Angélica Maria Lucchese

β‐Cyclodextrin Complex Containing Lippia grata Leaf Essential Oil Reduces Orofacial Nociception in Mice – Evidence of Possible Involvement of Descending Inhibitory Pain Modulation Pathway

The treatment of orofacial pain remains a major challenge for modern medicine. Thus, we prepared and physicochemically characterized a new β‐cyclodextrin complex containing Lippia grata leaf essential oil (β‐CD/EO) to investigate their possible antinociceptive activity in animal models of orofacial pain. The results of Differential scanning calorimeter (DSC) and Thermogravimetry/derivative thermogravimetry (TG/DTG) showed that the products prepared by Slurry complexation (SC) method were able to incorporate greater amounts of EO. In the X‐ray diffractogram, it was shown that complex between EO and β‐CD was formed. Male Swiss mice were pre‐treated with β‐CD/EO (6, 12 or 24 mg/kg, per os, gavage, p.o.), morphine (5 mg/kg, i.p.) or vehicle (distilled water, p.o.) 1 hr before treatment with formalin (20 μL, 2%), capsaicin (20 μL, 2.5 μg) or glutamate (40 μL, 25 μM) into the right upper lip. Our results demonstrated that p.o. treatment with β‐CD/EO was significantly (p < 0.05 or p < 0.001) capable of reducing the nociceptive face‐rubbing behaviour in both phases of the formalin test. β‐CD/EO‐treated mice were also significantly (p < 0.05 or p < 0.001) protected against nociception induced by capsaicin and glutamate. For the action in the central nervous system (CNS), ninety minutes after the treatment, the mice were perfused, the brains collected, crioprotected, cut in a criostate and submitted to an immunofluorescence protocol for Fos protein. The immunofluorescence protocol demonstrated that the β‐CD/EO significantly activated (p < 0.05; p < 0.01 or p < 0.001) the motor cortex, the Locus ceruleus, the nucleus raphe magnus and the periaqueductal gray of the CNS. These effects apparently did not alter, in tested doses, the motor coordination of mice in the rota‐rod test. Our results proposed that β‐CD/EO might present an important draft of drug to the study of new compounds for the treatment of orofacial pain.

Inclusion of terpenes in cyclodextrins: Preparation, characterization and pharmacological approaches

Terpenes constitute the largest class of natural products and are important resources for the pharmaceutical, food and cosmetics industries. However, due to their low water solubility and poor bioavailability there has been a search for compounds that could improve their physicochemical properties. Cyclodextrins (natural and derived) have been proposed for this role and have been complexed with different types of terpenes. This complexation has been demonstrated by using analytical techniques for characterizing complexes such as DSC, NMR, XRD, FTIR, and TGA. The formation of inclusion complexes has been able to improve drug characteristics such as bioavailability, solubility and stability; and to enhance biological activity and efficacy. This review shows strong experimental evidence that cyclodextrins improve the pharmacological properties of terpenes, and therefore need to be recognized as being possible targets for clinical use.

Antiinflammatory and antinociceptive activities of Blechnum occidentale L. extract.

AIM OF STUDY Blechnum occidentale L. is a terrestrial fern that ranges from the United States to South America, and is employed in Brazilian folk medicine. In the present study we investigated the antinociceptive and antiinflammatory activities of the methanolic extract of Blechnum occidentale L. (MEB) in animal models of pain and inflammation to support its medicinal use in treatment of inflammatory and pulmonary diseases, urinary infections and liver diseases. MATERIALS AND METHODS The antinociceptive activity of MEB was evaluated using the writhing, formalin, and tail flick tests. The antiinflammatory activity of MEB was evaluated in carrageenan-induced paw oedema and neutrophil migration. In order to discard possible non-specific muscle relaxant or sedative effects of MEB, mice motor performance was evaluated in the rota rod test and its toxicity evaluated over 14 days. RESULTS Intraperitoneal (IP) administration of MEB (0.01-100mg/kg) produced a dose-related antinociception on acetic acid-induced writhing in mice. Oral administration of MEB, at a different range of doses (100-400 mg/kg), also produced significant antinociceptive effect on the writhing test. Furthermore, treatment with MEB (100 and 200 mg/kg IP) inhibited significantly both the early and late phases of formalin-induced hypernociception in rats. In contrast, treatment with MEB (100 and 200 mg/kg IP) did not prevent the thermal nociception in the tail flick test. The IP administration of MEB (100 and 300 mg/kg) significantly reduced the paw oedema induced by carrageenan. Moreover, systemic treatment with MEB (11-300 mg/kg) reduced the neutrophil migration in the carrageenan-induced migration to the peritoneal cavity. In the rota rod test, MEB-treated mice did not show any significant motor performance alterations with the dose of 300 mg/kg. In addition, over the study duration of 14 days, there were no deaths or toxic signs recorded in the mice given 100 or 1000 mg/kg of MEB. CONCLUSION The results described here are the first report of pharmacological studies of Blechnum occidentale L. and indicate that this plant has antinociceptive and antiinflammatory activities which support its folk medicine use.

Propagação vegetativa de Hyptis leucocephala Mart. ex Benth. e Hyptis platanifolia Mart. ex Benth. (Lamiaceae)

The genus Hyptis (Lamiaceae) has a special metabolism with remarkable variability and predominance of essential oils of great value to the various communities that use these plants due to their therapeutic properties. A number of studies on this genus have demonstrated that its essential oils present antimicrobial, antifungal, cytotoxic, anti-inflammatory, anti-HIV and insecticidal activities. However, few studies have focused on the sustainable exploitation of these species. The aim of this study was to evaluate the effect of different types of cuttings and different concentrations of indolebutyric acid (IBA) on the survival, growth and rooting rates of cuttings from Hyptis leucocephala Mart. ex Benth. and Hyptis platanifolia Mart. ex Benth., species endemic to the semi-arid region of Northeastern Brazil and with largely proven medicinal value. The experiment lasted four months. Apical, middle-apical, middle-basal and basal cuttings were treated with 0, 1000, 2000 and 4000 mg L-1 IBA solutions for 5 min. The use of apical, middle-apical and middle-basal cuttings induced by 2000 and 4000 mg L-1 IBA led to the best results for vegetative propagation of H. leucocephala. For H. platanifolia, the best results were obtained using middle-basal and basal cuttings treated with IBA at 2000 and 4000 mg L-1.

Antinociceptive and antiinflammatory activities of Adiantum latifolium Lam.: Evidence for a role of IL-1β inhibition

AIM OF STUDY Adiantum, one of the most widely distributed genera of the family Pteridaceae, is employed in folk medicine worldwide. Adiantum latifolium Lam. has been used in Latin American traditional medicine as anxiolytic, analgesic and antiinflammatory. The present study investigates the antinociceptive and antiinflammatory properties of the methanolic extract of Adiantum latifolium (MEA) in animal models of pain and inflammation to confirm its medicinal use. MATERIAL AND METHODS The antinociceptive and antiinflammatory activities of MEA were evaluated using the writhing, formalin, and tail-flick tests, carrageenan-induced paw edema and arachidonic acid-induced ear edema. Mice motor performance was evaluated in the rota rod test and the acute toxicity evaluated over 14 days. RESULTS Intraperitoneal (1-100mg/kg) or oral (100-400mg/kg) administration of MEA produced a dose-related inhibition of acetic acid-induced writhing in mouse. Furthermore, treatment with MEA (100mg/kg) inhibited both the early and late phases of formalin-induced hypernociception. In contrast, MEA (100mg/kg/IP) did not prevent the thermal nociception in the tail-flick test. In addition, MEA (100 and 200mg/kg/IP) inhibited important events related to the inflammatory response induced by carrageenan or arachidonic acid, namely local edema and increase in tissue interleukin-1β levels. MEA (300mg/kg/IP)-treated mice did not show any motor performance alterations. Over the study period of 14 days, there were no deaths or toxic signs recorded in the group of mice given 1000mg/kg of MEA. CONCLUSION The results demonstrate that Adiantum latifolium has antinociceptive and antiinflammatory activities, acting through the inhibition of IL-1β production.

Atividade antimicrobiana de méis de cinco espécies de abelhas brasileiras sem ferrão

The antimicrobial activity of honey produced by Melipona asilvai, Melipona quadrifasciata anthidioides, Friseomelita doederleinei, Tetragonisca angustula and Plebeia sp. were investigated. The agar well diffusion assay demonstrated that all honeys had antibacterial activity against Staphylococcus aureus, but only the samples from M. quadrifasciata anthidioides and F. doederleinei inhibited the growth of Escherichia coli. In the Minimum Inhibitory Concentration determination assay, M. asilvai, M. quadrifasciata anthidioides, F. doederleinei and T. angustula honeys were more active than that from Plebeia sp. for S. aureus and E. coli. The microorganisms Pseudomonas aeruginosa and Candida albicans were resistant to the all native stingless bee honeys in both assays. Honeys were more effective against bacteria than a sugar solution, suggesting that the mechanism for bacterial growth inhibition is not only related to the osmotic effect. The results of antimicrobial activity may explain the popular medicinal use of these honeys in bacterial diseases.

Variação no número de glândulas e produção de óleo em flores de Stigmaphyllon paralias A. Juss. (Malpighiaceae)

A population of Stigmaphyllon paralias (Malpighiaceae) was studied in Feira de Santana, Bahia, Brazil. Variation of the number of oil glands and oil production in the flowers were analized. 76% of the individuals did not present variation on the number of glands among their flowers, 41% presented flowers with 10 glands, 31% with eight glands and 4% with nine glands. In the flowers of the morphs with eight or nine glands, the absent glands were always associated to the inferior sepal. Only Centris leprieuri and Epicharis sp. (Apidae: Centridini) visited flowers of S. paralias, collecting oil as rewarding. The inferior sepal is located under the pollinator abdomen when it is standing in the flower, therefore being inaccessible for oil collection. We suggest that the occurrence of glands not utilized by pollinators has allowed the appearance of morphs in the populations that do not present those glands. All glands of all flower morphs are functional. Flowers of the two more representative morphs, bearing eight and 10 glands, produce the same amount of oil. Glands of flowers with lower number of glands (eight) increase nearly 20% plus oil production, compensating oil production per flower.

Chemical composition and pharmacological properties of the essential oils obtained seasonally from Lippia thymoides.

Abstract Context: Lippia thymoides Mart. & Schauer (Verbenaceae) is used in folk medicine to treat wounds, fever, bronchitis, rheumatism, headaches, and weakness. Objective: This study determinates the chemical composition of essential oils from L. thymoides, obtained at during each of the four seasons and correlates with pharmacological properties. Materials and methods: Essential oils were obtained by hydrodistillation and analyzed by gas chromatography coupled to mass spectroscopy (GC-MS). Antioxidant activity was determined by DPPH free radical scavenging and β-carotene bleaching methods. The antimicrobial assays were performed by minimum inhibitory concentration (MIC) and minimum microbicidal concentration (MMC) methods. Isolated rat aorta and uterus, and guinea-pig trachea were utilized to evaluate relaxant potential in pre-contracted smooth muscle. Results and discussion: Essential oils from leaves of L. thymoides had the sesquiterpene β-caryophyllene (17.22–26.27%) as the major constituent followed by borneol (4.45–7.36%), camphor (3.22–8.61%), camphene (2.64–5.66%), and germacrene D (4.72–6.18%). In vitro assays showed that these essential oils do not have antioxidant activity, have antimicrobial selectivity to Gram-positive bacteria Staphylococcus aureus (MIC = 0.004 mg/mL and MMC = 0.26–10.19 mg/mL) and Micrococcus luteus (MIC = 0.03 mg/mL and MMC = 8.43 mg/mL), relax isolated rat aorta (EC50 = 305–544 μg/mL, with endothelium; and EC50 = 150–283 μg/mL, without endothelium), and uterus (EC50 = 74–257 μg/mL), and minor potency, isolated guinea-pig trachea. Conclusions: Lippia thymoides is a source of natural products of pharmaceutical interest, being necessary additional studies to determine the substances involved in the biological activities.

Antimicrobial Activity of Lippia Species from the Brazilian Semiarid Region Traditionally Used as Antiseptic and Anti-Infective Agents

Lippia origanoides Kunth, Lippia alnifolia Schauer, and Lippia thymoides Martius and Schauer are shrubs used in the traditional Brazilian medicine as antiseptics, as well as in the treatment of infectious diseases. This study was designed to investigate the antibacterial and antifungal activities of the methanolic extracts of these species, as new potential sources of antimicrobial drugs. The antimicrobial activity of methanolic extracts was investigated against resistant yeasts and bacteria by agar disk diffusion. Then, the MIC determination of the most active species and its fractions in hexane, dichloromethane, ethyl acetate, and water was performed. By the agar diffusion assay, all species were active against at least two microorganisms, giving evidence to support their use in the popular medicine. L. origanoides leaves exhibited the widest antimicrobial action, inhibiting the growth of two Gram-positive bacteria and two yeasts; this activity was also confirmed by the MIC evaluation. The fractionation of L. origanoides crude extracts improved the activity in spectrum and intensity. The results obtained in this study indicate that L. origanoides may be a promising alternative in the treatment of bacterial and fungal infections and in the seeking of new antimicrobial drugs.

Antimicrobial activity of Syagrus coronata (Martius) Beccari

ABSTRACT This work aimed to evaluate the antimicrobial activity of aqueous and methanol extracts of leaves, inflorescences, nut-shell, liquid and solid endosperm nuts of Syagrus coronata against pathogenic bacteria and yeast. Screening was initially performed using the agar dilution method. The extracts regarded as bioactive underwent liquid-liquid partition for determination of their minimum inhibitory concentration and minimum bactericide concentration (MIC and MBC) and those of their respective fractions against the microorganisms inhibited in preliminary tests. Antimicrobial activity was observed only in inflorescences. The corresponding aqueous extract was effective against B. cereus and the three strains of S. aureus , and the corresponding MIC and MCB values were lower than those of dichloromethane, ethyl acetate and butanol fractions of the same extract. The methanol extract was effective against B. cereus , and the corresponding MIC and MBC values were higher than those of ethyl acetate and butanol fractions of the same extract. Key words: Syagrus coronata , Arecaceae, antimicrobial activity