Anti-inflammatory, antinociceptive and antioxidant activities of the hydromethanolic fraction from Annona nutans leaves / Atividade anti-inflamatória, antinociceptiva e antioxidante da fração hidrometanólica das folhas de annona nutans

Annona nutans (Annonaceae) is a plant species found in Bolivia, Paraguay, Argentina, and the Brazilian Cerrado, specifically in the states of Mato Grosso and Mato Grosso do Sul (Brazil). Its common names are Araticû-Mi and Araticû-Ñu. The research contributions regarding the chemical composition and biological activities of extracts from A. nutans are rare, with only four articles being found in the literature. Therefore, the present study evaluated the anti-inflammatory and antinociceptive activities of the hydromethanolic fraction (FHMeOH) using carrageenan-induced paw edema and hot-plate tests. In addition, the antioxidant activity was evaluated by DPPH radical scavenging, total phenolic, flavonoid and tannin content assays and quantification of the major metabolites by LC-MS were performed. Oral treatment with the FHMeOH (at a dose of 300 mg.kg-1) significantly reduced paw edema 2 h and 4 h after the inflammatory stimulus. The intraperitoneal (i.p.) treatment with the FHMeOH (50 and 100 mg.kg-1) proved to be most effective, and the inhibition of acute inflammation was still visible 6 h after carrageenan injection. At doses of 50 and 100 mg.kg-1 (i.p.), FHMeOH exhibits central antinociceptive effects by increasing the latency of the reaction in the hot-plate model. The FHMeOH showed antioxidant potential, and the metabolites quercetin-3-O-galactoside, quercetin-3-O-glucoside, isorhamnetin-3-O-galactoside, quercetin-3-O-ß-D-apiofuranosyl-(1→2)-galactopyranoside, and chlorogenic acid were identified and quantified by LC-MS. Our results indicate, for the first time, that FHMeOH has anti-inflammatory, antinociceptive and antioxidant potential, and it is a promising source of studies for new herbal medicines

Annona nutans (Annonaceae) é uma espécie de planta encontrada na Bolívia, Paraguai, Argentina e no Cerrado brasileiro, especificamente nos estados de Mato Grosso e Mato Grosso do Sul (Brasil). Seus nomes mais comuns são aratico e Araticû-Mi e Araticû-Ñu. As contribuições da pesquisa em relação à composição química e atividades biológicas dos extratos de A. nutans são raras, com apenas quatro artigos encontrados na literatura. Portanto, o presente estudo avaliou as atividades anti-inflamatória e antinociceptiva da fração hidrometanólica (FHMeOH) utilizando edema de pata induzido por carragenina e testes de placaquente. Além disso, a atividade antioxidante foi avaliada por meio de sequestro de radical DPPH, e foram realizados ensaios de quantificação de fenóis, flavonoides e taninos totais e quantificação dos principais metabólitos por CL-EM. O tratamento oral com a FHMeOH (na dose de 300 mg.kg-1) reduziu significativamente o edema da pata 2 e 4 h após o estímulo inflamatório. Por outro lado, o tratamento intraperitoneal (i.p.) com FHMeOH (50 e 100 mg.kg-1) provou ser mais eficaz e a inibição da inflamação aguda foi ainda visível 6 horas após a injeção de carragenina. Nas doses de 50 e 100 mg.kg-1 (i.p.), FHMeOH exibiu efeitos antinociceptivos centrais aumentando a latência da reação no modelo de placa quente. FHMeOH apresentou potencial antioxidante e os metabólitos quercetina-3-O-galactosídeo, quercetina-3-O-glicosídeo, isoramnetina-3-O-galactosídeo, quercetina- 3-O-ß-D-apiofuranosil-(1 → 2)-galactopiranosídeo e ácido clorogênico foram identificados e quantificados por CL-EM. Nossos resultados indicam, pela primeira vez, que o FHMeOH possui efeitos anti-inflamatórios, antinociceptivos e antioxidantes, sendo uma fonte promissora de estudos para novos medicamentos fitoterápicos.

Chemical composition and evaluation of the anti-inflammatory and antinociceptive activities of Duguetia furfuracea essential oil: Effect on edema, leukocyte recruitment, tumor necrosis factor alpha production, iNOS expression, and adenosinergic and opioidergic systems.

ETHNOPHARMACOLOGICAL RELEVANCE: Duguetia furfuracea (A. St. -Hil.) Saff. (Annonaceae) is commonly known in Brazil as "araticum-seco," and its root is used in folk medicine to treat inflammatory and painful disorders. However, no studies have been performed to evaluate these therapeutic activities. AIM OF THE STUDY: Investigate the chemical composition, anti-inflammatory and antinociceptive effects, and elucidate the possible antinociceptive mechanisms of action from the essential oil of D. furfuracea (EODf) underground stem bark. MATERIALS AND METHODS: Chemical composition was determined by gas chromatography and mass spectrometry (GC/MS). The paw edema induced by LPS, formalin-induced nociception, LPS-induced thermal hyperalgesia and rota-rod tests in vivo were used to evaluate the anti-inflammatory and antinociceptive effects in addition to the alteration on motor coordination. Histological analyses and an immunohistochemistry assay for iNOS were performed on mouse footpads of naive, control, 10 mg/kg EODf, and 10 mg/kg indomethacin (Ind) groups. The samples were removed at 1, 3, and 6 h after subplantar injection of LPS. In addition, the involvement of the adenosinergic, opioidergic, serotonergic, and cholinergic systems were investigated, in order to elucidate possible antinociceptive mechanisms. RESULTS: Twenty-four volatile constituents were detected and identified. (E)-asarone (21.9%), bicyclogermacrene (16.7%), 2,4,5-trimethoxystyrene (16.1%), α-gurjunene (15%), cyperene (7.8%), and (E)-caryophyllene (4.6%) were major compounds found in EODf. Oral treatment (p.o.) with EODf (1, 3, and 10 mg/kg) significantly inhibited the paw edema induced by LPS. At 10 mg/kg EODf promoted inhibition of tumor necrosis factor alpha (TNF-α) production, recruitment of polymorphonuclear (PMN) leukocytes and inducible nitric oxide synthase (iNOS) expression in paw tissue. EODf (10 and 30 mg/kg, p.o.) also reduced licking time in both phases of the formalin test and it had a significant effect on the LPS-induced thermal hyperalgesia model. The administration of caffeine (Caf) and naloxone (Nal) reversed the antinociceptive activity of EODf, in the first phase of the formalin test and in the LPS-induced thermal hyperalgesia model. Moreover, Nal was also able to abolish the antinociception caused by EODf, in the second phase of formalin test. In the rota-rod test, EODf-treated animals did not show any alteration of motor coordination. CONCLUSIONS: Our findings indicate that EODf underground stem bark produces anti-inflammatory and both central and peripheral antinociceptive effects. Furthermore, the antinociceptive activity of EODf underground stem bark is possibly mediated by adenosinergic and opioidergic pathways, and its properties do not induce effects on motor coordination. These results support the use of the folk medicine, D. furfuracea root, to treat inflammation and painful conditions.

Maternal exposure to Cochlospermum regium: a toxicological evaluation

Cochlospermum regium (Schrank) Pilg., Bixaceae, is a Brazilian plant widely used as a folk medicine in the southwestern of the Brazil to treat inflammation and infection diseases. However, the effects of C. regium hydroethanolic extract on pregnant rats have not been assessed. To evaluate the effects of the C. regium on pregnant rats during the organogenic period, the hydroethanolic extract was administered via gavage at a dose of 11.5 mg/kg/day to rats from 6th to 15th day of pregnancy. No clinical signs of maternal toxicity were observed. The placenta's and fetuses' weight were similar in control and treated animals. The term fetuses dis not present malformations or anomalies although the number of live fetuses and birth rate were significantly decreased. In conclusion, the C. regium hydroethanolic extract is nontoxicant to the pregnant rat although it would be likely to interfere in the progress of the embryofetal development.

Brine shrimp lethality test as a biological model for preliminary selection of pediculicidal components from natural source / Teste de toxicidade frente artemia como modelo preliminar de busca de substâncias pediculicidas a partir de fontes naturais

Brine shrimp lethality test as a biological model for the preliminary selection of pediculicidal components from a natural source. In order to achieve a good correlation between pediculicidal activity and brine shrimp lethality (BSL) test, several pediculicidal substances and active essential oils were tested in BSL test, with the purpose to use the latter as convenient preliminary protocol for pediculicidal activity. Benzyl benzoate, deltametrine and essential oil of Eucalyptus were purchased and clove essential oil obtained by hydrodistillation, besides essential oils, chloroform extracts from Duguetia furfuracea were also submitted to BSL test. All of them were carried out with same protocol described to pediculicidal assay found in the literature, i.e, flask tests were examined every five minutes in the first half hour and then every ten minutes until all the naupli were dead or no movements were observed (knockdown). During the BSL test, it was possible to observe the effect of a particular lethal dose or only a knockdown in the arthropod, as occurred in the test with lice. The results of the BSL test for essential oils and other active substances are essentially in agreement with those described in literature for pediculicidal activity. Extracts and essential oil obtained from aerial parts of D. furfuracea did not present activity, but the essential oil from underground stem bark was active. α-asarone has already been isolated from the underground stem and it has been previously described to possess insecticidal activity.

O teste de toxicidade frente a Artemia salina (TAS) foi utilizado como modelo biológico preliminar na busca de substâncias potencialmente pediculicidas a partir de fontes naturais. Foi encontrada uma boa correlação entre a atividade pediculicida e o TAS, várias substâncias e óleos essenciais descritos como pediculicidas foram testados sobre o microcrustáceo, com o objetivo de se obter um protocolo preliminar e apropriado para detectar aquela atividade. Benzoato de benzila, deltametrina, e óleos essenciais de eucalipto e cravo foram obtidos comercialmente e/ou por extração. Além desses, extratos e óleos essenciais de Duguetia furfuracea também foram testados frente a Artemia. Todas as amostras foram conduzidas utilizando o mesmo protocolo proposto para atividade pediculicida, ou seja, os frascos testes foram lidos a cada cinco minutos na primeira meia hora e depois a cada dez minutos até os nauplios estarem mortos ou sem movimento (knockdown). Durante o TAS foi possível observar a dose letal ou somente o knockdown na larva, como ocorrido no teste com piolho. Os resultados do TAS para os óleos essencial e os demais compostos ativos estão essencialmente de acordo com o descrito na literatura para a atividade pediculicida. Extratos das sementes e óleos essenciais das folhas e sementes D. furfuracea não apresentaram atividade, mas o óleo das cascas do caule subterrâneo foi ativo α-asarona foi isolado das cascas do caule e esta substância apresenta atividade inseticida.