos sulfidrílicos, via ativação de óxido nítrico e também das terminações nervosas sensíveis à capsaicina. A caracterização fitoquímica do extrato metanólico aponta para a presença de saponinas, taninos, flavonóides e terpenos. Já o extrato clorofórmico de S. erecta possui saponinas, poliisoprenóides, triterpenos que podem estar envolvidos na ação gastroprotetora do extrato. Estes resultados mostram o potencial efeito antiulcerôgenico do extrato apolar das folhas de S. erecta frente aos agentes ulcerogênicos mais comuns ao homem, porém, ainda são necessários outros estudos para definir a classe fitoquímica predominante que podem estar diretamente relacionado ao efeito gastroprotetor deste extrato. / Serjania erecta Radlk (SAPINDACEAE) plant is an endemic species originally found in the Brazilian savanna region with a popular use indicated to ulcer treatment. The known ethnopharmacological data about this plant has directed this work to an evaluation of the methanolic and chloroformic extracts made out of its leaves considering their gastroprotective and toxic actions. Both extracts have showed no toxic activity in the acute toxicity model and the hippocratic screening, both conducted for in vivo animal models. These extracts have also showed a gastroprotective activity in the absolute ethanol model for gastric ulcer induction. Since the chloroformic extract (apolar) has presented a higher gastroprotective action when compared to the methanolic extract, its mechanism of action characterization was deeply studied. This mechanism of action characterization was conducted with biological tests aiming to evaluate the antisecretory effect (pylorus ligature) and the involvement of sulfhydryl groups, nitric oxide and capsaicin sensory nerves. The chloroformic extract does not show antisecretory effect as well as gastroprotective action involving sulfhydryl groups and the activation of both the nitric oxide and the capsaicin sensory nerves. The phytochemistry characterization of the methanolic extract points to the presence of saponins, tannins, flavonoids and terpenes. The chloroformic extract, however, presents saponins, polyisoprenoids and triterpenes which may be involved in the mentioned gastroprotective action. These results show the high potential anti-ulcerogenic effect of the S. erecta apolar extract against the most common ulcerogenic agents for human beings. Additional studies are necessary in order to define the predominant phytochemistry class that may be directly related to the gastroprotective effect of this extract.

This dissertation is based upon research of an emergent ethnomedicine in a botanically rich area, the Western Cape of South Africa. It examines the interface between ethnobotany and medical anthropology by investigating the biological and social factors related to the knowledge, use and trade of medicinal plants by Cape bush doctors. Incredible syncretism was observed in the identity formation of this homogenous urban group of healers who combined elements of a globally recognized eco-religion and sociopolitical movement Rastafari with several South African cultures through knowledge of medicinal plants. By rejecting colonial principals, including capitalistic biomedical systems, bush doctors have crafted a niche acquiring knowledge and herbal remedies for the treatment of common ailments. Transmission amongst Rastas and trail-and-error experimentation with herbs emphasize that plant knowledge is situational and arises through relationships. From an estimated 200 bush doctors in the Cape, 62 almost exclusively middle-aged coloured males were interviewed. They declared their mission was 'to heal all people' through a reintroduction of KhoiSan healing traditions, an indigenous ancestry largely rejected by coloureds during apartheid. An ethnobotany of bush doctors revealed that of 192 species, 181 were medicinal and included various herbs important to most South African cultural groups. This largely herbaceous pharmacopeia is narrow compared to the region's high biodiversity and thirty-three species were identified as conservation priorities. The presence of bush doctors at transportation hubs as herbal hawkers creates a diversified economy through cultivation of relationships with primarily disadvantaged coloured and black consumers. The consumption, trade and sale of local plants by bush doctors represent efforts to embody the landscape; it reasserts coloured links to indigeneity, renews respect for their heritage and affords rights to resources. By evoking tradition within their tolerant philosophy, leaders of this emergent ethnomedicine develop a racially equitable and ecologically sustainable platform for health care evidenced by medicinal plant gardens in townships and transmission of diverse ethnomedical knowledge. Bush doctors are legitimized through the performance of transmission. This phytomedicinal knowledge reifies an ideology, repositioning coloureds in a moderate stance between colonial biomedicine and traditional African ethnomedicines, but also creates a unified South African medicine.

Orientador: Clélia Akiko Hiruma-Lima / Banca: Luis Cláudio Di Stati / Banca: Roberto L. Coelho / Resumo: A planta Serjania erecta Radlk (SAPINDACEAE) é uma espécie endêmica da região de cerrado e apresenta como indicação popular a utilização de suas folhas para o tratamento de úlcera. A partir destas informações etnofarmacológicas os extratos metanólico e clorofórmico de suas folhas foram avaliados quanto a sua ação gastroprotetora e tóxica. Os dois extratos não apresentaram atividade tóxica no modelo de toxicidade aguda e em triagem hipocrática em modelos de animais in vivo e ambos apresentaram atividade gastroprotetora em modelo de indução de úlcera gástrica por etanol absoluto. O extrato clorofórmico (apolar) apresentou maior atividade gastroprotetora do que o extrato metanólico o que instigou a caracterização do mecanismo de ação deste extrato vegetal. A caracterização do mecanismo de ação foi realizada através de ensaios biológicos com o objetivo de se avaliar a ação antisecretória (ligadura de piloro) e o envolvimento de grupamentos sulfidrílicos, de óxido nítrico e das terminações nervosas sensíveis à capsaicina na gastroproteção. Foi constatado que este extrato não apresenta atividade anti-secretória e sua ação gastroprotetora ocorre por envolvimento dos grupamentos sulfidrílicos, via ativação de óxido nítrico e também das terminações nervosas sensíveis à capsaicina. A caracterização fitoquímica do extrato metanólico aponta para a presença de saponinas, taninos, flavonóides e terpenos. Já o extrato clorofórmico de S. erecta possui saponinas, poliisoprenóides, triterpenos que podem estar envolvidos na ação gastroprotetora do extrato. Estes resultados mostram o potencial efeito antiulcerôgenico do extrato apolar das folhas de S. erecta frente aos agentes ulcerogênicos mais comuns ao homem, porém, ainda são necessários outros estudos para definir a classe fitoquímica predominante que podem estar diretamente relacionado ao efeito gastroprotetor deste extrato / Abstract: Serjania erecta Radlk (SAPINDACEAE) plant is an endemic species originally found in the Brazilian savanna region with a popular use indicated to ulcer treatment. The known ethnopharmacological data about this plant has directed this work to an evaluation of the methanolic and chloroformic extracts made out of its leaves considering their gastroprotective and toxic actions. Both extracts have showed no toxic activity in the acute toxicity model and the hippocratic screening, both conducted for in vivo animal models. These extracts have also showed a gastroprotective activity in the absolute ethanol model for gastric ulcer induction. Since the chloroformic extract (apolar) has presented a higher gastroprotective action when compared to the methanolic extract, its mechanism of action characterization was deeply studied. This mechanism of action characterization was conducted with biological tests aiming to evaluate the antisecretory effect (pylorus ligature) and the involvement of sulfhydryl groups, nitric oxide and capsaicin sensory nerves. The chloroformic extract does not show antisecretory effect as well as gastroprotective action involving sulfhydryl groups and the activation of both the nitric oxide and the capsaicin sensory nerves. The phytochemistry characterization of the methanolic extract points to the presence of saponins, tannins, flavonoids and terpenes. The chloroformic extract, however, presents saponins, polyisoprenoids and triterpenes which may be involved in the mentioned gastroprotective action. These results show the high potential anti-ulcerogenic effect of the S. erecta apolar extract against the most common ulcerogenic agents for human beings. Additional studies are necessary in order to define the predominant phytochemistry class that may be directly related to the gastroprotective effect of this extract / Mestre

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2018-11-05T13:39:32Z No. of bitstreams: 2 Dissertação - Luciana Oliveira Barateli - 2018.pdf: 3641421 bytes, checksum: a96d12f5fce8d5c02a9793dec50ebfbf (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2018-11-05T14:49:58Z (GMT) No. of bitstreams: 2 Dissertação - Luciana Oliveira Barateli - 2018.pdf: 3641421 bytes, checksum: a96d12f5fce8d5c02a9793dec50ebfbf (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2018-11-05T14:49:58Z (GMT). No. of bitstreams: 2 Dissertação - Luciana Oliveira Barateli - 2018.pdf: 3641421 bytes, checksum: a96d12f5fce8d5c02a9793dec50ebfbf (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2018-08-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Stryphnodendron adstringens presents a wide geographical distribution, being predominant in regions of Cerrado sensu stricto and is popularly known as "barbatimão". It is a species widely used in herbal medicine because of its anti-inflammatory, antibacterial and antiulcerogenic potential. The Cerrado’s removal of vegetation cover reduces significantly the number of several native species, among them S. adstringens. Thus, it is important that efficient strategies for the use and conservation of this species are drawn. In order to provide molecular tools for studies of genetic diversity and conservation of S. adstringens, the present work had the objective of developing microsatellite markers for the species. Genomic DNA was obtained from leaf tissue using the CTAB protocol. The identification of the microsatellite regions and the design of the primers were performed using the QDD program modules. From the identified microsatellite regions, 20 pairs of primers were designed, 14 of which flank microsatellite regions composed of dinucleotides, four by tetranucleotides and two by pentanucleotides. Initially, four individuals were used for the standardization tests of the PCR protocol and annealing temperatures. Subsequently, 48 individuals were selected, distributed in three populations, to evaluate polymorphism via 6% polyacrylamide gel. Of the 20 pairs of primers evaluated, 16 presented polymorphic amplification products and four monomorphic amplification products. Considering the 16 polymorphic markers, the number of alleles varied between two (SadH19) and 13 (SadH13), with a mean of seven alleles per locus. The observed heterozygosity (Ho) and expected (He) and PIC values were 0.506, 0.543, 0.635, respectively. The mean Hmax value founded (65,519) indicates values of genetic diversity that can be considered medians for this set of loci evaluated in three populations of S. adstringens. On the other hand, although genetic diversity is median, this set of 16 polymorphic markers exhibited a ombined probability of paternity exclusion high (0.9999983) and combined probability of genetic identity low (3,49x10-15). The analysis of variance of allelic frequencies presented significant values for two of the three estimated statistics with f not significant 0.050, significant θ equal to 0,329 and F in the overall value also significant 0.360. Thus, it can be concluded that the panel of polymorphic markers developed for S. adstringens is highly informative and indicated for population genetic studies for the species. Another important factor is that these markers can be tested in other evolutionarily close species for the availability of microsatellite markers, without the need to develop new primers. / Stryphnodendron adstringens apresenta ampla distribuição geográfica, sendo predominante em regiões de Cerrado stricto sensu e é conhecida popularmente como “barbatimão”. É uma espécie utilizada na medicina fitoterápica por seu potencial anti-inflamatório, antibacteriano e antiulcerogênico. A retirada de cobertura vegetal do Cerrado diminui significativamente o número de diversas espécies nativas, dentre elas S. adstringens. Dessa forma, é importante que sejam traçadas estratégias eficientes para o uso e a conservação dessa espécie. A fim de disponibilizar ferramentas moleculares para estudos de diversidade genética e conservação de S. adstringens, o objetivo desse estudo foi desenvolver marcadores microssatélites e estimar o potencial informativo desses marcadores para subsidiar estudos de genética populacional da espécie. A identificação das regiões microssatélites e o desenho dos primers foram realizados usando o programa QDD. Inicialmente, foram utilizados quatro indivíduos para os testes de padronização do protocolo de PCR e temperaturas de anelamento. Posteriormente, foram selecionados 48 indivíduos, provenientes de três populações, para avaliação de polimorfismo via gel de poliacrilamida a 6%. A partir das regiões microssatélites identificadas, foram desenhados 20 pares de primers, sendo que 14 flanqueiam regiões microssatélites compostas por dinucleotídeos, quatro por tetranucleotídeos e dois por pentanucleotídeos. Dos 20 pares de primers avaliados, 16 apresentaram produtos de amplificação polimórficos e quatro monomórficos. Considerando os 16 marcadores polimórficos, o número de alelos variou entre dois (SadH19) e 13 (SadH13), com uma média igual a sete alelos por loco. Os valores das médias da heterozigosidade observada (Ho) e esperada (He) e do PIC foram iguais a 0,506, 0,543, 0,635 respectivamente. A proporção da diversidade máxima encontrada foi de 65%, o que indica valores de diversidade genética que podem ser considerados medianos para esse conjunto de locos avaliados em três populações de S. adstringens. Por outro lado, embora a diversidade genética seja mediana, esse conjunto de 16 marcadores polimórficos exibiu uma probabilidade de exclusão de paternidade combinada alta (0,999983) e uma probabilidade de identidade combinada baixa (3,49x10-15). A análise de variância das frequências alélicas apresentou valores significativos para duas das três estatísticas estimadas com f não significativo 0,050, θ significativo igual a 0,329 e F no valor global também significativo 0,360. Assim, pode-se concluir que o painel de marcadores polimórficos desenvolvidos para S. adstringens é altamente informativo e indicado para estudos genéticos populacionais para a espécie. Outro fator importante é que esses marcadores podem ser testados em outras espécies próximas evolutivamente para possível disponibilização de marcadores microssatélites, sem a necessidade de desenvolvimento de novos primers.

Orientador: Alba Regina Monteiro Souza Brito / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Ciencias Medicas / Made available in DSpace on 2018-08-10T07:37:52Z (GMT). No. of bitstreams: 1 Miranda_MairaCola_D.pdf: 3726435 bytes, checksum: d25578678b3a9ed617824c5816dc825a (MD5) Previous issue date: 2006 / Resumo: O gênero Indigofera compreende 700 espécies herbáceas e arbustivas, figurando entre os seis maiores gêneros de Leguminosae. Em São Paulo foram descritas 9 espécies do gênero sendo que duas delas, Indigofera truxillensis Kunt e Indigofera suffruticosa Mill, são popularmente conhecidas como índigo, ocorrendo predominantemente em ambientes de Cerrado do Estado de São Paulo. A I. truxillensis é menos estudada do ponto de vista fitoquímico e farmacológico, sendo que a I. suffruticosa, possui atividade antiinflamatória, analgésica e digestiva indicadas por comunidades indígenas da América Central. Assim, o objetivo desse trabalho foi avaliar a atividade antiulcerogênica, das frações ricas em flavonóides, das espécies Indigofera truxillensis e Indigofera suffruticosa investigando os possíveis mecanismos de ação envolvidos com essa atividade. As frações aquosa (fAq) e acetato de etila (fAc) foram preparadas à partir do extrato metanólico (EMeOH) de ambas as espécies. O EMeOH das espécies foi utilizado para o ensaio de toxicidade aguda em camundongos. As frações fAq e fAc da I. truxillensis foram utilizadas nas doses de 50, 100 e 200 'mg.kg POT ¿1¿; já as frações fAq e fAc da I. suffruticosa, com um rendimento menor, foram utilizadas nas doses de 25, 50 e 100 'mg.kg POT ¿1¿, respectivamente. Ambas foram administradas pela via oral, em ratos. Para testar as frações das Indigoferas os modelos experimentais utilizados foram: úlcera gástrica induzida agudamente por etanol; estudo da atividade antisecretora através da ligadura do piloro; avaliação de mecanismos protetores da mucosa gástrica como a produção de muco, prostaglandina E2 (PGE2), envolvimento do óxido nítrico (ON) e compostos sulfidrilas (SH) na gastroproteção; avaliação da atividade antioxidante através do modelo de úlcera gástrica induzida por isquemia e reperfusão e atividade de algumas das principais enzimas envolvidas no processo como, superóxido dismutase (SOD), glutationa peroxidase (GPx) e glutationa redutase (GR); e avaliar a atividade na cicatrização de úlcera gástrica através do modelo de úlcera gástrica induzida por ácido acético (subcrônico) com posterior análise histológica para produção de muco e ensaios de imunohistoquímica para Heat Shock Protein (HSP 70) e Proliferação celular (PCNA). Os extratos das espécies estudadas demonstraram ausência de toxicidade (in vivo). A fAq e fAc da I. truxillensis e fAc da I. suffruticosa reduziram significativamente as lesões gástricas; a fAq da I. suffruticosa não apresentou atividade antiulcerogênica. Não ocorreu alteração no pH do suco gástrico, indicando que as frações das espécies estudadas não promovem atividade antisecretora. Nos mecanismos envolvendo proteção, a fAq da I. truxillensis protege a mucosa gástrica através do aumento de muco; envolvimento de ON e SH; aumento da atividade das enzimas SOD e GR; e expressão de HSP 70 e PCNA auxiliando no processo de cicatrização. Já a fAc da mesma espécie protege a mucosa gástrica através do aumento de muco e envolvimento de SH; aumento da atividade das enzimas SOD, GPx e GR; redução significativa na área da lesão na úlcera subcrônica. Nos mecanismos envolvendo proteção, a fAc da I. suffruticosa protege a mucosa gástrica através do aumento de muco; envolvimento de SH; aumento da atividade das enzimas GR; redução significativa na área da lesão na úlcera subcrônica e expressão de HSP 70 e PCNA auxiliando no processo de cicatrização. Uma triagem fitoquímica preliminar indicou a presença de flavonóides e alcalóides como um dos componentes majoritários na I. truxillensis e na I. suffruticosa. Através da análise dos resultados podemos concluir que a atividade gastroprotetora das espécies estudadas pode ser atribuída à presença de substâncias como os flavonóides, conhecidas por apresentarem atividade antiulcerogênica, antiinflamatória e antioxidante / Abstract: The genus Indigofera, which includes 700 herbaceous and bush species, is one of the six largest Leguminosae genres. In São Paulo State, Brazil, nine species of this genre were already described. Indigofera truxillensis Kunt and Indigofera suffruticosa Mill, commonly known as ¿índigo¿, are found predominately at São Paulo State¿s savannah. There are just a few studies on I. truxillensis pharmacology and phytochemistry, whereas I. suffruticosa presents anti-inflammatory, analgesic and digestive proprieties, cited by many Indian communities at Central America. The objective of this work is to evaluate the antiulcerogenic activity of I. suffruticosa and I. truxillensis fractions, which are rich in flavonoids, and also to investigate the possible action mechanisms involved in this activity. The aqueous (fAq) and ethyl acetate (fAc) fractions were obtained from methanol extract (EMeOH) from both species. The EMeOH was used in the acute toxicity study in mice. The I. truxillensis fAq and fAc were used at doses 50, 100 and 200 'mg.kg POT ¿1¿, and the I. suffruticosa fAq and fAc were used at doses 25, 50 and 100 'mg.kg POT ¿1¿. Both of them were administrated p.o., in male Wistar rats. In order to test the fractions, the experimental models used were the following: a) Gastric ulcer induced by ethanol; b) Antisecretory activity study through pylorus ligature; c) Evaluation of the protective mechanisms of gastric mucosal such as mucus production, prostaglandin E2 (PGE2), nitric oxide evolvement, and sulphydryl compounds (SH) in gastroprotection; d) Evaluation of antioxidant activity using gastric ulcer induced by ischemic and reperfusion, and some of the most important enzymes involved in the process, such as superoxide dismutase (SOD), gluthatione peroxidase (GPx) and gluthatione redutase (GR); e) Evaluate the gastric ulcer healing activity using the model of gastric ulcer induced by acid acetic, with subsequent histological analysis to mucus production and immunohistochemistry assay to Heat Shock Protein (HSP 70) and Cell Proliferation (PCNA). The crude extracts of both species demonstrated the absence of in vivo toxicity. The I. truxillensis fAq and fAc and the I. suffruticosa fAc reduced significantly the gastric lesions. The I. suffruticosa fAq did not present any antiulcerogenic activity. The gastric juice pH did not modify, which indicates no antisecretory activity for the fractions. I. truxillensis (fAq) protected the gastric mucosa through the increase of mucus; involvement of nitric oxide and SH; increase in the enzymes SOD and GR activity; expression of HSP 70 and PCNA in healing process. I. truxillensis (fAc) protected the gastric mucosa through the increase of mucus; involvement of SH; increase in the enzymes SOD, GPx and GR activity; significant reduction on subchronic ulcer lesion area. I. suffruticosa (fAc) protected the gastric mucosa through the increase of mucus; involvement of SH; increase in the enzyme GR activity; significant reduction on subchronic ulcer lesion area; expression of HSP 70 and PCNA in healing process. A preliminary phytochemistry selection and chromatographic analyses indicate the presence of flavonoids as majority compounds in I. truxillensis, and flavonoids and alkaloids in I. suffruticosa. From the analysis of the results it can be concluded that the gastroprotective activity of the studied species can be attributed to the presence of flavonoids, well known to present antiulcerogenic, anti-inflammatory and antioxidant activities / Doutorado / Doutor em Farmacologia

Submitted by (edna.saturno@ufrpe.br) on 2016-06-14T14:18:23Z No. of bitstreams: 1 Alejandro Lozano Balcazar.pdf: 872682 bytes, checksum: e3abd135cdb60662dc0bab0b46db32ec (MD5) / Made available in DSpace on 2016-06-14T14:18:23Z (GMT). No. of bitstreams: 1 Alejandro Lozano Balcazar.pdf: 872682 bytes, checksum: e3abd135cdb60662dc0bab0b46db32ec (MD5) Previous issue date: 2009-06-15 / This search analyzes a pharmacopoeia under the optics of the apparency hypothesis. This hypothesis divides plants into apparent and not apparent ones, being the first, in a simple way, perennial woody plants, and the second herbaceous with short life cycle. The apparency hypothesis, when facing the ethnobotany, proposes a positive relationship between the availability of species (usually measured by phytosociological parameters) and its use value for people (calculated through ethnobotanical information). In the community of Caçimbas, settlement which uses plant resources from the Cerrado of the Araripe National Forest (FLONA-Araripe) in the State of Ceará, semi-structured interviews were used to obtain the inventory of medicinal plants, their collection site, diseases for which they are indicated, and calculate its use value, as well as their commercial and practical importance. During the process of botanical collections was set the habit of the plants and they also were featured into woody and non woody ones. The herbaceous habit and the woody plants had the greatest richness of etnoespécies, the herbaceous and non woody ones had the highest practical value, and anthropogenic areas resulted to be the main source of medicinal plants; herbs and trees had equal versatility in the treatment of diseases and they do not differ in regards to the use value. The trees stood out as the most commercially important. In addition, two vegetation plots were held at FLONA to test the relationship between phytosociological parameters and the use and commercial value of the medicinal plants. The commercial and use value were not correlated with the abundance, frequency or ecological importance index of the plants. This study provides information for the development and management plan of the Araripe National Forest, indicating how is the dynamic of use of medicinal plants resources by a nearby community, and allowing the analysis of the role of the FLONA in the cultural, practical and commercial dynamics of use of medicinal plants. This survey provides data on the use of medicinal plants in Brazilian semiarid, arriving with information about the application of the apparency hypothesis in understanding the dynamics of use of these resources. / Esta pesquisa foi baseada na análise de uma farmacopéia sob a ótica da hipótese da aparência. Esta hipótese divide as plantas em aparentes e não aparentes, sendo, simplificadamente, as primeiras lenhosas perenes e as segundas herbáceas de ciclo de vida curto. A hipótese da aparência, quando virada para a etnobotánica, propõe uma relação positiva entre a disponibilidade das espécies (usualmente medida por parâmetros fitossociológicos) e o seu valor de uso para as pessoas (calculado por meio de informações etnobotânicas). Na comunidade de Caçimbas, povoado que utiliza recursos vegetais no cerrado da Floresta Nacional do Araripe (FLONA-Araripe) estado do Ceará, foram utilizadas entrevistas semiestruturadas para obter o inventário das plantas medicinais, seu local de coleta, doenças para as que são indicadas, e calcular seu valor de uso, valor comercial e importância prática. Durante coletas botânicas foi definido o hábito das plantas e também foram caracterizadas em lenhosas e não lenhosas. O hábito herbáceo e as lenhosas tiveram a maior riqueza de etnoespécies, as não lenhosas herbáceas são as de maior valor prático, e as zonas antropogênicas resultaram ser a principal fonte de plantas medicinais; ervas e árvores resultaram igualmente versáteis no tratamento de doenças e não se diferenciaram quanto ao valor de uso. As árvores se destacaram como as de maior importância comercial. Além disso, duas parcelas de vegetação foram realizadas na FLONA para testar a relação entre parâmetros fitossociológicos e o valor de uso e valor comercial das plantas medicinais. O valor de uso e valor comercial não se correlacionaram com a abundancia, freqüência e índice de valor de importância ecológico das plantas medicinais. O presente estudo fornece informação para o desenvolvimento do plano de manejo da Floresta Nacional do Araripe (FLONA-Araripe) indicando como é a dinâmica de uso de plantas medicinais por uma comunidade adjacente, e possibilitando a análise do papel da FLONA no valor cultural e importância prática e comercial das plantas medicinais. Esta pesquisa fornece dados sobre o uso de plantas medicinais no semiárido brasileiro, aportando com informações sobre a aplicação da hipótese da aparência na compreensão da dinâmica de uso desses recursos.

Thesis (M.Sc.(Biochemistry)) -- University of Limpopo, 2020 / Metastatic cancer remains incurable and accounts for 90% of cancer-related deaths (He et al., 2019). Therefore, there’s an urgent need to find anti-metastatic drugs with novel therapeutic targets (Zhang et al., 2018). Medicinal plants are promising sources of novel compounds with anti-metastatic activity (Tungsukruthai et al., 2018). This study investigated the anti-metastatic effects of Momordica balsamina L. crude acetone leaf extract in human HT-29 colon cancer cells. Powdered leaves of M. balsamina were macerated in acetone and reconstituted in dimethyl sulfoxide (>99.9%). The cytotoxic effect of the M. balsamina extract was investigated using the MTT assay. The acridine orange/ethidium bromide dual staining assay was used to show that the chosen concentrations of the M. balsamina extract do not induce apoptosis. The effect of the M. balsamina extract on reactive oxygen species formation and epithelial to mesenchymal transition-related morphological changes were assessed by the DCFH2-DA assay and light microscopy, respectively. The anti-invasive, anti-migratory and anti-adhesive effects of the M. balsamina extract were investigated using the cell invasion, wound-healing and cell adhesion assays, respectively. The adhesion of HT-29 cells to collagen I, II and IV, fibronectin, laminin, tenascin C and vitronectin was assessed using the ECM-cell adhesion array kit. Furthermore, western blotting was used to assess the effect of the M. balsamina extract on the expression of TNF-α, NF-κB, MMP-2, MMP-9 and TIMP-3. The findings revealed that the M. balsamina extract significantly inhibited the viability of HT-29 cells at concentrations above 50 µg/ml but had no effect on the viability of C3A liver cells at 40 and 80 µg/ml. Apoptotic features such as cell shrinkage, nuclei condensation, loss of membrane function and formation of apoptotic bodies were observed at 48 hours exposure to the M. balsamina extract. Reactive oxygen species formation, epithelial to mesenchymal transition, invasiveness, migration and adhesion were suppressed in HT-29 cells treated with the M. balsamina extract for 24 hours. The adhesion of HT-29 cells were varied amongst different extracellular matrix proteins. Furthermore, HT-29 cells treated with the M. balsamina extract showed a reduction in the expression of TNF-α, NF-κB, MMP-2 and MMP-9 proteins and an upregulation of TIMP-3 protein. In conclusion, the M. balsamina extract tested in this study impedes the metastatic cascade in HT-29 colon cancer cells by inhibiting the reactive oxygen species-mediated TNF-α/NF-κB/MMP-2/MMP-9 pathway. The findings suggest that M. balsamina L. may be a source of compounds with potential therapeutic use for the treatment of metastatic colon cancer. / National Research Foundation (NRF) South African Medical Research Council (SAMRC)

MSc (Chemistry) / Department of Chemistry / Bridelia micrantha (Hochst) Baill (B. micrantha) is a South African medicinal plant used by traditional healers in the treatment of different human ailments including diabetes, gastrointestinal ailments, joint aches, cough, conjunctivitis, skin problems and malaria. Previous studies have demonstrated the antidiabetic activities of B. micrantha crude extracts in in vivo studies. However, there are no studies on the compounds responsible for the antidiabetic activity of the plant. The purpose of this study was to isolate and characterize the antidiabetic constituents from B. micrantha. Materials and methods Crude methanolic extracts of root, stem and leaf were investigated using in vitro antidiabetic enzyme assays. Antioxidant activities were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing power assays. The isolation of the antidiabetic constituents was carried out using column chromatography on silica gel. Purification of the active fraction was carried out using preparative thin layer chromatography (pTLC). Structure elucidation of the compound was carried out using Nuclear Magnetic Resonance (NMR) spectroscopy and by comparison with literature. Results The results obtained in this study indicated that root, stem and leaf extracts exhibited high inhibition activity against α-glucosidase (98.52, 98.62 and 81.62% respectively). A moderate inhibition against α-amylase enzyme was observed for root (65.62%) and stem (61.86%) extracts. Leaf fraction LFR5 exhibited a high inhibition activity of 96.19% against α-glucosidase. Moreover, the isolated compound showed 96.74% inhibition against α-glucosidase. DPPH results revealed that antioxidant activity of crude extracts was not significantly different and they were concentration-dependent. Reducing power results revealed that stem (119.31 μg/mL) extract had higher activities compared to root (125.17 μg/mL) and leaf (291.88 μg/mL) extracts. Conclusion Quercetin-3-O-β-D-glucopyranosyl-(1→4)-α-L-rhamnoside was successfully isolated from B. micrantha leaves. Furthermore, quercetin-3-O-β-D-glucopyranosyl-(1→4)-α-L-rhamnoside demonstrated the ability to inhibit significantly the carbohydrate hydrolysing enzyme α-glucosidase and therefore validate the ethnomedicinal use of B. micrantha in the management of diabetes / Bridelia micrantha (Hochst) Baill (B. micrantha) is a South African medicinal plant used by traditional healers in the treatment of different human ailments including diabetes, gastrointestinal ailments, joint aches, cough, conjunctivitis, skin problems and malaria. Previous studies have demonstrated the antidiabetic activities of B. micrantha crude extracts in in vivo studies. However, there are no studies on the compounds responsible for the antidiabetic activity of the plant. The purpose of this study was to isolate and characterize the antidiabetic constituents from B. micrantha. Materials and methods Crude methanolic extracts of root, stem and leaf were investigated using in vitro antidiabetic enzyme assays. Antioxidant activities were evaluated using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and ferric reducing power assays. The isolation of the antidiabetic constituents was carried out using column chromatography on silica gel. Purification of the active fraction was carried out using preparative thin layer chromatography (pTLC). Structure elucidation of the compound was carried out using Nuclear Magnetic Resonance (NMR) spectroscopy and by comparison with literature. Results The results obtained in this study indicated that root, stem and leaf extracts exhibited high inhibition activity against α-glucosidase (98.52, 98.62 and 81.62% respectively). A moderate inhibition against α-amylase enzyme was observed for root (65.62%) and stem (61.86%) extracts. Leaf fraction LFR5 exhibited a high inhibition activity of 96.19% against α-glucosidase. Moreover, the isolated compound showed 96.74% inhibition against α-glucosidase. DPPH results revealed that antioxidant activity of crude extracts was not significantly different and they were concentration-dependent. Reducing power results revealed that stem (119.31 μg/mL) extract had higher activities compared to root (125.17 μg/mL) and leaf (291.88 μg/mL) extracts. Conclusion Quercetin-3-O-β-D-glucopyranosyl-(1→4)-α-L-rhamnoside was successfully isolated from B. micrantha leaves. Furthermore, quercetin-3-O-β-D-glucopyranosyl-(1→4)-α-L-rhamnoside demonstrated the ability to inhibit significantly the carbohydrate hydrolysing enzyme α-glucosidase and therefore validate the ethnomedicinal use of B. micrantha in the management of diabetes.