Geographical variation in the chemical composition, anti-inflammatory activity of the essential oil, micromorphology and histochemistry of Schinus terebinthifolia Raddi.

ETHNOPHARMACOLOGICAL RELEVANCE: Schinus terebinthifolia Raddi (Anacardiaceae) is rich in essential oil, distinguished by a predominance of monoterpenes and sesquiterpenes, it being widely used in traditional medicine for the treatment of inflammations. AIM OF STUDY: This study's objective was to investigate the chemical composition of the essential oil of S. terebinthifolia (EOST) collected in six states of Brazil, evaluate its anti-inflammatory effects in mice, and analyze the histochemistry and micromorphology of leaves and stems. MATERIALS AND METHODS: Aerial parts of S. terebinthifolia were collected in six states of Brazil, and the essential oil was extracted by hydrodistillation and analyzed by gas chromatography-mass spectrometry (GC-MS). The histochemistry and micromorphology of leaves and stems were performed using standard reagents, light and field emission scanning electron microscopy, beyond energy-dispersive X-ray spectroscopy. The EOST were evaluated for anti-inflammatory activity and hyperalgesia using the carrageenan-induced paw edema methodology. RESULTS: The EOST showed variation across the six states in its yield (0.40%-0.86%) and chemical composition: hydrocarbon monoterpenes (28.76%-47.73%), sesquiterpenes, (31.43%-41.76%), oxygenated monoterpenes (14.31%-19.57%), and oxygenated sesquiterpenes (4.87%-14.38%). Both α-pinene and limonene were predominant constituents of essential in five regions, except for one state where α-phellandrene and limonene were the dominant components. A comprehensive description of the leaf and stem micromorphology and histochemistry was performed. In the in vivo testing, all EOST samples exerted antiedematogenic and anti-hyperalgesic effects, when tested in a carrageenan-induced paw inflammation (mechanical and thermal hyperalgesia) model with oral doses of 30 mg/kg. CONCLUSION: Our results indicate that the EOST samples collected in six Brazilian states differed in their chemical composition but not their anti-inflammatory and antihyperalgesic effects, which was correlated with the synergistic effect of its components, collaborating the etnhopharmacologycal use of this plant due to its an anti-inflammatory effect. Also, micromorphology and histochemistry of leaves and stems presented in this study provide anatomical and microchemical information, which aids species identification.

Exploration of essential oil from Psychotria poeppigiana as an anti-hyperalgesic and anti-acetylcholinesterase agent: Chemical composition, biological activity and molecular docking.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves from Psychotria poeppigiana Müll. Arg. (accepted as Palicourea tomentosa (Aubl.) Borhidi), Rubiaceae, has traditionally been used in medicine for treatments of inflammation and pain; Synonymously, Cephaelis elata for the treatment of dementia; However, few scientific studies have been evidence demonstrating this activity. AIM OF THE STUDY: The aim of this study was to investigate the chemical composition of P. poeppigiana essential oil obtained from leaves (EOPP) and its antioxidant, anti-inflammatory and acetylcholinesterase (AChE) activities. Molecular docking simulations were carried out with the main constituents. MATERIALS AND METHODS: EOPP (hydrodistillation) was analysed by gas chromatography-mass spectrometry (GC-MS). The fractionation of EOPP afforded germacrene D and bicyclogermacrene. The antioxidant activity of EOPP was determined by MDA assay. The inflammatory parameters were evaluated using CFA model (with paw edema, mechanical, thermal hyperalgesia, MPO and NAG) in EOPP (30, 100 and 300 mg/kg), germacrene D and bicyclogermacrene (30 mg/kg). The AChE inhibition was evaluated in rat brain structures and molecular docking simulations were carried out using Autodock v.4.3.2. RESULTS: GC-MS analysis identified 19 compounds, and the major compounds were germacrene D (29.38%) and bicyclogermacrene (25.21%). EOPP exhibited high antioxidant capacity (IC50 = 12.78 ± 1.36 µg/mL). All the tested doses of EOPP and both major constituents significantly inhibited cold and mechanical hyperalgesia and significantly blocked the increase in MPO activity 24 h after the CFA injection. There was significant AChE inhibition by EOPP and germacrene D in the cerebral cortex and hippocampus (>50%). Enzyme-ligand molecular modelling showed that the major constituents of EOPP interacted differently with AChE. CONCLUSIONS: The chemical compounds of the essential oil from the leaves of P. poeppigiana is based mainly on terpenes, the sesquiterpenes germacrene D (29.38%) and bicyclogermacrene (25.21%) being the major compounds. EOPP presented antioxidant, anti-inflammatory and anti-acetylcholinesterase (AChE) activities. Besides, enzyme-ligand molecular modelling showed the EOPP may act as an anti-hyperalgesic and AChE inhibitory agent. Taken together, these results might be in accordance with if folk use for pain- and inflammation-related symptoms.

Contribution of spathulenol to the anti-nociceptive effects of Psidium guineense.

Objectives: Araçá-verdadeiro is the popular name of Psidium guineense (Myrtaceae), whose fruits and leaves are used in Brazilian folk medicine for treatment of inflammation and pain. The focus of the present research was an investigation of the anti-nociceptive, and anti-inflammatory effects of the essential oil from P. guineense (EOPG) leaves, and of spathulenol. The anxiolytic and antidepressive effects associated with chronic pain were also investigated in models of acute or persistent nociception or/and inflammatory pain.Methods and Results: Oral treatment with EOPG (10-100 mg/kg) or spathulenol (10 mg/kg) significantly inhibited formalin-induced nociceptive responses, both sensitivity to cold and edema. Oral treatment with EOPG (10 mg/kg) and spathulenol (10 mg/kg) did not reduce locomotor activity (open field test). Local administration of spathulenol (1000 µg/paw) significantly prevented formalin-induced nociceptive sensitivity to cold and paw edema, and carrageenan-induced mechanical hyperalgesia, paw edema and sensitivity to cold. In the Freund's complete adjuvant (CFA) model, oral treatment with EOPG (10 mg/kg) or spathulenol (10 mg/kg) for 21 days significantly inhibited all analyzed parameters. The percentage maximal inhibition by spathulenol was 76.00% (mechanical hyperalgesia), 71.90% (cold response), 85.00% (edema), 77.16% (myeloperoxidase activity), 97.72% (time in the closed arms in the elevated plus maze), and 49.00% (immobility time in the tail suspension test), in the CFA model. Models employed male Swiss mice, except for the CFA test, which employed C57bL6 male mice (n=6 /group).Conclusion: This study demonstrates that EOPG is an anti-nociceptive and anti-hyperalgesic agent, in acute and continuous treatment, and an anxiolytic and antidepressive agent when tested with the chronic pain experimental state.

Variation in essential oil components and anti-inflammatory activity of Allophylus edulis leaves collected in central-western Brazil.

ETHNOPHARMACOLOGICAL RELEVANCE: An infusion obtained from the leaves of "chal-chal" (Allophylus edulis Radlk.) is used for popular treatment of intestinal disorders and as an anti-inflammatory throat treatment. Because of the anti-inflammatory medicinal folk use, a previous work reported scientific research confirming the anti-inflammatory activity of A. edulis essential oil collected in Dourados, MS, Brazil, in March 2015. AIM OF THE STUDY: The aim of this study was to evaluate the variation in the chemical profile of the essential oil of A. edulis plants collected in Dourados (EOAE-D) and Bonito (EOAE-B), two cities in Mato Grosso do Sul State, Brazil. Additionally, we evaluated the anti-inflammatory effects of the essential oil, as well as that of the major compounds (caryophyllene oxide and α-zingiberene), in experimental in vivo models of inflammation in mice. MATERIALS AND METHODS: Leaves were collected from plants at both sites in July 2018. The composition of the essential oil (EOAE-D and EOAE-B) was determined by GC/MS, and major compounds (caryophyllene oxide and α-zingiberene) were isolated and identified by chromatographic methods and NMR spectroscopy. Anti-inflammatory capacities were assessed using two classical models of inflammatory models, carrageenan- and CFA-induced paw inflammation (mechanical and thermal hyperalgesia). RESULTS: Both EOAE-D and EOAE-B showed sesquiterpenes as a major constituent, namely, caryophyllene oxide (29.5%) and α-zingiberene (45.0%), respectively. In tests, EOAE, caryophyllene oxide and α-zingiberene-induced antiedematogenic and antihyperalgesic effects were found in the different utilized models. CONCLUSIONS: The results indicate that samples from the two cities differed in chemical composition but not in their anti-inflammatory and antihyperalgesic effects. This finding corroborates the use of A. edulis as a medicinal plant and indicates its potential in the therapy of inflammatory conditions.

Antimicrobial activity of plant extracts against carbapenem-producing Klebsiella pneumoniae and in vivo toxicological assessment.

The global spread of multidrug-resistant strains has prompted the scientific community to explore novel sources of chemicals with antimicrobial activity. The aim of the study was to examine the antimicrobial activity in vitro of 28 extracts against carbapenem-producing Klebsiella pneumoniae, individually and in combination with antibiotics and in vivo toxicological assessment of the most active product. The multi-resistant K. pneumoniae strain was submitted for phenotypic and molecular characterization. The antibacterial activity of 28 plant extracts was evaluated alone and in combination with antibiotics against this strain through the agar disk diffusion. Of these, 16 extracts showed synergism against carbapenem-producing K. pneumoniae, being that B. crassifolia extract exhibited synergism with three antibiotics. Based on this assessment, B. crassifolia-extract-induced toxicity on Swiss male mice was evaluated by administering this extract and subsequently determining apoptosis and splenic phagocytosis using the comet and micronucleus assays. The results of this study showed that B. crassifolia extract had synergistic activity promising and groups treated with B. crassifolia exhibited no genotoxic or mutagenic activity, indicating that B. crassifolia extract exerted beneficial effects and appeared safe to use at the studied concentrations.

Effects of Bidens gardneri Baker leaves aqueous extract in glucose homeostasis of rats.

Bidens gardneri Baker, popularly known as "picão-vermelho", has been used, traditionally, as a medicinal plant for the treatment of Diabetes mellitus. This study aimed to evaluate antidiabetic effect of leaves from B. gardneri aqueous extract (BGAE). We also evaluated in vitro anti-glycation potential. Chemical composition was analyzed based on a colorimetrics and HPLC methods. The oral glucose tolerance test (OGTT), was performed in rats with different doses (30, 100 and 300 mg/kg). Alloxan-induced diabetic and hypercaloric diet-fed rats were treated with 300 mg/kg of BGAE, orally, for 10 days and then 10 weeks, respectively. The activity of intestinal disaccharidases (maltase, sucrase and lactase) and quantification of muscle and liver glycogen content were also evaluated. On chemical analysis, the extract showed high phenolics content and the chromatogram showed 4-O-caffeoylquinic acid as the major component. The extract presented inhibition in the formation of advanced glycation end products (AGEs) and disaccharidases activity. In OGTT the dose of 300 mg/kg significantly decreased the blood glucose. In diabetic animals, BGAE significantly decreased blood glucose levels, preventing weight loss. In addition, in hypercaloric diet-fed rats, the extract prevented hyperglycemia. Our results suggest that, aqueous extract of B. gardneri has a potential for therapeutic intervention of diabetes.

Bioguided Fractionation, and Antioxidant, Antiproliferative, and Anti-Inflammatory Activity of Annona cacans Warm.

This study evaluated to determine the phenolic and flavonoids contents, and antioxidant, anti-inflammatory, and antiproliferative activity of the hydromethanolic extracts of the leaves, pulp, and seeds of Annona cacans. The isolation and structural identification of the constituent acetogenin, phenolic acid, and flavonoids were also reported. Antioxidant capacity was determined by the 2,2-diphenyl-1-picrylhydrazyl (DPPH), ethylbenzothiazoline-6-sulfonic acid (ABTS), and ß-carotene/linoleic acid methods. Cell proliferation was determined by spectrophotometric quantification of the cellular protein content using a sulforhodamine B assay. Anti-inflammatory activity was evaluated in paw edema model, to myeloperoxidase (MPO) activity induced by carrageenan in mice. Fractionation resulted in the isolation of one acetogenin (annoreticuin-9-one), two flavonoids (quercetin-3-O-ß-glucoside-6-O-α-rhamnoside and kaempferol-3-O-ß-glucoside), and one phenolic acid (p-coumaric acid). The pulp extract presented potent antioxidant activities by the DPPH (IC50 = 44.08 µg/mL) and ABTS (IC50 = 39.32 µg/mL) methods, as well as high contents of phenols (618.95 mg GA/g) and flavonoids (477.35 mg QE/g). The bioguided fractionation demonstrated that the ethyl acetate fraction of the pulp extract and annoreticuin-9-one showed potent antiproliferative activity against ovarian cancer (GI50 = 6.4 µg/mL). The anti-inflammatory activity demonstrated significant inhibition of edema compared to the control group in 2 and 4 h; in addition, the extracts inhibited the increase in MPO activity after 6 h, when compared to the DEX and control groups. For the first time, this study demonstrated antioxidant, anti-inflammatory, and antiproliferative activity, as well as compounds isolated, suggesting that A. cacans could also be potential sources for prevention of cancer and other diseases associated with oxidative stress.

Psychotria leiocarpa Extract and Vincosamide Reduce Chemically-Induced Inflammation in Mice and Inhibit the Acetylcholinesterase Activity.

Species from Psychotria are used in folk medicine against inflammatory diseases, respiratory disturbances, and anti-hallucinogenic. In the present study, the compound vincosamide (PL-1) was identified for the first time in methanolic extract of the Psychotria leiocarpa (ME-PL) leaves, as well as the anti-inflammatory and anticholinesteric effects in rodents and molecular docking simulations. The fractionation of the chloroform fraction (CF-PL) through chromatographic methods afforded the known compound PL-1. The anti-inflammatory activity of the ME-PL (30, 100, and 300 mg/kg) and PL-1 (3, 30, and 100 mg/kg) was analyzed using experimental models: paw edema, pleurisy, and mechanical and thermal hyperalgesia induced by carrageenan. The anticholinesterase activity of the ME-PL (30 and 100 mg/kg) and PL-1 (30 mg/kg) was showed by acetylcholinesterase (AChE) inhibitory in brain structures. The molecular docking simulations were performed using Molegro Virtual Docker v6.0. Overall, the results indicated that ME-PL and PL-1 demonstrated an anti-edematogenic effect in Cg-induced paw edema, leukocyte migration in the pleurisy model, and significantly reduced mechanical hyperalgesia, cold response to acetone in mice. The samples exhibited maximal inhibition of enzyme acetylcholinesterase (AChE) in the frontal cortex. The molecular coupling of PL-1 with the AChE showed significant interactions with the catalytic and peripheral site, corroborating the activity presented in the inhibition assay. The acute administration of ME-PL did not cause signs of toxicity in the treated animals. The results showed that P. leiocarpa inhibited AChE and anti-inflammatory activity, and alkaloid vincosamide could be responsible, at least in part, for the observed effects, supporting the popular use of this genus.

Anti-inflammatory Activity of Methanolic Extract and an Alkaloid from Palicourea crocea (Sw.) Roem and Schult.

Palicourea crocea (Sw.) Roem. and Schult., "douradinha," are used by treat inflammation (edema). Croceaine A (PC-1) was isolated from P. crocea (MEPC) leaves and studied for its antioxidant and anti-inflammatory activity, as well as concentrations of constituents and acute toxicity. The phenols and polyphenolics compounds and HPLC/DAD were determined. The antioxidant activity were evaluated for DPPH, ABTS, and MDA. MEPC (300, 100, and 300 mg/kg) and PC-1 (10 and 30 mg/kg) were tested for anti-inflammatory effects in paw edema, pleurisy, cold sensitivity, and mechanical hyperalgesia. Acute toxicity is also described. MEPC contained high concentrations of phenolic and flavonoid compounds (≤ 800.35 mg/g), as well as caffeic acid, ferulic acid, rutin, and quercetin, revealed by HPLC-DAD analysis. MEPC displayed antioxidant activity against ABTS radicals (IC50 = 68.5 µg/mL) and MDA (74%). MEPC and alkaloid PC-1 demonstrated an anti-edematogenic effect in Cg-induced paw edema in 2 and 4 h, and also significantly reduced mechanical hyperalgesia, cold response to acetone in mice, at 3 and 4 h after injection, as well as leukocyte migrationin the pleurisy model. No toxicity was detected by MEPC. For the first time, P. crocea was evaluated for its antioxidant, systemic anti-inflammatory, and anti-hyperalgesic activities.

Chemical constituents of Cochlospermum regium (Schrank) Pilg. root and its antioxidant, antidiabetic, antiglycation, and anticholinesterase effects in Wistar rats.

The main physiological consequence of diabetes mellitus is chronic hyperglycemia. This condition is related to the formation of free radicals including advanced glycation end products (AGES) and to an increase in inflammatory processes. Cochlospermum regium (Schrank) Pilg., part of the Bixaceae family, is a cerrado plant known for its anti-inflammatory effects. The objectives of this study were to analyze the constituent compounds of C. regium roots and to evaluate the antioxidant, antiglycation, antidiabetic, and anticholinergic effects of its hydromethanolic extract through in vitro and in vivo experimental models. The presence of phenols, flavonoids, condensed tannins, and flavonols was analyzed by liquid chromatography - photodiode array (LC/PDA) analysis. Whereas antioxidant activity was investigated via DPPH, ABTS, ß-carotene/linoleic acid, and malondialdehyde colorimetric assays. Inhibition of AGEs was examined via a bovine serum albumin system whose glycosylating agent was glucose. Antidiabetic potential was examined in normoglycemic Wistar rats that received glucose overload, in alloxan-induced diabetic rats, and in rats that received a hyperglycemic diet. Disaccharidase inhibition was assessed using in vitro and in vivo methods, as was acetylcholinesterase (AChE) inhibition in brain structures. The hydromethanolic extract (CRHE) possessed a high concentration of phenolic compounds and showed antioxidant effects. The LC-DAD results revealed that CRHE contained a high concentration of phenolic acids and the majority was gallic acid. Treatment with CRHE caused significant inhibition of AGE formation. The oral glucose tolerance test (OGTT) in normoglycemic animals showed a reduction in blood glucose levels after treatment with 100 mg/kg CHRE, accompanied by an increase in hepatic glycogen content. There was also a significant reduction in the fasting glucose levels of alloxan-induced diabetic animals after 7 days of treatment with daily doses of 100 mg/kg. After 14 weeks of hyperglycemic diet, the last four or which were combined with 100 mg/kg CRHE treatment, there was a decrease in blood triglyceride levels. There was also a statistically significant decrease in the enzymatic activity of maltase, lactase and sucrase. The results demonstrate that oral administration of 30 and 100 mg/kg CRHE inhibited AChE activity in different brain structures. Thus, the extract of C. regium showed promising antioxidant, antiglycation, and antidiabetic effects that may be related to its high phenolic content.

Anti-inflammatory action of an alkaloid, fraction and extract from Alchornea glandulosa in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Alchornea glandulosa (Euphorbiaceae) has traditionally been used in medicine for treating immune-mediated inflammatory diseases. AIM OF STUDY: This work aimed to evaluate the anti-inflammatory effects of a methanolic extract of leaves from A. glandulosa (MEAG), as well as the ethyl acetate fraction (EAFAG) and isolated compound guanidine alkaloid N-1, N-2, N-3-triisopentenylguanidine (AG-1), in experimental in vivo models of inflammation in mice. We also investigated this extract's phenols, flavonoids and flavonol compounds. MATERIALS AND METHODS: MEAG (extracted by maceration with methanol), EAFAG (fraction resulting from the partition of the methanolic extract with ethyl acetate) and AG-1 (alkaloid isolated by chromatographic methods) were analysed. MEAG and EAFAG were analysed by HPLC/DAD. The effects of MEAG (30, 100 and 300 mg/kg), EAFAG (30, 100 and 300 mg/kg) and AG-1 (5 and 30 mg/kg) were studied in the following experimental mouse models: paw oedema and myeloperoxidase (MPO) activity, croton-oil-induced ear oedema, leukocyte migration in a pleurisy model induced by carrageenan and zymosan induction of joint inflammation. RESULTS: MEAG and EAFAG were analysed by LC/DAD, and phenolic acids (gallic acid and caffeic acid) and flavonoids (myricetin-3-O-α-rhamnopyranoside and quercetin) were detected. MEAG, EAFAG and AG-1 were used in the carrageenan-induced paw oedema model and showed maximum inhibitions of 60.10% (MEAG, 2 h, 300 mg/kg) and 66.21% (EAFAG, 2 h, 300 mg/kg). AG-1 at 5 mg/kg showed significant inhibition, ranging from 60.92% to 63.13%, at all evaluated times, and the 30 mg/kg dose showed inhibition of 42.12% (1 h) and 40.36% (2 h). MEAG (37%, 46.1% and 68.11%) and EAFAG (31%, 42.21% and 48.93%), at doses of 30, 100 and 300 mg/kg, respectively, significantly reduced the increase in MPO activity, and AG-1 (5 and 30 mg/kg) showed inhibition of 64.62% and 65.12%, respectively. In the pleurisy model, MEAG (300 mg/kg), EAFAG (300 mg/kg) and AG-1 (30 mg/kg) significantly reduced the migration of total leukocytes with maximal inhibition of 80.90%, 83.17% and 89.39%, respectively. In the croton oil model, pretreatment with MEAG (0.1, 0.3 and 1 mg/ear) increased the diameter of the right ear (30.32%, 48.87% and 53.09%, respectively). Finally, MEAG (100 and 300 mg/kg; 33.11% and 56.03%) and EAFAG (100 and 300 mg/kg; 36.89% and 50.53%) reduced zymosan-induced oedema formation. CONCLUSIONS: To the best of our knowledge, these results are the first to demonstrate that A. glandulosa exhibits oral and topical anti-inflammatory activity. This study detected alkaloid and phenol/polyphenolic compounds in A. glandulosa, which may help to explain the ethnobotanical use of this plant in traditional medicine in Brazil to treat immune-mediated inflammatory diseases.

Antioxidant, anti-inflammatory, antiproliferative and antimycobacterial activities of the essential oil of Psidium guineense Sw. and spathulenol.

ETHNOPHARMACOLOGICAL RELEVANCE: Leaves from Psidium guineense Sw. are used in popular medicine for the treatment of inflammatory disease. However, there is no scientific evidence demonstrating this activity. AIM OF THE STUDY: To evaluate the antioxidant, anti-inflammatory, antiproliferative and antimycobacterial activities of the essential oil of P. guineense and spathulenol (a major constituent). The study was conducted in part to provide evidence supporting the ethnobotanical use of the leaves of this species. MATERIAL AND METHODS: The essential oil (EOPG) was extracted from the leaves of P. guineense by hydrodistillation and analysed by gas chromatography-mass spectrometry (GC-MS). The major compound, spathulenol (PG-1), was isolated in a chromatographic column and characterized by nuclear magnetic resonance (NMR). EOPG and PG-1 were evaluated in vitro for antioxidant activity by DPPH, ABTS and MDA methods; anti-inflammatory potential was assessed using two models, including pleurisy and oedema, in mice. The impact of EOPG and PG-1 on cell proliferation was determined via spectrophotometric quantification of the cellular protein content using a sulforhodamine B assay, and anti-Mycobacterium tuberculosis activity was determined using the REMA method. RESULTS: A total of 38 components were identified from the EOPG, with the sesquiterpenic alcohol spathulenol (PG-1) (80.7%) being the major constituent. EOPG and PG-1 exhibited the highest antioxidant activities in the DPPH and MDA system compared with reference standard, with IC50 values ranging from 26.13 to 85.60µg/mL. Oral administration of EOPG and PG-1 showed significant inhibition in the Cg-induced mice paw oedema and pleurisy model. The EOPG (GI50 = 0.89µg/mL) and PG-1 (GI50 = 49.30µg/mL) were particularly effective against the ovarian cancer cell line. Both showed moderate antimycobacterial activity. CONCLUSION: For the first time, this study demonstrated the antioxidant, anti-inflammatory, antiproliferative and antimycobacterial properties of the essential oil of P. guineense (leaves were collected in Dourados-MS) and spathulenol, collaborating the etnhopharmacologycal use of this plant due to its an anti-inflammatory effect.

Anti-inflammatory, antioxidant and anti-Mycobacterium tuberculosis activity of viridiflorol: The major constituent of Allophylus edulis (A. St.-Hil., A. Juss. & Cambess.) Radlk.

ETHNOPHARMACOLOGICAL RELEVANCE: The leaves of Allophylus edulis (A. St.-Hil., A. Juss. & Cambess.) Radlk. (Sapindaceae) are traditionally used as a natural anti-inflammatory agent; however, there are no scientific studies demonstrating its activity essential oil. The content of essential oil in A. edulis may be the chemical basis to explain its ethnobotanical uses, since infusions of this plant are used to treat inflammation in the traditional medicine in Brazil. AIM OF THE STUDY: This study evaluated the anti-inflammatory, antioxidant and anti-mycobacterial activities of the essential oil (EOAE) and viridiflorol, its main compound. MATERIAL AND METHODS: Essential oil from fresh leaves of A. edulis (EOAE) was obtained by hydrodistillation in a Clevenger-type apparatus. Forty-one compounds, accounting for 99.10% of the oil, were identified by gas chromatography-mass spectrometry (GC-MS). The major constituent of the oil was viridiflorol (30.88%). Additionally, the essential oil and viridiflorol were evaluated using an in vitro test against Mycobacterium tuberculosis and in 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) assays. Both EOAE (30 and 100mg/kg) and viridiflorol (3 and 30mg/kg) by oral administration were assayed in carrageenan-induced mice paw oedema and pleurisy using subcutaneous injection of dexamethasone (0.5mg/kg) as the positive control. RESULTS: EOAE and viridiflorol displayed moderate in vitro activity in the M. tuberculosis assay. In all tests, EOAE and viridiflorol showed moderate antioxidant activity compared with reference standards. Both EOAE and viridiflorol showed significant inhibition in the carrageenan-induced mice paw oedema via oral administration of the oil (30 and 100mg/kg), compound (3 and 30mg/kg), and subcutaneous injection of dexamethasone (0.5mg/kg, reference drug). Also EOAE and viridiflorol significantly inhibited carrageenan (Cg) induced pleurisy, reducing the migration of total leucocytes in mice by 62±5% (30mg/kg of oil), 35±8% (100mg/kg of oil), 71±5% (3mg/kg of viridiflorol) and 57±3% (30mg/kg of viridiflorol). CONCLUSION: For the first time, the results from this work corroborate the literature, showing that A. edulis can be used as a natural anti-inflammatory agent. Moreover, both EOAE and viridiflorol exhibited biological activities, such as anti-mycobacterial, anti-inflammatory and antioxidant activity.

Analysis of the anti-inflammatory and chemopreventive potential and description of the antimutagenic mode of action of the Annona crassiflora methanolic extract.

CONTEXT: Annona crassiflora Mart. (Annonaceae) is a medicinal plant that is widely used in folk medicine, which leads to its investigation as a potential source of new pharmacological principles. OBJECTIVE: This study describes the anti-inflammatory, antiallodynic, and antimutagenic/chemopreventive activities of the leaves A. crassiflora methanolic extract. Its antimutagenic mode of action was analyzed in a plant or animal experimental model. MATERIALS AND METHODS: Total flavonoids were quantified by spectrophotometry at 415 nm and its composition was analyzed by (1)H NMR spectra. Animals received orally, 30, 100, and 300 mg/kg of extract in both tests, carrageenan-induced paw edema and myeloperoxidase activity. Animals were treated with 100 and 300 mg/kg, in all the analyzed tests, pleural cell migration and protein exudation, carrageenan-induced cell migration into the pouch, induction of joint inflammation and carrageenan-induced allodynia response in the mouse paw. To evaluate the antimutagenic/chemopreventive activity through the Allium cepa test, we used 5, 10, and 15 mg/L of extract, and for the micronucleus test in the peripheral blood, we used the dose of 15 mg/kg. RESULTS: The fractionation of the ethyl acetate (EA) fraction, resulting from the partition of the methanol extract of the A. crassiflora, afforded through chromatographic methods resulted in the isolation of kaempferol 3-O-ß-glucoside and kaempferol 3-O-ß-diglucoside. Oral treatment with 100 and 300 mg/kg of extract significantly inhibited the carrageenan-induced edema formation, with inhibitions of 53 ± 7% and 47 ± 10%; in MPO activity, the observed inhibitions were 60 ± 7% for 100 mg/kg treatment and 63 ± 7% for 300 mg/kg. The ACME reduced significantly the total leukocytes (an inhibition of 78 ± 9% with 100 mg/kg and 90 ± 7% with 300 mg/kg) and protein levels (approximately 100% inhibition with both doses) in the pleurisy model. In carrageenan-induced leukocyte migration into the pouch, the extract inhibited leukocyte migration only when administered 300 mg/kg per dose (the reduction was 43 ± 5%). Pretreatment with extract failed to reduce the zymosan-induced edema formation and did not inhibit the carrageenan-induced mechanical allodynia. Damage reduction in Allium cepa tested with different concentrations (5, 10, and 15 mg/L) was 66.17, 75.75, and 69.19% for the pre-treatment; 72.72, 33.33, and 22.22% for the simple simultaneous treatment; 100.50, 93.93, and 102.52% for the simultaneous treatment with pre-incubation; 89.39, 79.79, and 84.34%; for the post-treatment, and 86.36, 81.31, and 93.43% for the continuous treatment. The antimutagenic evaluation in the micronucleous test showed a damage reduction of 75.00 and 64.58% for the pre-treatment and simultaneous protocols, respectively. The post-treatment protocol increased the cyclophosphamide effects in 45.83%. CONCLUSION: These results suggest that this medicinal plant has chemopreventive and anti-inflammatory therapeutic potential.

Antihyperalgesic and antidepressive actions of (R)-(+)-limonene, α-phellandrene, and essential oil from Schinus terebinthifolius fruits in a neuropathic pain model.

OBJECTIVES: Previous studies have shown that essential oil containing (R)-(+)-limonene and α-phellandrene, extracted from fruits of Schinus terebinthifolius Raddi, exhibit anti-inflammatory activity. This work aimed to verify the antihyperalgesic and antidepressive actions of (R)-(+)-limonene, α-phellandrene, and essential oil from S. terebinthifolius fruits in spared nerve injury (SNI) model of neuropathic pain in rats. METHODS: In the present work, essential oil from fruits of S. terebinthifolius, as well as the pure (R)-(+)-limonene and α-phellandrene compounds, were assayed for their effects on SNI-induced mechanical and cold hyperalgesia, and depressive-like behavior (immobility in forced swim test) in rats. The locomotor activity was evaluated in open-field test. RESULTS: Oral administration for up to 15 days of essential oil of S. terebinthifolius (100 mg/kg), (R)-(+)-limonene (10 mg/kg), α-phellandrene (10 mg/kg), and also subcutaneous 10 mg/kg dose of ketamine (positive control) significantly inhibited SNI-induced mechanical hyperalgesia and increased immobility in the forced swim test. On the 15th day of oral treatment, α-phellandrene, but neither the essential oil from S. terebinthifolius nor (R)-(+)-limonene, prevented the SNI-induced increase in sensitivity to a cold stimulus. The oral treatment with essential oil (100 mg/kg) or with compounds (10 mg/kg) did not interfere on locomotor activity. DISCUSSION: Together, the results of the present work show that essential oil of S. terebinthifolius and compounds present in this oil, including (R)-(+)-limonene and α-phellandrene, exhibit antihyperalgesic effects against mechanical hyperalgesia, and are antidepressive, while only α-phellandrene inhibited cold hyperalgesia in SNI rats.

Atividade de óleos e extratos vegetais sobre germinação carpogênica e crescimento micelial de Sclerotinia sclerotiorum / Activity of plant extracts on the carpogenic germination and mycelial growth of Sclerotinia sclerotiorum

A germinação carpogênica e o crescimento micelial de Sclerotinia sclerotiorum foram avaliados sob extratos metanólicos de Annona cacans, A. coriacea, A. crassiflora, A. dioica, A. sylvatica, Geophila repens, Guettarda viburnoides, Palicourea crocea, Schinus terebinthifolius e Trichilia silvatica, e sob as frações hexânica, hidrometanólica, clorofórmica e acetato de etila de A. cacans e óleo essencial de S. terebinthifolius. A concentração utilizada foi de 1.000 ppm para os extratos e de 100 ppm para as frações. Os extratos vegetais e as frações foram incorporados em meio ágar-água, que foi vertido em caixas gerbox com 20 escleródios. O crescimento micelial foi avaliado em óleo essencial de S. terebinthifolius, nas concentrações de 0, 100 e 1.000 ppm, incorporado ao meio BDA (Batata-Dextrose-Ágar). A germinação carpogênica apresentou-se menor sob os extratos de G. repens, P. crocea e S. terebinthifolius e sob as frações acetato de etila e clorofórmica de A. cacans. O número de apotécios formados por gerbox foi menor com o extrato de A. cacans. O crescimento micelial apresentou 10% de inibição na maior concentração do óleo essencial de S. terebinthifolius.(AU)

The efect of methanolic extracts of Annona cacans, A. coriacea, A. crassiflora, A. dioica, A. sylvatica, Geophila repens, Guettarda viburnoides, Palicourea crocea, Schinus terebinthifolius e Trichilia silvatica, and A. cacans hexane, ethyl etila, aqueous and chloroform fractions and the essential oil of S. terebinthifolius on mycelial growth and carpogenic germination of Sclerotinia sclerotiorum was evaluated. Te concentrations are 1,000 ppm for the extracts and 100 ppm for the fractions. To evaluate the germination, carpogenic, extracts and fractions were incorporated in agar-water that was poured into gerboxes where 20 sclerotia were distributed. To evaluate the mycelial growth, essential oil of S. terebinthifolius in concentrations of 0, 100 and 1,000 ppm was incorporated into the PDA and then poured into Petri dishes, to where pathogen mycelial discs were transferred. Extracts of G. repens, P. crocea and S. terebinthifolius and fractions ethyl acetate and chloroform of Annona cacans reduced the carpogenic germination of sclerotia of S. sclerotiorum and the extract of A. cacans reduced the number of apothecia formed. Mycelial growth showed 10% inhibition at the highest concentration of essential oil of S. terebinthifolius.(AU)

Potencial alelopático e antioxidante de extratos vegetais / Allelopathic and antioxidant potential of plants extracts

Objetivou-se com este trabalho avaliar a atividade alelopática, antioxidante e o conteúdo de fenóis e flavonóides totais no extrato metanólico de quatro plantas medicinais: Hibiscus sabdariffa, Ocimum gratissimum, Palicourea crocea e Trichilia silvatica. Para isso, folhas, flores e caules de Trichilia silvatica, folhas de Palicourea crocea, cálice e folhas de Hibiscus sabdariffa e folhas de Ocimum gratissimum foram utilizados. A avaliação da atividade alelopática foi realizada pelo ensaio de germinação e crescimento inicial de sementes de alface (Lactuca sativa), com três repetições de 50 sementes em delineamento inteiramente casualizado. A atividade antioxidante foi determinada pelo método fotocolorimétrico in vitro do radical livre estável 2,2-difenil-1-picril-hidrazila (DPPH). O crescimento das plântulas de alface foi mais sensível à ação dos aleloquímicos presentes nos extratos metanólicos do que a germinação das sementes, e evidenciaram o efeito inibitório dos extratos metanólicos do cálice e folhas de H. sabdariffa e das folhas de O. gratissimum e P. crocea. Os extratos metanólicos obtidos das folhas, flores e casca de T. silvatica apresentaram maior capacidade de sequestrar radicais livres com valores de IC50 de 62,36; 19,4 18,38 µg mL-1, no ensaio com DPPH e maior teor de compostos fenólicos 520,80; 545,27 e 546,56 mg de ácido gálico g de amostra-1, respectivamente, avaliado pelo ensaio de Folin-Ciocalteau.

The aim of this work was to evaluate allelopathic and antioxidant activity and total flavonoids and phenols content in methanolic extract of four medicinal plants: Hibiscus sabdariffa, Ocimum gratissimum, Palicourea crocea and Trichilia silvatica. Leaves (TSFo), flowers (TSFl) and stems (TSCa) of T. silvatica; leaves (PCFo) of P. crocea; chalice (HSCa) and leaves (HSFo) of H. sabdariffa and leaves of O. gratissimum (OGFo) were used in experiment. The evaluation of allelopathic activity was carried out by test of seed germination and initial growth of lettuce seedlings (Lactuca sativa), with three replications of 50 seeds in completely randomized design. The antioxidant activity was determined by method photo colorimetric in vitro of stable free radical 2.2-diphenyl-1-picrylhydrazyl (DPPH). The growth of lettuce seedlings was more sensitive to the action of allelochemicals in methanolic extracts than seeds germination, and showed the inhibitory effect of methanolic extracts from leaves and chalice of H. sabdariffa and leaves of O. gratissimum and P. crocea. Methanolic extracts obtained from the leaves (TSFo), flowers (TSFl) and bark (TSCa) of T. silvatica showed higher ability to scavenging free radical with IC50 values of 62.36; 19.4 18.38 µg mL-1, in assay with DPPH and highest content of phenolics, 520.80; 545.27 546.56 mg gallic acid per g of sample, respectively, evaluated by the Folin-Ciocalteau assay.

Potencial alelopático de Tropaeolum majus L. na germinação e crescimento inicial de plântulas de picão-preto / Allelophaty potential of Tropaeolum majus L on picão-preto seeds germination and initial seedling growth

Objetivou-se com este estudo avaliar o potencial alelopático de extratos metanólicos de folhas, flores e raízes de capuchinha (Tropaeolum majus L.) sobre a germinação de sementes e o crescimento inicial de plântulas de picão-preto. O extrato metanólico com melhor potencial de inibição foi submetido a particionamento, resultando nas frações hexânica, clorofórmica, acetato de etila e hidrometanólica e posterior caracterização pelo espectro de absorção na região do infravermelho (IV). O efeito alelopático foi avaliado sobre as sementes de picão-preto, as quais foram distribuídas sobre papel germitest umedecido com 2mL dos extratos e mantidas em germinador do tipo B.O.D. regulado a temperatura de 25°C e luz branca constante, sendo que as sementes imersas diretamente em água constituíram o tratamento controle. A avaliação da qualidade da semente foi realizada pelos testes de germinação e vigor (primeira contagem e comprimento de raiz primária e de hipocótilo das plântulas), em delineamento inteiramente ao acaso. O potencial alelopático das folhas de capuchinha foi maior em relação às demais partes da planta sobre a germinação das sementes, comprimento de hipocótilo e de raiz das plântulas de picão-preto. Estes efeitos podem estar associados à presença de grupos químicos polares, pois à medida que se aumentou a polaridade dos solventes detectou-se maior efeito inibitório sobre a germinação e o crescimento inicial de plântulas de picão-preto.

This research aimed to evaluate the metanolic extracts allelopathic potential from leaves, flowers and roots of capuchinha (Tropaeolum majus L.) on picão-preto seeds germination and initial seedling growth. The best inhibitor metanolic extract was fractioned, in hexanic, cloroformic, etil acetate and hidrometanolic fractions and it was characterized through absorption spectrum using mid-infrared. To evaluate the allelopathic effect of metanolic extracts and the fractions aftereffect; it was used picão-preto seeds distributed on 2ml extracts moistured paper maintained in B.O.D. germinator at 25°C and white constant light. Seeds sowed directly in water moistured germitest paper constituted the control. The allelopathic potential extracts were evaluated through the first count, final germination percentage, seedlings primary root and hypocotyl length (cm). The experiment was carried out DIC with three replications of 25 seeds. The capuchinha leaves showed allelopathic potential on picão-preto seeds germination, hypocotyl and seedlings primary root length. It was suggested that such effects are associated to the presence of polar chemical groups because the increase in the solvents polarity detected the major picão-preto germination and seedlings growth inhibition effect.