Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark / Caracterização fitoquímica, atividade antioxidante e antibacteriana do extrato hidroetanólico de cascas do caule de Anadenanthera peregrina

The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-¹. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.

O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físico química, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência(HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.

Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark

Abstract The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-1. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.

Resumo O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físico-química, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência (HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.

Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark / Caracterização fitoquímica, atividade antioxidante e antibacteriana do extrato hidroetanólico de cascas do caule de Anadenanthera peregrina

The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-1. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.

O bioma Cerrado brasileiro apresenta em uma grande variedade de espécies endêmicas com diversos compostos bioativos, e Anadenanthera peregrina (L.) Speg é uma espécie promissora. Neste estudo, objetivamos realizar a caracterização fitoquímica e avaliar as atividades antioxidantes e antibacterianas contra Staphylococcus aureus e Escherichia coli do extrato hidroetanólico de cascas do caule de A. peregrina. As cascas foram coletadas no Jardim Botânico de Goiânia, Brasil. O extrato hidroetanólico foi obtido por percolação e submetido a triagem físicoquímica, estimativa de conteúdo fenólico total, impressão digital por cromatografia líquida de alta eficiência (HPLC) e determinação da atividade antioxidante (valores de IC50 foram calculados para o ensaio 2,2-difenil-1-picril-hidrazil) e antibacteriana. O pH do extrato foi de 5,21 e a densidade foi de 0,956 g/cm3. A triagem fitoquímica indicou a presença de glicosídeos cardíacos, ácidos orgânicos, açúcares redutores, saponinas hemolíticas, fenóis, cumarinas, taninos condensados, flavonóides, catequinas, depsídios e depsidonas derivados de benzoquinonas. O extrato mostrou intensa atividade hemolítica. O conteúdo fenólico total foi de 6,40 g de GAE 100 g-1. A análise por impressão digital por HPLC revelou a presença de ácido gálico, catequina e epicatequina. Confirmamos a atividade antioxidante do extrato. Além disso, o extrato não inibiu o crescimento de colônias de E. coli em nenhum volume testado, mas houve halos em torno das colônias de S. aureus nos três volumes testados. Estes resultados contribuem para uma melhor compreensão da composição química da casca de A. peregrina e apoia ainda mais as aplicações medicinais desta espécie.

Phytochemical characterization, and antioxidant and antibacterial activities of the hydroethanolic extract of Anadenanthera peregrina stem bark.

The Brazilian Cerrado biome consists of a great variety of endemic species with several bioactive compounds, and Anadenanthera peregrina (L.) Speg is a promising species. In this study, we aimed to perform phytochemical characterization and evaluate the antioxidant and antibacterial activities against Staphylococcus aureus and Escherichia coli of the hydroethanolic extract of A. peregrina stem bark. The barks were collected in the Botanical Garden of Goiânia, Brazil. The hydroethanolic extract was obtained by percolation and subjected to physicochemical screening, total phenolic content estimation, high-performance liquid chromatography (HPLC) fingerprinting, and antioxidant (IC50 values were calculated for the 2,2-diphenyl-1-picrylhydrazyl assay - DPPH) and antibacterial activity determination. The pH of the extract was 5.21 and density was 0.956 g/cm3. The phytochemical screening indicated the presence of cardiac glycosides, organic acids, reducing sugars, hemolytic saponins, phenols, coumarins, condensed tannins, flavonoids, catechins, depsides, and depsidones derived from benzoquinones. The extract showed intense hemolytic activity. The total phenolic content was 6.40 g GAE 100 g-1. The HPLC fingerprinting analysis revealed the presence of gallic acid, catechin, and epicatechin. We confirmed the antioxidant activity of the extract. Furthermore, the extract did not inhibit the growth of E. coli colonies at any volume tested, but there were halos around S. aureus colonies at all three volumes tested. These results contribute to a better understanding of the chemical composition of A. peregrina stem bark and further support the medicinal applications of this species.

Low-level laser therapy improves peri-implant bone formation: resonance frequency, electron microscopy, and stereology findings in a rabbit model.

Previous studies have reported positive effects of low-level laser therapy (LLLT) on bone healing. This study evaluated the effects of LLLT on peri-implant healing in vivo. Thirty-two rabbits had their mandibular left incisors removed, followed by immediate insertion of a dental implant into the fresh socket. Animals were assigned randomly to four groups: control (non-irradiated) or LLLT at three different doses per session: 5J/cm(2), 10J/cm(2), and 20J/cm(2). A GaAlAs laser (830nm, 50mW) was applied every 48h for 13 days, starting immediately after surgery. The implant stability quotient (ISQ) was measured using resonance frequency analysis upon implant insertion and immediately after death, 30 days after the last application. Tissues were prepared for scanning electron microscopy (SEM) and stereology. Variables measured were bone-implant contact (BIC) and bone neoformation within implant threads at three different sites. The results showed better ISQ for the 20J/cm(2) group (P=0.003). BIC values were significantly higher (P<0.05) in the 20J/cm(2) group, on both SEM and stereology. Bone area values were better in the 10J/cm(2) (P=0.036) and 20J/cm(2) (P=0.016) groups compared to the control group. Under these conditions, LLLT enhanced peri-implant bone repair, improving stability, BIC, and bone neoformation. The findings support and suggest parameters for the design of clinical trials using LLLT after implant placement.

Evaluation of anti-nociceptive and anti-inflammatory activity of low-level laser therapy on temporomandibular joint inflammation in rodents.

The aim of this study was to investigate the analgesic and anti-inflammatory activity of low-level laser therapy (LLLT) on the nociceptive behavioral as well as histomorphological aspects induced by injection of formalin and carrageenan into the rat temporomandibular joint. The 2.5% formalin injection (FRG group) induced behavioral responses characterized by rubbing the orofacial region and flinching the head quickly, which were quantified for 45 min. The pretreatment with systemic administration of diclofenac sodium-DFN group (10 mg/kg i.p.) as well as the irradiation with LLLT infrared (LST group, 780 nm, 70 mW, 30 s, 2.1 J, 52.5 J/cm(2), GaAlAs) significantly reduced the formalin-induced nociceptive responses. The 1% carrageenan injection (CRG group) induced inflammatory responses over the time-course of the study (24 h, and 3 and 7 days) characterized by the presence of intense inflammatory infiltrate rich in neutrophils, scanty areas of liquefactive necrosis and intense interstitial edema, extensive hemorrhagic areas, and enlargement of the joint space on the region. The DFN and LST groups showed an intensity of inflammatory response that was significantly lower than in CRG group over the time-course of the study, especially in the LST group, which showed exuberant granulation tissue with intense vascularization, and deposition of newly formed collagen fibers (3 and 7 days). It was concluded that the LLLT presented an anti-nociceptive and anti-inflammatory response on the inflammation induced in the temporomandibular joint of rodents.

Assessment of the systemic effects of low-level laser therapy (LLLT) on thyroid hormone function in a rabbit model.

The aim of this study was to assess the effects of low-level laser therapy (LLLT) applied to a dental extraction socket on thyroid gland function in a rabbit model, based on serum triiodothyronine and thyroxine levels. Sixteen male New Zealand rabbits were randomly distributed into two groups: a control group (non-irradiated animals) and an experimental group (irradiated animals: one irradiation point in the extraction socket of the lower incisor). Animals in the experimental group were irradiated with an aluminium gallium arsenide diode laser (AlGaAs; wavelength 830 nm, 40 mW, CW laser), for 13 days, every 48 h, at a dose of 6 J/cm(2) per session, resulting in a total dose of 42 J/cm(2). Serum triiodothyronine and thyroxine levels were measured in both groups before extraction and on the last day of observation (day 15). There were no statistically significant differences between the groups in pre- and post-irradiation triiodothyronine and thyroxine values. With the irradiation protocol used in this study, LLLT did not affect thyroid function in rabbits as assessed by circulating serum triiodothyronine and thyroxine levels.

Hyptis pectinata: redox protection and orofacial antinociception.

Hyptis pectinata L. Poit, known as 'sambacaitá', is used in Brazil to treat inflammatory and painful disorders. In this study, the antioxidant and orofacial antinociceptive properties of the aqueous extract of H. pectinata leaves (AEPH) were assessed using in vitro and in vivo models. Thus, AEPH reduced the 2,2-diphenyl-1-picrylhydrazyl radical up to 72.10% with an EC50 of 14.56 µg/ml. It also inhibited 40.80% of the lipoperoxidation induced by 2'-azobis (2-amidinopropane) dihydrochloride in the thiobarbituric acid-reactive substances assay. The orofacial antinociceptive activity was evaluated in mice pre-treated with AEPH (100, 200 and 400 mg/kg, p.o.) and morphine (5 mg/kg, i.p.), which received afterwards formalin- (20 µl, 2% solution, s.c.), glutamate- (40 µl, 25 mM, s.c.) and capsaicin- (20 µl, 2.5 µg, s.c.) to induce orofacial nociception. AEPH at all doses reduced (p < 0.001) the nociceptive response in the first (43-62%) and second (47-80%) phases of the formalin test. Besides, the effect of AEPH (400 mg/kg) was not changed in the presence of naloxone (1.5 mg/kg, i.p.), an opioid antagonist. AEPH significantly inhibited mice face rubbing for capsaicin (23-69%, p < 0.05) and glutamate (48-77%, p < 0.001) at all doses. The findings suggested the AEPH has peripheral and central antinociceptive activities, which are not related to opioid receptors.

Sida cordifolia leaf extract reduces the orofacial nociceptive response in mice.

In this study, we describe the antinociceptive activity of the ethanol extract (EE), chloroform (CF) and methanol (MF) fractions obtained from Sida cordifolia, popularly known in Brazil as "malva branca" or "malva branca sedosa". Leaves of S. cordifolia were used to produce the crude ethanol extract and after CF and MF. Experiments were conducted on Swiss mice using the glutamate and formalin-induced orofacial nociception. In the formalin test, all doses of EE, CF and MF significantly reduced the orofacial nociception in the first (p < 0.001) and second phase (p < 0.001), which was also naloxone-sensitive. In the glutamate-induced nociception test, only CF and MF significantly reduced the orofacial nociceptive behavior with inhibition percentage values of 48.1% (100 mg/kg, CF), 56.1% (200 mg/kg, CF), 66.4% (400 mg/kg, CF), 48.2 (200 mg/kg, MF) and 60.1 (400 mg/kg, MF). Furthermore, treatment of the animals with EE, CF and MF was not able to promote motor activity changes. These data demonstrate that S. cordifolia has a pronounced antinociceptive activity on orofacial nociception. However, pharmacological and chemical studies are necessary in order to characterize the responsible mechanisms for this antinociceptive action and also to identify other bioactive compounds present in S. cordifolia.

Heteropterys aphrodisiaca (extract BST0298): a Brazilian plant that improves memory in aged rats.

Literature report is lacking on pharmacological studies of the plant Heteropterys aphrodisiaca, endemic to the scrublands of Brazil. The present study was carried out to investigate the effects of oral dosing with extract BST0298 from this plant, on learning and on memory, in young (3-6-month-old) and aged (20-28-month-old) rats. The aged animals presented significant memory deficits in both the passive avoidance and T-maze left/right discrimination tests. Treatment for 7 days (50 mg/kg) or 26 days (100 mg/kg) with extract BST0298 restored the memory deficits in the passive avoidance test. However, no improvement in memory was observed after acute administration of extract BST0298 (100 mg/kg) in aged rats. An improvement in learning was also observed in the left/right discrimination test in aged rats treated for 109 days with BST0298 at a dose of 50 mg/kg. These results suggest that treatment for 7 days or more with H. aphrodisiaca improves learning and memory deficits in aged rats.

Effect of vitamin E supplementation on antibody levels against malondialdehyde modified LDL in hyperlipidemic hamsters.

OBJECTIVE: The aim of this work was to investigate the effect of vitamin E (VE) supplementation on the formation of autoantibodies against oxidized low density lipoproteins (LDL) in a hyperlipidemic animal model. METHODS: Thirty-four male hamsters (Mesocricetus auratus), (4 weeks old) were divided into three groups: Group A (n=9) was fed with standard rodent chow; group B (n=13) was fed with a standard rodent chow plus 2% cholesterol and 10% butter and group C (n=12) was fed with the same diet plus 0.2% (w/w) VE. Blood samples were collected by intracardiac puncture and antibody levels were determined in each animal at 4 weeks of age and after 20 weeks of experimental diet. A modified ELISA technique was used to analyze the modulation of autoantibody titers against an epitope of oxidized LDL in serum samples. Antigens prepared for the ELISA tests were characterized using spectrofluorimetry. Serum VE levels were determined in the lipidic fractions by HPLC. RESULTS: The groups fed with cholesterol-fat enriched diet presented a three-fold increase in total serum cholesterol and two-fold increase in serum triglycerides compared to the control group. VE supplementation played no role in serum cholesterol and serum triglyceride concentrations but led to a decreased autoantibody (anti-LDL-malondialdehyde) formation (P<0.05). CONCLUSIONS: Our results show that VE supplementation leads to a lower production of autoantibodies against oxidized LDL, suggesting a protective effect of VE against in vivo oxidation of LDL particles, in a dose-dependent manner.

Antiulcerogenic effects of two Maytenus species in laboratory animals.

Leaves of Maytenus species are commonly used in Brazil for the treatment of gastric ulcers, dyspepsias and other gastric problems. The present study evaluated the antiulcerogenic potential of a boiling water extract of equal parts of M. aquifolium and M. ilicifolia leaves against ulcer lesions induced by indomethacin and cold-restraint stress in rats. Ranitidine and cimetidine were used as reference drugs. The oral and intraperitoneal administration of the extract had a potent antiulcerogenic effect against both types of ulcers. The extract was shown to cause an increase in volume and pH of gastric juice of the animals with the pH effects comparable to those of cimetidine. The results tend to confirm the popular use of the plant.

Pharmacologic and toxicologic effects of two Maytenus species in laboratory animals.

Leaves of Maytenus species are used in the popular medicine of Brazil for their reported antacid and antiulcerogenic activity. The present work examined the effects of a boiling water extract of equal parts of M. aquifolium and M. ilicifolia leaves on acute administration in rats and mice, in an attempt to detect any general depressant, hypnotic, anticonvulsant and analgesic effects. General depressant and hypnotic effects were seen only after intraperitoneal administration. After chronic administration, the overall behavior of animals did not change and they continued to gain weight at the same rate as controls. Several biochemical and hematological parameters as well as pathological examination of different organs did not show any significant alterations after 3 months of treatment. A search for the potential effects of the extract on the fertility of female and male rats and on the course of pregnancy as well as a search for potential teratogenic effects did not reveal any significant differences from controls. Taken together, the results indicate that these Maytenus species may be safe for human use and deserve further investigation.