Lippia genus has a great pharmacological and therapeutic potential and encompasses approximately 200 species of herbs, bushes and small trees with a consistent chemical composition and pharmacological, therapeutics and culinary activities. This research aimed to evaluate the chemical and pharmacological aspects of Lippia gracilis Schauer (Verbenaceae), well-known as alecrim do campo , which grew at the germplasm bank of the Federal University of Sergipe, Brazil. This study was divided in two parts: in the first one we tried to evaluate the nonpolar compounds from the essential oil (EO) of L. gracilis plants under hydric stress and without hydric stress. The identification of the EO was made by using GC/MS and its anti-inflammatory and antinociceptive activities were studied in animal models. As results, thymol, p-cineme, methyl thymol, and carvacrol were found in the EO analysis, independent of the hydric stress L. gracilis leaves were submitted. Although the different hydric stresses the plants pass through, the chemical composition of the EO was too similar, so it was chosen to study the one obtained from the plants submitted to hydric stress in the animal models. In the animal models, we fed 1 hour before the beginning of the experiments with the EO (50, 100, and 200 mg/kg, p.o.), vehicle (tween 80 at 0.2% in saline, 10 mL/kg, p.o.) and acetylsalicylic acid (ASA, 300 mg/kg, p.o.) or dexamethasone (Dexa, 2 mg/kg, s.c.). To evaluate the anti-inflammatory activity, we used paw oedema model (Wistar rats, 120- 180 g) and leukocyte migration into the peritoneal cavity (Swiss mice, 20-30 g). To evaluate the antinociceptive activity, we used acetic acid-induced abdominal writhes model in Swiss mice. In all in vivo experiments we used groups of 6 animals. In the paw oedema assay, could be verified a reduction in inflammation (p<0.01) at the 200 mg/kg dose in Wistar rats in all measurement time (1, 2, 3 and 4 hours after carrageenaninduced paw oedema). The positive control (AAS 300 mg/kg) was also able to reduce the inflammation (p<0.05). In the leucocyte migration model, all EO doses used (50, 100, and 200 mg/kg) were able to reduces leucocytes migration, caused by carrageneen (p<0.01). Dexa was also able to reduce the migration (p<0.001). In the acetic acidinduced abdominal writhes model, all the doses reduced the nociception caused by acetic-acid, whereas the 200 mg/kg dose showed similar effects to ASS (p<0.001). In the second part of this study, the main polar compounds of L. gracilis were evaluated using antioxidant in vitro assays, namely ABTS (2,2 -azino-bis (3-ethylbenzothiazoline6- sulphonic acid), FRAP (Ferric Reducing Antioxidant Power) and DPPH (2,2-di(4- tertoctylphenyl)- 1-picrylhydrazyl), with trolox as reference antioxidant. The chemical composition of aqueous and ethanolic extracts was also evaluated by HPLC (High Performance Liquid Chromatography) and LCMS (Liquid Chromatography Mass spectrometry). Three major compounds with high antioxidant activity could be identified: luteolin-C-6-glucoside, luteolin-C-8-glucoside and carvacrol. The antioxidant activity of the ethanolic extract was higher than the water extract (about 40%), independent of the solvent used. These three compounds were also tested isolate, besides their comparison with well-known antioxidants as quercetin and luteolin. The antioxidant activity clearly depends on the assay used and the compound structure. Carvacrol showed higher activity in ABTS assay than C-6 and C-8-glucosides, but lower in DPPH and FRAP assays. In all assays, luteolin-C-6 showed higher antioxidant activity than luteolin-C-8-glucoside. L. gracilis is a good source of compounds with potential antioxidant, anti-inflammatory and antinociceptive activities. This is the first time that luteolin C-6 and C-8-glucosides were described in the L. gracilis. / O gênero Lippia apresenta grande potencial farmacológico e terapêutico e possui aproximadamente 200 espécies de ervas, arbustos e pequenas árvores, apresentando um perfil consistente de composição química e atividades farmacológicas, terapêuticas e culinárias. O objetivo deste trabalho foi estudar aspectos químicos e farmacológicos da Lippia gracilis Schauer (Verbenaceae), conhecida popularmente por alecrim do campo , proveniente do banco de germoplasma da Universidade Federal de Sergipe. Assim, este estudo foi dividido em duas partes: na primeira buscou-se estudar aspectos relacionados aos constituintes apolares da L. gracilis, presentes no óleo essencial (OE) obtido das folhas de plantas submetidas ao estresse hídrico (sazonal) e sem estresse. A composição dos constituintes do OE foi estudada com a utilização da cromatografia gasosa acoplada à espectrometria de massa (CG/EM) e suas atividades anti-inflamatória e antinociceptiva estudadas em modelos in vivo. Como resultados, verificamos que quatro compostos foram encontrados em maior concentração nos OE (independentemente das plantas terem sido submetidas ou não ao estresse hídrico): timol, p-cinemo, metil timol e carvacrol. Como a composição química dos OEs foi muito similar, escolheu-se trabalhar com o OE das plantas submetidas ao estresse hídrico nos ensaios in vivo. Para tanto, os animais (n=6/grupo) foram inicialmente prétratados com o OE (50, 100 e 200 mg/kg, v.o.), veículo (tween 80 a 0,2% em salina, 10 mL/kg, v.o.) e ácido acetilssalicílico (AAS, 300 mg/kg, v.o.) ou dexametasona (Dexa, 2 mg/kg, s.c.), dependendo do modelo experimental, 1 h antes do início dos experimentos. A atividade anti-inflamatória foi avaliada utilizando-se os modelos de edema de pata (ratos Wistar, 120-180 g) e peritonite (camundongos Swiss, 20-30 g) induzidos por carragenina. O tratamento dos animais com o OE na dose de 200 mg/kg reduziu de forma significativa (p<0,01) o edema induzido pela carragenina (1%, 100 μL/pata) em todos os tempos mensurados (1, 2, 3 e 4 h após a indução do edema). De forma similar, o AAS reduziu (p<0,05) a formação do edema em todos os tempos. No modelo de peritonite, todas as doses do OE (50, 100 e 200 mg/kg, p<0,01), bem como a dexametasona (p<0,001), foram capazes de reduzir a migração de leucócitos induzida por carragenina (1%, 250 μL, i.p.). Para avaliação da atividade antinociceptiva utilizouse o modelo de contorções abdominais induzidas por ácido acético (0,6%, 100 μL/10g, i.p.) em camundongos (Swiss, 20-30g). Observou-se que todas as doses do OE foram capazes de reduzir (p<0,05) a nocicepção induzida pelo ácido acético, sendo que com a dose de 200 mg/kg do OE observaram-se efeitos semelhantes ao AAS (p<0,001). Na segunda parte do presente estudo, os constituintes polares foram estudados in vitro, utilizando-se os ensaios de avaliação da atividade antioxidante do ABTS (2,2 -azino-bis (3-etil-benzolina-6-sulfonado), FRAP (poder antioxidante de redução do ferro) e DPPH (2,2-di(4-tertoctilfenil)-1-picrilhidrazil), com o composto de referência sendo o trolox, um análogo da vitamina E. A composição química dos extratos aquoso e alcoólico obtidos das folhas da L. gracilis, foi avaliada por cromatografia líquida de alto desempenho acoplada a espectrometria de massa (HPLC/EM). Através dos perfis químicos dos extratos obtidos por HPLC/EM, foi possível identificar três compostos majoritários com intensa atividade antioxidante: luteolina-C-6-glucosídeo, luteolina-C- 8-glucosídeo e carvacrol. Em ensaios in vitro, verificou-se uma maior atividade antioxidante (40%) do extrato alcoólico quando comparado ao aquoso, independente do solvente utilizado para a dissolução. Os três compostos majoritários foram testados isoladamente, além de terem sido comparados com compostos antioxidantes de ação conhecida, como quercetina e luteolina. As atividades detectadas dependeram claramente do ensaio utilizado e da estrutura do composto. O carvacrol apresentou maior atividade no ABTS, quando comparado com as luteolinas, porém atividade inferior no DPPH e no FRAP. Em todos os ensaios, a luteolina-C-6-glucosideo apresentou valores maiores que a luteolina-C-8-glucosideo. A L. gracilis representa uma fonte potencial de antioxidantes, além de possuir atividades anti-inflamatória e antinociceptiva. Este é o primeiro relato de detecção das luteolinas C-6 e C-8 glucosídeo na espécie Lippia gracilis.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / O efeito antibacteriano de Ãleos essenciais extraÃdos de folhas das espÃcies de Lippia aff. gracilis e Lippia gracilis frente a cepa de Staphylococcus aureus isolada de Ãlcera de paciente com pà diabÃtico foi avaliado mediante experimentaÃÃo in vitro e in vivo, utilizando-se o modelo experimental de ratos diabÃticos aloxano induzidos. Cento e doze ratos machos Wistar diabÃticos com peso mÃdio de 180g foram distribuÃdos ao acaso em dois experimentos. Cada experimento foi dividido em dois procedimentos que avaliaram a atividade antibacteriana de soluÃÃes dos Ãleos essenciais a 5%, em dois diferentes procedimentos: um no ato da administraÃÃo do inÃculo bacteriano e outro apÃs vinte e quatro horas de administraÃÃo. A administraÃÃo tanto do inÃculo 108ufc/ml quanto da suspensÃo dos Ãleos foi por via subcutÃnea no membro pÃlvico dos ratos diabÃticos. CinqÃenta e seis ratos Wistar foram utilizados para cada experimento, no qual os mesmos foram distribuÃdos ao acaso em 8 (oito) diferentes grupos: 4 grupos por experimento, apresentando 7 ratos por grupo (G1-Branco; G2-Controle negativo; G3-Controle positivo; G4-Teste). Foi verificado que no procedimento 1(S.aureus sem Lippia aff gracilis 108  698 versus S.aureus com Lippia aff gracilis 293,1  79,07; S.aureus sem Lippia gracilis 108  873 versus S.aureus com Lippia gracilis 302  57,2) e no procedimento 2 (S.aureus sem Lippia aff gracilis 108  313 versus S.aureus com Lippia aff gracilis 13,28  4,03; S.aureus sem Lippia gracilis 108  818 versus S.aureus com Lippia gracilis 13,14  4,27); houve reduÃÃo na contagem bacteriana tanto para Lippia aff. gracilis quanto para Lippia gracilis. Quando comparados os grupos G4 com G3, observou-se que esta suspensÃo a 5% nÃo apresentou efeito prÃ-inflamatÃrio. Para a validaÃÃo destes resultados, foram utilizados os testes Mann-Whitney e Bartletts &Newman-Keuls (MÃdia  - E.P.M) com nÃvel de significÃncia de (p<0,05). Ainda neste estudo foi avaliado a aÃÃo antioxidante e imunomoduladora da L-glutamina em modelos de ratos Wistar diabÃticos quando administrada por gavagem a uma concentraÃÃo de 0,7g/kg em um perÃodo de 30 dias. Quarenta ratos Wistar machos foram distribuÃdos ao acaso em 5 grupos (GI-nÃo diabÃtico; GII-diabÃtico; GIII-diabÃtico com salina; GIV-diabÃtico com L-glutamina; GV-diabÃtico com ProteÃna do Soro do Leite). Passado este perÃodo, foram determinadas as concentraÃÃes de substÃncias reativas ao Ãcido tiobarbitÃrico (TBARS) e glutationa reduzida (GSH) no soro e nos tecidos hepÃticos, pancreÃticos, mÃsculo esquelÃtico, rins e tecido adiposo. Para as comparaÃÃes entre o grupo tratado com L-glutamina com os demais, utilizou-se a anÃlise de variÃncia (ANOVA-teste Tukey). As comparaÃÃes entre grupos foram feitas utilizando-se o teste t de Student. Valores de p<0,05 foram considerados significantes. A suplementaÃÃo com L-glutamina induziu ao aumento nas concentraÃÃes de GSH (MÃdia  E.P.M) e reduÃÃo significante nas concentraÃÃes de TBARS (MÃdia  E.P.M), quando comparadas com o grupo controle, nos espÃcimes analisados. A aÃÃo imunomoduladora foi avaliada atravÃs da quantificaÃÃo de linfÃcitos CD4+ e CD8+ em sangue total. Vinte e quatro ratos Wistar machos diabÃticos foram distribuÃdos igualmente em 3 grupos (G1-diabÃtico com salina; G2-diabÃtico com L-glutamina; G3- diabÃtico com ProteÃna do Soro do Leite). Observou-se um aumento significante nas populaÃÃes de linfÃcitos CD4+ (MÃdia  E.P.M) com reduÃÃo nas populaÃÃes de linfÃcitos CD8+ (MÃdia  -E.P.M) do grupo G2 quando comparado com o grupo controle, ressaltando a importÃncia da L-glutamina como imunonutriente / The antibacterial effect of essential oils extracted from leaves of the species of Lippia aff. gracilis and Lippia gracilis against strains of Staphylococcus aureus isolated from patients with diabetic foot ulcers were evaluated in vitro and in vivo experimetation utilizing an experimental model of alloxan-induced diabetic rats. One hundred and twelve male diabetic Wistar rats with mean weight of 180g were distributed by chance into two experiments. Each experiment was divided in two procedures with evaluation of antibacterial activity of essential oils at 5% solutions; one procedure in the act of administration of bacterial inoculums and the other one 24 hours later. 108 CFU (Colony Forming Unit) /mL, as well as oils suspension inoculated in the subcutaneous tissue of the pelvic member of diabetic rats. Lower than 5% concentration of administered solution presented antibacterial effect in the in vitro experiment. Fifty-six Wistar rats were utilized in each experiment, randomly distributed in 08 different groups: 04 groups per experiment, each group with 07 rats (G1-White; G2-Negative Control; G3- Positive control; G4-Test). There was decrease in CFU/mL in procedure 1 (S.aureus without Lippia aff gracilis 108  698 versus S.aureus with Lippia aff gracilis 293,1  9,07; S.aureus without Lippia gracilis 108  873 versus S.aureus with Lippia gracilis 302Â57,2), which evaluated antibacterial effect of oils concomitantly with the administration of inocula as well as in procedure 2 (S.aureus without Lippia aff gracilis 108  313 versus S.aureus with Lippia aff gracilis 13,28  4,03; S.aureus without Lippia gracilis 108  818 versus S.aureus with Lippia gracilis 13,14  4,27) , which evaluated antibacterial effect of oils 24 hours after the administration of the inoculum . When comparing group G4 with G3, it was observed that 5% solution presented no pro-inflammatory effect, for analysis of these results, the tests of Mann-Whitney and Bartletts & Newman-Keuls (X  S.E.M) with level of significance (p<0,05) were used. Part of this study evaluated the antioxidant and immunomodulating effect of L-glutamine in models of Wistar diabetic rats when administered by gavages at a 0,7g/kg during 30 days. Fourty Wistar male rats were randomly distributed in 5 groups (GI- non diabetic; GII-diabetic; GIII-diabetic with saline; GIV- diabetic with L-glutamine; GV- diabetic with Whey Protein). After 30 days concentrations of TBARS and GSH in serum and in hepatic, pancreatic, skeletal muscles, kidneys and fat tissues were determined. For comparisons between the group treated with L-glutamine with the others, the ANOVA â Tukey test was utilized and the comparisons between groups were done using Studentâs t test. Values of p<0.05 were considered significant. The supplementation with L-glutamine induced an increase in the concentrations of GSH (Mean&#61617;S.E.M) and significant reduction in the TBARS (mean  S.E.M) concentrations, when compared to the control group, in the analyzed specimens. The immunomodulating effect was evaluated by quantification of CD4+ and CD8+ lymphocytes in total blood. Twenty-four Wistar male diabetic rats were distributed equally in 3 groups (G1-diabetic with saline; G2-diabetic with L-glutamine; G3- diabetic with Whey Protein). It was seen a significant increase of CD4+ (mean  S.E.M) lymphocytes with reduction of CD8+ (mean  S.E.M) lymphocytes in group G2 when compared to the control group, showing the importance of L-glutamine as an immunonutrient

An experiment was conducted to determine the effects of shade netting on growth, oil yield and compound composition of fever tea (Lippia javaica). The treatments were nine shade nets of different light intensities (100% bird net, 40% green, 10, 12 and 18% white and 30, 40, 55 and 70% black). The layout used was a completely randomized design. The parameters measured were shoot width, fresh shoot mass, the number of plants flowered, plant height and photosynthetically active radiation. Shade netting did not have any effect on the shoot width of L. javanica. Eighteen percent white net was best suited to produce fresh shoot mass, whereas 55% black net, 40% black net and 10% white net produced lower fresh shoot mass. More plants that flowered were obtained under 70% black net than those grown under 30% black net. The number of plants that flowered increased from 22 to 35 with an increase in shading density. The height of plants was not affected by shade netting at 49 DAT. However, at 59 and 67 DAT, the height of the plants was greater under low light intensities (40, 55, and 70% black nets) than that of high light intensity (30% black). Twelve percent white net had the greatest plant height at 102 DAT. Shade netting significantly affected the adsorption and utilization of photosynthetically active radiation (PAR) above, across and below the plants at 102 DAT. Hundred percent bird net had the greatest amount of light intercepted above, across and below the plants. The amount of light intercepted above, below and across the plants was lower for lower shading intensity (30% black) than that of higher shading intensities (40%, 55% and 70% black). To determine shade-netting effect on oil yield and compound composition, fresh leaves (± 200g) of L. javanica were sampled from each shade net for essential oil extraction by hydrodistillation. The oil yield ranged from 0.29% to 0.41%. The oil had a clear to yellowish brown colour. No significant differences were detected for oil yield amongst the shade nets. The chemical compound analysis was carried out by GC/MS. The chemical compounds identified in the essential oil were <font face="symbol">a</font>-pinene, sebinen, myrcene, 1.8 myrcene, ipsenone, ipsedienone, <font face="symbol">b</font>-caryophyllen and germacrene-D. L. javanica plants grown under 10% white net, resulted in significantly higher myrcene content (15.7%) as compared to plants grown in other shade nets. Shade netting did not have any effect on the composition of á-pinene, sebinene, 1.8 myrcene, ipsenone, ipsedienone, <font face="symbol">b</font>-caryophyllen and germacrene-D. For commercial usage of myrcene, white net (10%) would be best suited for L. javanica production. To obtain information about indigenous knowledge of L. javanica, a survey was carried out among street traders, traditional healers, hawkers and farmers in Gauteng. The age group was between 20 and 73. The majority of respondents, who were self employed, were traditional healers followed by farmers. Most (98%) of the respondents indicated that they collected L. javanica from the veld by cutting the plant, whereas other respondents uprooted the whole plant when harvesting. Amongst the different uses of L. javanica mentioned, treatment of colds and coughs through steaming was stated by most (47%) of the respondents. Most (88%) of the respondents felt that it was important to conserve the plant for future generations and also to prevent the plant from becoming extinct. Traditional healers knew the uses and other important aspects of L. javanica than the farmers. / Dissertation (M Inst Agrar (Agronomy))--University of Pretoria, 2007. / Plant Production and Soil Science / unrestricted

Fever tea is one of the important medicinal plants belonging to the family Verbenaceae. The leaves of the plant are used as a remedy to treat malaria, stomach pains, colds and fever. There are different clones of Lippia species available with different medicinal value. Therefore, the purpose of this study was to determine protocols for propagation of fever tea so as to multiply an ideal clone. Vegetative propagation of this plant species by stem cuttings, requirements for seed germination and response of fertigation frequencies and growing medium to growth, yield and quality has not been studied. In this study, factors influencing effective propagation of fever tea were studied. Those factors were: cutting position (apical vs. basal), media (pine bark vs. sand), hormone (seradix No. 2 vs. no hormone) light and temperature requirement for fever tea seed germination and effect of fertigation frequencies (0.4 L/day, 1L/day, 2L/day, 2L/2nd day and 2L/week) and growing media (pine bark vs. sand) on growth, oil yield and oil quality. In vegetative propagation by stem cuttings, measurements made were number of roots per rooted cutting, fresh mass, stem circumference and number of leaves and the rate of rooting during four consecutive sampling dates (5, 10, 15 and 20 days after planting). Apical cuttings rooted earlier by 5 days than basal cuttings at 10 days but at 15-20 days after establishment, both cuttings had good rooting. Fresh mass was not affected by cutting position and rooting media, but cuttings performed slightly better when propagated in pine bark medium than sand medium. Basal cuttings resulted in thicker stems and more leaves as compared to apical cuttings. Seradix No.2 (0.3% IBA) hormone increased the fresh mass, stem circumferences, root number and leaf numbers on both apical and basal cuttings. For the establishment of fever tea stem cuttings, both apical and basal cuttings can be used but pine bark is the ideal medium. The cuttings can be ready for transplanting in 15-20 days after establishment and Seradix No. 2 (0.3% IBA) promotes rooting of fever tea cuttings. The ideal combination of light and temperature for seed germination of fever tea was investigated. Germination was tested at constant temperature regimes (15, 20, 25 and 30¨¬C with continuous light or dark period and alternating temperatures of 20:30 and 16L: 8D (light: dark) combinations respectively. Seeds started to germinate after 8 days from incubation and the last germination was observed at 30 days from incubation. Germination percentages increased at 20-30 constant temperatures and 20/30 alternating temperatures but the difference amongst them was not significant. Higher germination of 86% were achieved when seeds were exposed to continuous light than alternating light and dark. Seeds failed to germinate in continuous darkness. Fever tea seeds were positively photoblastic. The effect of fertigation frequency and growing medium on the growth, yield and quality of fever tea were investigated in a tunnel. Treatments used were five fertigation frequencies (0.5L/day, 1L/day, 2L/day, 2L/2nd day, and 2L/week) and two growing media (pine bark and sand). Measurements made were plant height, stem circumference and number of branches at 8, 16 and 32 weeks after planting. At 8 weeks after planting all fertigation frequencies improved fever tea growth except fertigation frequency of 2L/week. All the fertigation frequencies were ideal to sustain the growth and development of fever tea plants except 2L/week. At 16 weeks after planting there were interactive effects between fertigation frequencies and the growing medium for the plant height of fever tea. Plants fertigated with 2L/day grown in sand media grew taller than all the other fertigation frequencies. At 32 weeks after planting there was a significant effect on the plant height from the main effects of fertigation frequency and growing medium. Plants fertigated with 2L/day were significantly the tallest followed by plants fertigated with 0.5L/day, 1L/day, 2L/2nd day and 2L/week. When plants were younger better plant growth was obtained in pine bark media. Stem circumference and number of branches of fever tea were significantly affected by fertigation frequency and growth medium. At 16 and 32 weeks after planting, plants grown in sand media had thicker stems and more branches as compared to plants grown in pine bark media. The essential oil of fever tea was extracted using hydro-distillation. Fertigation frequency did not affect oil yield. Plants grown in pine bark media yielded more oil than plants grown in sand medium. Microscopical studies using scanning electron microscope were investigated to determine the development of oil glands and trichomes on the abaxial (upper) and adaxial (lower) surfaces of the leaves as affected by fertigation frequency and the growing medium. There were no significant effects on the number of oil glands and trichomes developed on both surfaces of the leaves. Pine bark medium resulted in larger oil glands than sand medium regardless of the treatments, and pine bark also yielded more oil percentages than sand medium. Based on this investigations for commercial production of fever tea essential oil pine bark media is recommended. Chemical compounds of fever tea oil were also not affected by fertigation frequency or growth medium. In this study the chemical compounds detected from essential oils of fever tea were monoterpenes (i.e. ¥á-pinene, sebinen, 1.8 cinede, myrcene, ipsenone and ipsedienone) and the sesquiterpenes (i.e. ¥â-caryophyllene and germacrene-D). Compounds that gave the smallest chemical percentages and the shortest time to be detected were ¥á-pinene, sebinen, 1.8 cinede, myrcene, ipsenone and ipsedienone. Compounds found with the highest chemical percentages with highest peaks were ¥â-caryophyllene and germacrene-D. / Dissertation (MSc (Horticulture))--University of Pretoria, 2007. / Plant Production and Soil Science / unrestricted

Free-living amoebae (AVL), belonging to the genus Acanthamoeba are widely distributed in nature, being found in various types of soil and water, in the air, in air conditioning and sewage, and other environments. These amoebae are able to cause severe disease in human beings, highlighting the granulomatous amoebic encephalitis and keratitis. The amoebic keratitis is an infectious disease that affects the cornea and it can lead to blindness, with symptoms such as blurred vision, intense pain, photophobia and increased eye pressure. Users of contact lenses are more amenable to this type of infection because they may present microtraumatisms in the cornea, which makes the environment favorable action of these amoebae. There is no effective treatment for this disease and most used drugs suffers resistance from the parasite and others such as corticosteroids, they may give an initial improvement, but then the worsening of the disease. The solutions for cleaning contact lenses are also not efficient in eliminating this parasite, since the ISO 14729 and FDA (Food and Drug Administration, USA), which regulate the microbiological requirements and test methods for the production of hygiene products for contact lenses do not require that such solutions are tested against Acanthamoeba, which makes users more amenable to this type of lenses infection. In view of this, the objective of this study was to develop a microemulsion with the essential oil of L. gracilis and check its amoebicide potential front of trophozoites of Acanthamoeba castellanii. The formulations were prepared by using the phase diagram, using Tween 80 and propylene glycol as surfactant and co-surfactant of the system. The physico-chemical characterization of the formulations was performed by analysis of polarized light microscopy, rheology, light low angle scattering (SAXS), electrical conductivity, particle size and polydispersity. Amoebicide front activity assays for trophozoite A. castellanii formulation in question and all its separate excipients were performed. The obtained samples had transparent and thermodynamically stable to droplet size and characteristic polydispersity index microemulsion. The formulations presented are isotropic in polarized light microscopy and, through SAXS curves, it was observed that the formulations showed characteristics bicontinuous microemulsion. These formulations showed low electrical conductivity which can be associated with high amount of nonionic surfactant. The rheological properties of the system have shown that they possess newtonian behavior characteristic of microemulsion and that the viscosity increases when the amount of oil decreases and hence the droplet size. In bioassays, the surfactants showed no significant activity against the trophozoites and the essential oil of L. gracilis showed an IC50 of 9,52μg / ml. When the test was conducted with the microemulsion containing the same oil, this IC 50 fell to 2.55 μg / ml and using a formulation with smaller droplet tamnho this value fell to 0.65μg/ml, thus proving the efficacy of nanossistema. / Amebas de vida livre (AVL), pertencentes ao gênero Acanthamoeba, estão amplamente distribuídas na natureza, sendo encontradas em diversos tipos de solo e água, no ar, em ar-condicionado e esgotos, entre outros ambientes. Essas amebas são capazes de ocasionar graves doenças nos seres humanos, destacando a encefalite granulomatosa e a ceratite amebiana. A ceratite amebiana é uma doença infecciosa que acomete a córnea e que pode levar a cegueira, apresentando sintomas como visão borrada, dor intensa, fotofobia e aumento da pressão ocular. Os usuários de lentes de contato são os mais propícios a este tipo de infecção, pois podem apresentar microtraumatismos na córnea, o que torna o ambiente favorável a ação dessas amebas. Ainda não existe um tratamento eficaz para esta doença e a maioria dos medicamentos utilizados sofre resistência por parte do parasito e outros, como os corticosteróides, podem apresentar uma melhora inicial, mas em seguida o agravamento da doença. As soluções para limpeza das lentes de contato também não são tão eficientes em eliminar este parasita, uma vez que a ISO 14729 e a FDA (Food and Drug Administration, Estados Unidos), que regulamentam os requisitos microbiológicos e os métodos de ensaio para a produção de produtos de higiene para lentes de contato, não obrigam que essas soluções sejam testadas contra Acanthamoeba, o que torna os usuários de lentes mais propícios a este tipo de infecção. Diante disto, o objetivo deste trabalho foi desenvolver e caracterizar sistemas microemulsonados, contendo o óleo essencial de Lippia gracilis como fase oleosa e analisar sua potencial ação amebicida frente a trofozoítos de Acanthamoeba castellanii. As formulações foram preparadas através do uso do diagrama de fase, utilizando o Tween 80 e o propilenoglicol como tensoativo e cotensoativo, respectivamente, do sistema. A caracterização físico-química das formulações foi realizada por meio de análises de microscopia de luz polarizada, reologia, espalhamento de luz a baixo ângulo (SAXS), condutividade elétrica, tamanho de partícula e índice de polidispersão. Foram realizados ensaios de atividade amebicida frente à trofozoítos de A. castellanii da formulação em questão e de todos os seus excipientes separados. As amostras obtidas apresentaram-se transparentes e termodinamicamente estáveis com tamanho de gotícula e índice de polidispersão característicos de microemulsão. As formulações apresentaram-se isotrópicas na microscopia de luz polarizada e, através das curvas de SAXS, foi possível observar que as formulações apresentaram características de microemulsão bicontínua. Estas formulações mostraram baixa condutividade elétrica a qual pode estar associada a alta quantidade de tensoativo não iônico. As propriedades reológicas do sistema demonstraram que estes possuem comportamento newtoniano característico de microemulsão e que a viscosidade aumenta quando diminui a quantidade de óleo e, consequentemente, o tamanho da gotícula. Nos ensaios biológicos, os tensoativos não apresentaram atividade significativa frente aos trofozoítos e o óleo essencial de L. gracilis apresentou uma IC50 de 9,52μg/ml. Quando o teste foi realizado com a microemulsão contendo o mesmo óleo, essa IC50 caiu para 2,55 μg/ml e, ao utilizar uma formulação com tamanho de gotícula menor, esse valor caiu para 0,65 μg/ml, comprovando, assim, a eficácia do nanossistema. / São Cristóvão, SE

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-01-20T16:56:15Z No. of bitstreams: 1 flaviapereiracampos.pdf: 1279544 bytes, checksum: 9b6d679cec6fc27973877316210a3bf1 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-01-25T18:40:56Z (GMT) No. of bitstreams: 1 flaviapereiracampos.pdf: 1279544 bytes, checksum: 9b6d679cec6fc27973877316210a3bf1 (MD5) / Made available in DSpace on 2016-01-25T18:40:56Z (GMT). No. of bitstreams: 1 flaviapereiracampos.pdf: 1279544 bytes, checksum: 9b6d679cec6fc27973877316210a3bf1 (MD5) Previous issue date: 2014-04-28 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico / FAPEMIG - Fundação de Amparo à Pesquisa do Estado de Minas Gerais / As plantas do gênero Lippia pertencem à família Verbenaceae e caracterizam-se por possuir diversas espécies com propriedades medicinais. Dentre os aspectos químicos e farmacológicos investigados em Lippia, destacam-se aqueles relacionados aos seus óleos essenciais. Os óleos essenciais pertencem ao metabolismo secundário, são misturas complexas de substâncias voláteis e lipofílicas. O objetivo deste estudo foi investigar efeitos citogenotóxicos de óleos essenciais extraídos de Lippia alba (acesso BGEN 09) e Lippia origanoides (acessos oro, ors e orv). Allium cepa (cebola) foi utilizado como modelo de avaliação de citogenotoxidade. As raízes de cebola foram expostas por 24 horas em diferentes concentrações de óleo essencial (diluído em 2,5% de acetona) (50, 100, 200, 300 e 400μg/mL). Como controle negativo foi utilizado água destilada e acetona 2,5%. E como controle positivo o Metil Metano Sulfonato (4x10-4M). Para ao preparo das lâminas a técnica utilizada foi o esmagamento. Parâmetros de ciclo celular e indução de alterações cromossômicas foram avaliados. Os óleos obtidos foram analisados por Cromatografia com fase gasosa acoplada com espectrometria de Massa (CG-EM). Os constituintes majoritários dos óleos essenciais foram: citral, trans-hidrato sabineno, 1,8-cineol e carvacrol para Lippia alba, L. origanoides, acessos oro, ors e orv respectivamente. De um modo geral, todos os óleos investigados interferiram na dinâmica normal do ciclo celular (interferência sobre índices mitóticos ou índices de fases). Estes efeitos foram mais evidentes para o óleo extraído de L. alba. Do mesmo modo, a exposição aos óleos essenciais evidenciou o aumento nos percentuais de algumas alterações cromossômicas, como cromossomos aderentes, perda de cromossomos, pontes cromossômicas e segregação tardia. Dentre os resultados observados, o envolvimento de alterações aneugênicas parece ser o efeito mais evidente. Possíveis relações entre as substâncias encontrados nos óleos destas espécies com as alterações observadas são discutidas. Este estudo sugere, portanto, o uso racional de plantas da espécie de Lippia para fins terapêuticos e mostra a necessidade de mais estudos sobre seu potencial citogenotóxico. / Many species of the genus Lippia (Verbenaceae family) characterized by their medicinal properties. Among the chemical and pharmacological aspects investigated in Lippia, those related to its essential oils stand out. Essential oils are produced by secondary metabolism and are complex mixtures of volatile and lipophilic substances. The aim of this study was to investigate the cytogenotoxic effects of essential oils from Lippia alba (BGEN 09 acess) and Lippia origanoides ( oro, ors and orv access). Allium cepa (onion) was used as a model for cytogenotoxicity evaluating. The onion roots were exposed for 24 hours at different concentrations of essential oil (diluted in 2.5% acetone) (50, 100, 200 , 300 and 400μg/mL). As a negative control was used distilled water and 2.5% acetone. And as the positive control Methyl methane sulfonate ( 4x10- 4M ). The slides were prepared by squashing. Parameters of cell cycle and induction of chromosomal alterations were evaluated. The oils were analyzed by Gas Chromatography - Mass Spectrometry (GC -MS). The major constituents of the essential oils were citral, trans - sabinene hydrate , 1,8- cineole and carvacrol for Lippia alba , L. origanoides (oro, ors and orv accesses) respectively. In general , all oils investigated interfered in the normal dynamics of the cell cycle (interference on mitotic index or phases indices). These effects were most evident for the oil extracted from L. alba. Similarly, exposure to the essential oils showed some increase in the percentage of chromosomal alterations, as adherent chromosomes, loss of chromosomes, later segregation and bridges. Among the observed results, the involvement of aneugenic changes seems to be the most obvious effect. Possible relationships between the compounds found in oils of these species with the changes observed are discussed. This study therefore suggests the rational use of plants of the species Lippia for therapeutic purposes and shows the need for more studies on its cytogenotoxic potential.

Orientador: Márcia Ortiz Mayo Marques / Resumo: A Lippia alba (Verbenaceae), popularmente conhecida como erva-cidreira, é uma planta originaria da América do Sul e considerada um recurso genético vegetal de altíssimo valor para estudos de bioprospecção pela sua potencial importância econômica, ecológica e propriedades do seu óleo essencial. A irrigação de sistemas de produção agrícola depende da disponibilidade dos recursos hídricos da demanda de água em cada etapa do cultivo, requerendo um manejo criterioso e sustentável. Estudos em plantas aromáticas têm mostrado que o manejo da água por meio da irrigação e as condições climáticas do local de cultivo são fatores abióticos determinantes na produção de biomassa, rendimento de óleo essencial e síntese de princípios ativos de interesse para a indústria de cosméticos, perfumes e fragrâncias. O objetivo deste estudo foi avaliar o efeito de diferentes lâminas de irrigação na biomassa, rendimento, produção e composição química do óleo essencial da espécie Lippia alba, de clones selecionados de quimiotipo linalol, pertencentes ao Banco de Germoplasma do Instituto Agronômico (IAC). As mudas foram obtidas por propagação assexuada e o cultivo conduzido em casa de vegetação em sistema orgânico de produção. Os tratamentos foram representados pelo manejo da irrigação com base na evapotranspiração de referência (50%, 75%, 100% ET0 e 125% da ET0). Os cortes foram efetuados aos 90, 180, 270 e 360 dias após a poda de formação (DAPF). As plantas foram avaliadas quanto à produção total de ma... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Lippia alba (Verbenaceae), popularly known as lemon balm, is a plant originally from South America and considered a highly valuable plant genetic resource for bioprospecting studies due to its potential economic, ecological and essential oil properties. The irrigation of agricultural production systems depends on the availability of water resources for water demand at each stage of cultivation, requiring careful and sustainable management. Studies on aromatic plants have shown that water management through irrigation and the climatic conditions of the place of cultivation are abiotic factors determining biomass production, essential oil yield and synthesis of active principles of interest to the cosmetics, perfumes industry and fragrances. The objective of this study was to evaluate the effect of different irrigation depths on biomass, yield, production and chemical composition of the essential oil of the species Lippia alba, from selected clones of the linalool chemotype, belonging to the Germplasm Bank of the Agronomic Institute (IAC). The seedlings were obtained by asexual propagation and cultivation was carried out in a greenhouse in an organic production system. The treatments were represented by irrigation management based on reference evapotranspiration (50%, 75%, 100% ET0 and 125% ET0). The cuts were made at 90, 180, 270 and 360 days after the formation pruning (DAPF). Plants were evaluated for total leaf dry matter production (PTMSF), total stem dry matter production... (Complete abstract click electronic access below) / Doutor

MSc (Chemistry) / Department of Chemistry / Malaria, an infectious disease affecting both human beings and other animals, is transmitted by parasitic protozoans belonging to the Plasmodium genus. Malaria is commonly treated with drugs such as quinine, chloroquine, and artesunate. However, the incidence of treatment failure due to drug-drug interactions and parasite resistance is increasing. Therefore, the rich medicinal potential of plants found in nature in Africa is increasingly being explored. The traditional use of Lippia javanica, Sclerocarya birrea, Melia azedarach and Capparis tomentosa for the treatment of malaria is well-known, but the phytochemistry of these four plants is not fully known. Parts of these plants were extracted and column chromatography was used to fractionate the extracts. The antioxidant activities of the fractions were determined using free radical scavenging and reducing power assays, while the cytotoxic, antiplasmodial and antitrypanosomal activities were determined using cell toxicity assay, parasite lactate dehydrogenase (pLDH) and trypanosome assay. The methanol stem bark extract of Melia azedarach (Fraction 2) had the highest phenolic content (59.39 mg GAE/g), while the methanol leaf extract of Melia azedarach had the highest flavonoid content of 188.65 mg QE/g. In the reducing power tests and DPPH free radical scavenging activity, the methanol stem bark extract of Melia azedarach had the lowest IC50 value of 0.1074 μg/mL and an IC0.5 value of 0.5296 μg/mL, respectively. Furthermore, the methanol stem bark extract of Melia azedarach at a concentration of 50 μg/mL showed significant cytotoxicity against HeLa cells (-1.22±0.07 %). The methanol stem bark extract of Melia azedarach at the tested concentration (250 μg/mL) decreased the viability of Plasmodium falciparum to 36.38±11.96 % with an IC50 value of 6.5 μg/mL. Concerning the antitrypanosomal activity, the methanol stem bark extract of Melia azedarach affected the viability of the trypanosomes at the tested concentration (250 μg/mL), giving a viability of 14.05 ± 0.59 %, with an IC50 value of 0.4 μg/mL. The presence of epicatechin (29) and catechin (31) in this extract was confirmed using several spectroscopic techniques (IR, NMR, UPLC-MS and HRMS). / NRF

O potencial farmacológico de espécies do gênero Lippia no tratamento de infecções é conhecido popularmente, assim foi considerado o estudo da atividade da Lippia salviaefolia nativa do cerrado brasileiro. O objetivo do trabalho foi avaliar a atividade antimicrobiana de extratos de Lippia salviaefolia Cham. e sua citotoxicidade, visando aplicação como conservante natural em formulações cosméticas. O ensaio antimicrobiano foi realizado através do método de difusão em placa e determinação do CMI por de microdiluição em microplacas, utilizando os microrganismos: P. aeruginosa ATCC 9027, S. aureus ATCC 6538, E. coli ATCC 8739, C. albicans ATCC 10231. Desenvolveu-se e avaliou-se a estabilidade acelerada de cremes, geis e xampus contendo o extrato de Lippia salviaefolia. As formulações de melhor desempenho quanto à estabilidade foram submetidas ao teste de eficácia de conservante. Avaliou-se segurança das formulações in vitro, em substituto cutâneo dermo-epidérmico por meio de estudos histológicos e o teste de citotoxicidade em cultura de queratinócitos humanos. No teste de atividade antimicrobiana verificou-se que a fração de acetato de etila e a fração de clorofórmio foram frações mais ativas com CMI de 0,2 % para bactérias e fungos. Os testes de eficácia de conservante das formulações contendo fr. ac. etila de L. salviaefolia apresentaram atividade antimicrobiana para bactérias e C. albicans em creme, gel e xampu, para A. niger apresentaram atividade adequada somente em xampu. No teste de citoxicidade foi verificado segurança em concentrações a partir de 0,05 %. Sendo que na concentração de 0,2 %, correspondente ao CMI, se atinge a faixa de segurança apenas com meia hora de contato. Os resultados sugerem que a fração de acetato de etila da Lippia salviaefolia Cham. pode ser utilizada como conservante natural em produtos cosméticos. / The pharmacological potential of species of the Lippia in the treatment of infections has been known popularly, thus was considered the study of the activity of the native salviaefolia Lippia of the Brazilian Cerrado. The purpose of the work was to evaluate the antimicrobial activity of extracts of Lippia salviaefolia Cham. and its citotoxicy aiming the application as natural preservative in cosmetic formulations. The assay antimicrobial was carried through the method of diffusion in plate and determination of the MIC by microdilution, having used the microorganisms: P.aeruginosa ATCC 9027, S. aureus ATCC 6538, E. coli ATCC 8739, C. albicans ATCC 10231. It has developed and evaluated the stability of creams, gels and xampus with the extract of Lippia salviaefolia Cham. The formulations of the best stability performance have submitted to preservative efficacy test. The safe of the formulations has histological evaluated by in vitro. In the test of antimicrobial activity it has verified that the ethyl acetate and the chloroform fraction have been the most active and with CMI of 0.2 % for bacteria and fungus. The tests of effectiveness of preservatives have showed of the formulations contend the ethyl acetate fraction presented antimicrobial activity for bacteria and C. albican in cream, gel and shampoo. On the other hand, against A.niger it has been suitable activity in shampoo. In the citotoxity test, the 0,05 % ethyl acetate fraction has presented interval of security. And in the concentration of 0,2%, corresponding to CMI, safety\'s strip is just reached with half hour of contact. The results have suggested that the ethyl acetate extract of the Lippia salviaefolia Cham. should be used as natural preservative in cosmetic products.

Submitted by Maike Costa (maiksebas@gmail.com) on 2017-09-12T13:51:31Z No. of bitstreams: 1 arquivototal.pdf: 2565770 bytes, checksum: 9a17a1e982b430cc6e63bdb785f2ce92 (MD5) / Made available in DSpace on 2017-09-12T13:51:31Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 2565770 bytes, checksum: 9a17a1e982b430cc6e63bdb785f2ce92 (MD5) Previous issue date: 2015-03-31 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Lippia microphylla Cham. (Verbenaceae), known as “alecrim-do-mato”, “alecrim-de-tabuleiro” and “alecrim-pimenta”, is populary used as antiseptic or to treat respiratory disorders. From its leaves, the essential oil (LM-OE) was extracted, that, according to Silva (2013), presented tocolytic effect on rat, with a probably indirect inhibition of Ca2+ influx through voltage-gated calcium channels (CaV). Thus, the aim of this work was to characterize the tocolytic action mechanism of LM-OE on rat. Isometric contractions were recorded to determine and compare the relative efficacy and potency (n = 5). Considering that the NO pathway can negatively modulate the CaV, we decided to evaluate this participation on rat uterus pre-contracted by oxytocin. For that, it was employed L-NAME, a non-selective inhibitor of NO synthase (NOS). The results show that the tocolytic potency of LM-OE (EC50 = 2.7 ± 0.6 μg/mL) was approximately 9.5-fold attenuated in the presence of L-NAME (EC50 = 25.6 ± 3.9 μg/mL). Furthermore, this reduction was reversed in the presence of the L-arginine (EC50 = 6.2 ± 1.7 μg/mL), a NOS substrate, and simultaneous presence of the L-arginine and L-NAME (EC50 = 7.2 ± 1.4 μg/mL), confirming the participation of NO pathway in the tocolytic action of LM-OE. Similarly, the relaxant potency of LM-OE was approximately 3.7-fold reduced in the presence of ODQ (EC50 = 10.1 ± 0.9 μg/mL), a soluble guanylate cyclase (sGC) inhibitor, and 3.3-fold in the presence of Rp-8-Br-PET-cGMPS (EC50 = 8.9 ± 0.9 μg/mL), a protein kinase G (PKG) inhibitor. Thus, LM-OE positively modulates the NO/sGC/PKG pathway to promote its tocolytic effect on rat. Given that this pathway can activate the K+ channels, it was questioned if these channels would be involved on tocolytic effect of LM-OE. To confirm this hypothesis, experiments were made in the presence of CsCl, a non-selective K+ channels blocker, and it was observed that the concentration-response curve of LM-OE (EC50 = 2.7 ± 0.6 μg/mL) was rightward shifted with an approximately 4.3-fold reduction on its relaxant potency, confirming the participation of K+ channels on tocolytic effect of essential oil. In order to verify the subtypes involved, it was employed selective blockers of K+ channels. Control concentration-response curve was also shifted to the right and the tocolytic potency of LM-OE was attenuated in the presence of tetraethylammonium (EC50 = 10.5 ± 0.3 μg/mL), apamin (EC50 = 10.0 ± 1.0 μg/mL), 4-aminopyridine (EC50 = 12.4 ± 2.0 μg/mL) and glibenclamide (EC50 = 9.7 ± 0.9 μg/mL), the large (BKCa) and small conductance (SKCa) calcium-activated, voltage-gated (KV) and adenosine triphosphate-sensitive (KATP) potassium channels blockers, respectively. Moreover, LM-OE had its tocolytic potency reduced in the same intensity when these four blockers were simultaneously incubated (EC50 = 16.6 ± 2.6 μg/mL), not observing synergistic effect. Therefore, the tocolytic action mechanism of LM-OE on rat involves the positive modulation of the NO/sGC/PKG/K+ channels pathway, suggesting the blockade of calcium influx through CaV, leading to the relaxation of uterine smooth muscle. / Lippia microphylla Cham. (Verbenaceae), conhecida como “alecrim-do-mato”, “alecrim-de-tabuleiro” e “alecrim-pimenta”, é usada popularmente como antisséptico ou para tratar distúrbios respiratórios. Das folhas dessa espécie foi extraído o óleo essencial (LM-OE) que, de acordo com Silva (2013), apresentou efeito tocolítico em útero isolado de rata, indicando uma provável inibição indireta do influxo de Ca2+ através dos canais de cálcio dependentes de voltagem (CaV). Diante disso, o objetivo deste trabalho foi caracterizar o mecanismo de ação tocolítica do LM-OE em rata. As contrações isométricas foram monitoradas para determinar e comparar a eficácia e a potência relativas (n = 5). Tendo em vista que os CaV podem ser modulados negativamente pela via do óxido nítrico (NO), decidiu-se avaliar a participação dessa via em útero de rata pré-contraído com ocitocina. Para tanto, utilizou-se L-NAME, um inibidor não seletivo de sintase do NO (NOS). Os resultados evidenciaram que a potência tocolítica do LM-OE (CE50 = 2,7 ± 0,6 μg/mL) foi reduzida em aproximadamente 9,5 vezes, na presença de L-NAME (CE50 = 25,6 ± 3,9 μg/mL), sendo essa redução revertida na presença de L-arginina (CE50 = 6,2 ± 1,7 μg/mL), substrato para a NOS, e na presença simultânea de L-arginina e de L-NAME (CE50 = 7,2 ± 1,4 μg/mL), confirmando o envolvimento da via do NO na ação tocolítica do LM-OE. Semelhantemente, a potência relaxante do LM-OE foi reduzida em aproximadamente 3,7 vezes na presença de ODQ (CE50 = 10,1 ± 0,9 μg/mL), inibidor de ciclase de guanilil solúvel (sGC), e 3,3 vezes na presença de Rp-8-Br-PET-cGMPS (CE50 = 8,9 ± 0,9 μg/mL), inibidor de proteína cinase G (PKG). Sendo assim, o LM-OE modula positivamente a via do NO/sCG/PKG para promover seu efeito tocolítico em rata. Uma vez que essa via pode ativar canais de K+, questionou-se se esses canais estariam participando do efeito tocolítico do LM-OE. Para confirmar essa hipótese, foram realizados experimentos na presença de CsCl, bloqueador não seletivo de canais de K+, e observou-se que a curva concentração-resposta do LM-OE (CE50 = 2,7 ± 0,6 μg/mL) foi desviada para a direita com redução da potência relaxante (CE50 = 11,7 ± 1,5 μg/mL) em aproximadamente 4,3 vezes, confirmando a participação dos canais de K+ no efeito tocolítico do óleo essencial. Para verificar quais seriam esses subtipos de canais utilizou-se bloqueadores seletivos. A curva controle concentração-resposta também foi desviada para a direita e a potência tocolítica reduzida na presença de tetraetilamônio (CE50 = 10,5 ± 0,3 μg/mL), de apamina (CE50 = 10,0 ± 1,0 μg/mL), de 4-aminopiridina (CE50 = 12,4 ± 2,0 μg/mL) e de glibenclamida (CE50 = 9,7 ± 0,9 μg/mL), bloqueadores de canais de potássio: ativados por cálcio de grande (BKCa) e de pequena condutância (SKCa), dependentes de voltagem (KV) e sensíveis ao trifosfato de adenosina (KATP), respectivamente. Ademais, o LM-OE teve sua potência tocolítica reduzida quando esses quatro bloqueadores foram pré-incubados simultaneamente (CE50 = 16,6 ± 2,6 μg/mL), não sendo observado efeito sinérgico. Assim, o mecanismo de ação tocolítica do LM-OE em rata envolve a modulação positiva da via NO/sGC/PKG/canais de K+, sugerindo o bloqueio do influxo de cálcio via CaV e, consequentemente, o relaxamento do músculo liso uterino.