Antinociceptive and anti-inflammatory effect of Poincianella pyramidalis (Tul.) L.P. Queiroz.

ETHNOPHARMACOLOGY RELEVANCE: Poncianella pyramidalis (Leguminosae) is a Caatinga plant used in folk medicine because of its pharmacological properties, which include anti-inflammatory action. However, chemical compounds responsible for this effect have not yet been identified. AIM OF THE STUDY: This study aimed to evaluate the antioxidant, antinociceptive and anti-inflammatory effects of the ethyl acetate fraction from the inner bark of P. pyramidalis. MATERIAL AND METHODS: Total phenol content (TP) was estimated using the Folin-Ciocalteu reagent, while in vitro antioxidant activity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging assay. Chemical identification was done using LC-PDA/MS and LC-ESI/MS/MS. In vivo antinociceptive and anti-inflammatory properties were investigated using formalin, mechanical hypernociception and carrageenan-induced pleurisy assays in mice. RESULTS: TP was 525.08 ± 17.49 µg mg-1 gallic acid equivalent. The ethyl acetate fraction (EAF) inhibited 87.76% of the DPPH radical with an EC50 of 22.94 µg mL-1 and Antioxidant Activity Index of 1.74. LC-PDA/MS and LC-ESI/MS/MS identified 15 compounds that are mostly derived from gallic and ellagic acids. Regarding in vivo antinociceptive and anti-inflammatory activity, EAF (100 mg kg-1) significantly reduced the nociceptive response in the second phase of the formalin assay by 50% (p < 0.01) compared with the control group. In the hypernociception test, a significant (p < 0.001) anti-hyperalgesic effect of EAF (100 mg kg-1) was observed up to the third hour of evaluation (p < 0.001). In the carrageenan assay, EAF (100 mg kg-1) was shown to inhibit protein extravasation, increase total leukocytes and neutrophils, and inhibit mononuclear cells. CONCLUSION: These results demonstrate EAF from the inner bark of P. pyramidalis has strong in vitro antioxidant effect as well as in vivo antinociceptive and anti-inflammatory activities, which may be attributed to the bark being rich in phenolic compounds derived from gallic acid.

Hyptis pectinata gel prevents alveolar bone resorption in experimental periodontitis in rats

Hyptis pectinata (L.) Poit., Lamiaceae, is an aromatic, abundant and broadly used plant species in Sergipe to treat oral and gastrointestinal pain and inflammation. The aim of the present study was to analyze the relation between periodontitis and changes in the corporal mass and alveolar bone structure after induction of experimental periodontal disease in rat treated or not treated with H. pectinata gel at 5% (GS5%) and 10% (GS10%), comparing their effects with doxycycline gel at 10% (D10%, positive control), vehicle gel (negative control) and a group with experimental periodontal disease, but non-treated. The gels were locally applied in the gingival region immediately after the experimental periodontal disease induction by ligature (3×/day, 11 days). Bone destruction was determined through clinical exam, histopathological analysis and cone beam computed tomography of the experimental animals (n = 36). After 11 days of periodontitis induction, all groups that received ligature presented a decrease in the corporal mass, except to the naïve group (without experimental periodontal disease) (p < 0.05). Computed tomography results have shown healthy bone structure in the group I and bone resorption for the test groups. Histopathological analysis confirmed the healthy bone structure for naïve group animals, while the test groups exhibited bone loss in several degrees. In particular, the non-treated group animals had an intense inflammatory process. When the periodontium of the animals treated with GS10% was histopathologically analyzed, insertion periodontium was preserved. The results for these groups were significantly different of the vehicle group (p < 0.05). According to the results, the gel based in the aqueous extract of H. pectinata at 10% can prevent bone loss in experimental periodontal disease similarly to doxycycline 10%.

Evaluation of the toxicity and molluscicidal and larvicidal activities of Schinopsis brasiliensis stem bark extract and its fractions

Dengue fever and schistosomiasis are major public health issues for which vector control using larvicide and molluscicide substances present in plants provides a promising strategy. This study evaluated the potential toxicity of the extract of hydroethanol Schinopsis brasiliensis Engl., Anacardiaceae, stem bark and its chloroform, hexane, ethyl acetate, and hydromethanol fractions against Artemia salina and Aedes Aegypti larvae and snails Biomphalaria glabrata. All of the assays were performed in triplicate and the mean mortality rates were used to determine the LC50 and LC90 values using the probit method. The hydroethanol hydromethanol extract and fraction were free of toxicity towards A. salina (LC50 > 1000 µg/ml), while chloroform fraction was moderately toxic (LC50 313 µg/ml); ethyl acetate and hexane fractions displayed low toxicity, with LC50 557 and 582 µg/ml, respectively. Chloroform, hexane, and ethyl acetate fractions showed larvicidal potential towards A. aegypti (LC50 values of 345, 527 and 583 µg/ml, respectively), while chloroform and ethyl acetate fractions were highly toxic to B. glabrata (LC90 values of 68 and 73 µg/ml, respectively). Based on these findings, ethyl acetate, chloroform, and hexane fractions should be further investigated for their potential use against the vectors of dengue and schistosomiasis.

Orofacial antinociceptive effect and antioxidant properties of the hydroethanol extract of Hyptis fruticosa salmz ex Benth.

ETHNOPHARMACOLOGICAL RELEVANCE: Hyptis fruticosa is a plant native to Brazil with antinociceptive and antiinflamatory properties. This study evaluated the antinociceptive activity of the hydroethanol extract of the plant leaves (CHEE) against orofacial pain as well as its in vitro effect against lipid peroxidation. MATERIALS AND METHODS: The antinociceptive activity was investigated in mice orally treated with different doses of the CHEE (50, 100, and 200 mg/kg) and morphine (5 mg/kg) using formalin, glutamate, and capsaicin orofacial pain models using. Lipoperoxidation was induced in egg yolk by AAPH and FeSO4 in the absence and presence of the CHEE (5, 50, 100, and 150 µg/mL). RESULTS: CHEE (200 mg/kg) significantly reduced (ρ<0.001) the pain response in the first (69.6%) and second (81.8%) phases of the formalin test, while the nociception caused by capsaicin was significantly (ρ<0.001) reduced by up to 62% at 200 mg/kg of extract. When glutamate was used as algogen, a significant (ρ<0.001) nociception reduction of up to 85% at 200 mg/kg extract was observed. CHEE showed a higher protection against lipoperoxidation caused by FeSO4 (82.3% TBARS inhibition) than AAPH (35.7% TBARS inhibition) at 150 µg/mL. CONCLUSION: Hyptis fruticosa leaf CHEE is of pharmacological interest because it was able to inhibit the peripheral and central transmission of orofacial pain, while reducing the spreading of the inflammatory processes by neutralizing reactive oxygen species, which are by-products in the biosynthesis of pain mediators.

Atividade antimicrobiana de frações da própolis vermelha de Alagoas, Brasil / Antimicrobial activity of fractions of red propolis from Alagoas, Brazil

A própolis vermelha, como é conhecida popularmente, é uma própolis recentemente encontrada no Brasil e tem potente ação biológica. O presente trabalho avaliou a atividade antimicrobiana do extrato etanólico e das frações hexânica, clorofórmica e acetanólica da própolis proveniente de apiário do estado de Alagoas. As linhagens microbianas utilizadas foram: Shigella flexneri, Proteus vulgaris, Staphylococcus, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli e Candida albicans. O extrato etanólico apresentou atividade antimicrobiana frente a cepas gram-positivas (100%), gram-negativas (62,5%) e fúngicas (100%), com eficiência em 76,9% em todas as espécies testadas. A fração hexânica mostrou eficiência em 76,9% das espécies, semelhante ao extrato bruto; já a fração clorofórmica mostrou atividade frente a 92,3% das espécies analisadas, sendo Klebsiella pneumoniae a única espécie resistente. A fração acetanólica foi a fração que apresentou melhor atividade antimicrobiana com eficiência em 100% das espécies analisadas. Perante Candida albicans, observamos excelentes resultados, principalmente para a fração acetanólica, na qual a concentração inibitória mínima (CIM) se compara aos valores encontrados para as bactérias gram-positivas. Assim, as frações de própolis vermelha apresentaram excelente atividade antimicrobiana, principalmente frente a microrganismos gram-positivos e Candida albicans. Além disso, observamos que a fração acetanólica destacou-se como um promissor produto biotecnológico.

The red propolis is a new type of propolis founded in Brazil with large biological action. This study evaluated the antimicrobial activity of ethanol extract and fractions of hexane, chloroform and acetanolica of propolis from the apiary of the state of Alagoas. The microbial strains used were: Shigella flexneri, Proteus vulgaris, Staphylococcus, Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, Staphylococcus aureus, Escherichia coli and Candida albicans. The ethanolic extract showed activity against Gram-positive strains (100%), Gram-negative (62.5%) and fungi (100%) with efficiency against 76.9% of all the analyzed strains. The hexanic fraction showed efficiency against 76.9% as observed with the ethanolic extract, though the chloroform fraction showed activity against 92.3% of the strains analyzed. Klebsiella pneumoniae was the only resistant species. The ethyl acetate was the fraction that showed the best antimicrobial activity with efficiency against 100% of all strains analized. Excellent results were observed to Candida albicans mainly with ethyl acetate fraction with the value of minimum inhibitory concentration (MIC) similar to those observed to Gram-positives bacteria. We concluded that the partitions of red propolis showed excellent antimicrobial activity mainly against Gram-positive microorganisms and Candida albicans. Furthermore, we observed the ethyl acetate fraction stood out as a promissory biotechnological product.

Antinociceptive effects of an extract, fraction and an isolated compound of the stem bark of Maytenus rigida

The antinociceptive activity of the Maytenus rigida Mart. (Celastraceae) ethanol extract and its ethyl acetate fraction as well as of (-)-4'-methylepigallocatechin (1), a previously isolated compound, was demonstrated in vivo. ED50 for 1 in the writhing test was 14.14 mg/kg. The acetic acid-induced writhing was inhibited by 98.4, 84.4, and 58.3%, respectively, when mice were treated with the ethanol extract, ethyl acetate fraction, and 1. In the hot plate test, mice pretreated with 1 showed significantly increased reaction times (60-89%). Oral administration of 1 significantly inhibited first and second phases of the formalin-induced pain (50 and 26.5%, respectively), whereas indomethacin inhibited only the second phase of the test (41.2%). Ethanol extract and its fraction showed effects on inflammatory pain, while neurogenic and inflammatory pain suppression by 1 is a strong indication of the presence of both central and peripheral effects and suggests its analgesic and anti-inflammatory potential.

Anti-inflammatory and redox-protective activities of citronellal.

The anti-inflammatory and redox protective effects of the citronellal (CT) were evaluated using in vivo and in vitro tests. Intraperitoneal (i.p.) administration of CT (50, 100, and 200 mg/kg) inhibited (p < 0.05) the carrageenan-induced leukocyte migration to the peritoneal cavity. Additionally, the carrageenan- and arachidonic acid-induced rat hind paw edema was significantly inhibited (p < 0.05) by i.p. administration of 100 and 200 mg/kg of the compound. When the redox activity was evaluated, CT (200 mg/kg) significantly reduced hepatic lipoperoxidation (p < 0.001), as well as oxidation of plasmatic (p < 0.05) and hepatic (p < 0.01) proteins. The results of the present study support the hypothesis that CT possesses anti-inflammatory and redox protective activities. It is suggested that its effects are associated with the inhibition of the enzymes in the arachidonic acid pathway, which prevent cell migration by inhibiting leukotriene production, edema formation and the increase of reactive oxygen species in tissues. Therefore, CT is of potential benefit to manage inflammatory disorders and correlated damages caused by oxidant agents.

Chemical composition, acute toxicity, and antinociceptive activity of the essential oil of a plant breeding cultivar of basil (Ocimum basilicum L.).

Ocimum basilicum L. is an aromatic herb used in Brazil to treat illnesses such as respiratory and rheumatic problems, vomiting, and pain. In the present study, the chemical composition, acute toxicity, and antinociceptive effects of the essential oil (EO) of the cultivar "Maria Bonita" obtained from O. basilicum L. PI 197442 genotype were evaluated in Swiss mice (20-35 g each). Lethal dose to cause 50 % death (LD50) was calculated from a dose-response curve (100-5000 mg/kg body wt.; n = 6) as 532 mg/kg body wt. In the acetic acid-induced writhing test (0.6 % i. p.), EO (50, 100, and 200 mg/kg body wt., n = 8, s. c.) was effective in reducing the abdominal contractions at all doses (48-78 %). In the hot-plate test, EO significantly increased the latency at 50 mg/kg body wt. at all times (37-52 %, n = 8, s. c.). However, the effects of morphine and EO at 50 mg/kg were reverted in the presence of naloxone, an opioid antagonist. In the formalin test, EO significantly reduced paw licking time in the first and second phases of pain at 200 mg/kg body wt. (38 and 75 %, respectively, n = 8, s. c.). The results suggested that the peripheral and central antinociceptive effects of EO are related to the inhibition of the biosynthesis of pain mediators, such as prostaglandins and prostacyclins, and its ability to interact with opioid receptors.

Anti-inflammatory and redox-protective activities of citronellal

The anti-inflammatory and redox protective effects of the citronellal (CT) were evaluated using in vivo and in vitro tests. Intraperitoneal (i.p.) administration of CT (50, 100, and 200 mg/kg) inhibited (p < 0.05) the carrageenan-induced leukocyte migration to the peritoneal cavity. Additionally, the carrageenan- and arachidonic acid-induced rat hind paw edema was significantly inhibited (p < 0.05) by i.p. administration of 100 and 200 mg/kg of the compound. When the redox activity was evaluated, CT (200 mg/kg) significantly reduced hepatic lipoperoxidation (p < 0.001), as well as oxidation of plasmatic (p < 0.05) and hepatic (p < 0.01) proteins. The results of the present study support the hypothesis that CT possesses anti-inflammatory and redox protective activities. It is suggested that its effects are associated with the inhibition of the enzymes in the arachidonic acid pathway, which prevent cell migration by inhibiting leukotriene production, edema formation and the increase of reactive oxygen species in tissues. Therefore, CT is of potential benefit to manage inflammatory disorders and correlated damages caused by oxidant agents.

Perfil fitoquímico e ensaio microbiológico dos extratos da entrecasca de Maytenus rigida Mart. (Celastraceae) / Phytochemistry and microbiological assay of the bark extracts of Maytenus rigida Mart. (Celastraceae)

Maytenus rigida Mart (Celastraceae), conhecida por "Bom-homem", "Bom-nome", "Cabelo-de-negro", "Casca-grossa" e "Pau-de-colher", é uma arvore de pequeno porte. A entrecasca do caule é empregada popularmente no Nordeste do Brasil no tratamento das dores em geral, infecções e inflamações. O presente trabalho avaliou tanto o perfil fitoquímico de M. rigida por meio de um roteiro analítico, quanto à atividade antibiótica dos extratos pelo método de Kirby-Bauer modificado. Os resultados demonstraram que os extratos etanólico, aquoso, clorofórmico, acetato de etila e hidroalcoólico de M. rigida apresentam atividade antibacteriana contra Escherichia coli, Pseudomonas aeruginosa e Staphylococcus aureus, enquanto que a fração hexânica não exibe qualquer atividade. Catequinas, quinonas, esteróides, triterpenos, saponinas, flavonóides e compostos fenólicos foram detectados na análise fitoquímica.

Maynetus rigida Mart (Celastraceae), known as "Bom-homem", "Bom-nome", "Cabelo-de-negro", "Casca-grossa" and "Pau-de-colher", is a small tree. The stem bark is used by the population in the northeast of Brazil to treat aches, infections and inflammations in general. The present work evaluated both the phytochemistry of M. rigida Mart by an analytical routine, and the antimicrobial activity of the bark extracts by the Kirby-Bauer modified method. Our results showed the aqueous, methanol, chloroform, ethyl acetate and hydroalcoolic extracts of M. rigida Mart has antimicrobial activity against Escherichia coli, Pseudomonas aeruginosa and Staphylococcus aureus, while the hexane extract does not have any activity. Catechins, quinones, steroids, triterpenes, saponins, flavonoids and phenolic compounds were detected by the phytochemical analysis.

Uptake and transformation of phenol and chlorophenols by hairy root cultures of Daucus carota, Ipomoea batatas and Solanum aviculare.

Hairy root cultures of Daucus carota L., Ipomoea batatas L. and Solanum aviculare Forst were investigated for their susceptibility to the highly toxic pollutants phenol and chlorophenols and for the involvement of inherent peroxidases in the removal of phenols from liquid media. Roots of D. carota grew normally in medium containing 1000 micromol l(-1) of phenol, whilst normal growth of roots of I. batatas and S. aviculare was only possible at levels up to 500 micromol l(-1). In the presence of chlorophenols, normal root growth was possible only in concentrations not exceeding 50 micromol l(-1), except for I. batatas which was severely affected at all concentrations. Despite the reduction in biomass, the growth of S. aviculare cultures was sustained in medium containing up to 2000 micromol l(-1) of phenol or 2-chlorophenol, and up to 500 micromol l(-1) of 2,6-dichlorophenol. The amounts of phenol removed by the roots within 72 h of treatment were 72.7%, 90.7% and 98.6% of the initial concentration for D. carota, I. batatas and S. aviculare, respectively. For the removal of 2,6-dichlorophenol the values were, respectively, 83.0%, 57.7% and 73.1%. Phenols labelled with 14C were absorbed by the root tissues and condensed with highly polar cellular substances as well as being incorporated into the cell walls or membranes. The results suggest that S. aviculare, an ornamental plant, would be best suited for remediation trials under field conditions.

Tolerance and metabolism of phenol and chloroderivatives by hairy root cultures of Daucus carota L.

Hairy root cultures are shown to be suitable experimental systems to screen higher plants for tolerance to various inorganic and organic pollutants, and for determining the role of the root matrix in the uptake and further metabolism of contaminants. A number of clones were obtained by infection of carrot tissues with Agrobacterium rhizogenes and two (the fastest and the slowest growing root clones) were chosen for further experimentation. Both clones showed a similar degree of tolerance towards phenol and its chlorinated derivatives, i.e. the growth of root biomass was maintained in concentrations of phenol equivalent to 1000 micromol/l, whilst the chlorophenols were tolerated only at concentrations 20 times lower (50 micromol/l). Transformed carrot roots were able to remove more than 90% of the exogenous phenolic compounds from the culture medium within 120 h after treatment. Metabolism of these compounds occurred in the root tissue and was accompanied by an increase in peroxidase activity.