title: Analysis of the anti-inflammatory and chemopreventive potential and description of the antimutagenic mode of action of the Annona crassiflora methanolic extract.
abstract: BACKGROUND
  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.
  
  
  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-β-glucoside and kaempferol 3-O-β-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%.
  
  
  CONCLUSIONS
  These results suggest that this medicinal plant has chemopreventive and anti-inflammatory therapeutic potential.
authors.affiliation: a Center of Studies in Stem Cells, Cell Therapy and Genetic Toxicology (CeTroGen), University Hospital (NHU), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil .|b Center of Biological and Health Sciences (CCBS), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil .
authors.forename: Roberta Schroder
authors.initials: RS
authors.lastname: Rocha
authors.affiliation: c Faculty of Health Sciences (FCS), Federal University of Grande Dourados (UFGD) , Dourados , MS , Brazil .
authors.forename: Candida Aparecida Leite
authors.initials: CA
authors.lastname: Kassuya
authors.affiliation: d Faculty of Agricultural Sciences (FCA), Federal University of Grande Dourados (UFGD) , Dourados , MS , Brazil .
authors.forename: Anelise Samara Nazari
authors.initials: AS
authors.lastname: Formagio
authors.affiliation: a Center of Studies in Stem Cells, Cell Therapy and Genetic Toxicology (CeTroGen), University Hospital (NHU), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil .|e Midwest Pro Network - Graduate Program in Biotechnology and Biodiversity, Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil .
authors.forename: Mariana de Oliveira
authors.initials: Mde O
authors.lastname: Mauro
authors.affiliation: c Faculty of Health Sciences (FCS), Federal University of Grande Dourados (UFGD) , Dourados , MS , Brazil .
authors.forename: Magaiver
authors.initials: M
authors.lastname: Andrade-Silva
authors.affiliation: b Center of Biological and Health Sciences (CCBS), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil .
authors.forename: Antonio Carlos Duenhas
authors.initials: AC
authors.lastname: Monreal
authors.affiliation: b Center of Biological and Health Sciences (CCBS), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil .|f Master's Programme in Pharmacy, Center of Biological and Health Sciences (CCBS), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil , and.
authors.forename: Andréa Luiza
authors.initials: AL
authors.lastname: Cunha-Laura
authors.affiliation: d Faculty of Agricultural Sciences (FCA), Federal University of Grande Dourados (UFGD) , Dourados , MS , Brazil .
authors.forename: Maria do Carmo
authors.initials: Mdo C
authors.lastname: Vieira
authors.affiliation: a Center of Studies in Stem Cells, Cell Therapy and Genetic Toxicology (CeTroGen), University Hospital (NHU), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil .|f Master's Programme in Pharmacy, Center of Biological and Health Sciences (CCBS), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil , and.|g Graduate Program in Health and Development in the Midwest Region, School of Medicine "Dr. Hélio Mandetta" (FAMED), Federal University of Mato Grosso do Sul (UFMS) , Campo Grande , MS , Brazil.
authors.forename: Rodrigo Juliano
authors.initials: RJ
authors.lastname: Oliveira
mesh_terms: D000700:Analgesics / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D000818:Animals; D031041:Annona / Q000737:chemistry*; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D016588:Anticarcinogenic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D016587:Antimutagenic Agents / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D002351:Carrageenan; D002634:Chemotaxis, Leukocyte / Q000187:drug effects; D004195:Disease Models, Animal; D004305:Dose-Response Relationship, Drug; D005260:Female; D006930:Hyperalgesia / Q000139:chemically induced / Q000517:prevention & control*; D007249:Inflammation / Q000139:chemically induced / Q000276:immunology / Q000517:prevention & control*; D008297:Male; D000432:Methanol / Q000737:chemistry*; D051379:Mice; D048629:Micronuclei, Chromosome-Defective / Q000187:drug effects*; D015162:Micronucleus Tests; D008517:Phytotherapy; D010936:Plant Extracts / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*; D018515:Plant Leaves; D010946:Plants, Medicinal; D066244:Proton Magnetic Resonance Spectroscopy; D012997:Solvents / Q000737:chemistry*; D013997:Time Factors
chemical_list: D000700:Analgesics; D000893:Anti-Inflammatory Agents; D016588:Anticarcinogenic Agents; D016587:Antimutagenic Agents; D010936:Plant Extracts; D012997:Solvents; D002351:Carrageenan; D000432:Methanol
keywords: Allium cepa; antimutagenesis; carrageenan; micronucleus; paw edema