Antigenotoxicity properties of Copaifera multijuga oleoresin and its chemical marker, the diterpene (−)-copalic acid

• Published in: Journal of Toxicology and Environmental Health, Part A Vol. 81; no. 5; pp. 116 - 129
• Main Authors: Alves, Jacqueline Morais, Leandro, Luís Fernando, Senedese, Juliana Marques, Castro, Pâmela Tinti de, Pereira, Daiane Eleutério, Resende, Flávia Aparecida, Campos, Débora Leite, Silva, Jonas Joaquim Mangabeira da, Varanda, Eliana Aparecida, Bastos, Jairo Kenupp, Ambrósio, Sérgio Ricardo, Tavares, Denise Crispim
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01-01-2018; Taylor & Francis Ltd
• Subjects: (−)-copalic acid; 2-Aminofluorene; Abbreviations; Acids; Aflatoxin B1; Agricultural research; Ames test; Animals; antigenotoxicity; Antimutagenic Agents - pharmacology; Audio frequencies; Benzo(a)pyrene; Benzo(a)pyrene-diol epoxide; Bioassays; Biochemical oxygen demand; Chromatography; Comet Assay; Copaifera multijuga; Copaifera multijuga Hayne; Cricetulus; Damage detection; Diterpenes - pharmacology; Erythrocytes; Fabaceae - chemistry; Fibroblasts - drug effects; Gas chromatography; Genotoxicity; High performance liquid chromatography; In vitro methods and tests; In vivo methods and tests; Inspection; Liquid chromatography; Lung; Lungs; Male; Mass spectrometry; Mass spectroscopy; Methyl methanesulfonate; Mice; micronucleus test; Micronucleus Tests; Mitomycin C; Mutagenicity; Mutagenicity Tests; Mutagens; Natural products; NMR; Nondestructive testing; Nuclear magnetic resonance; o-Phenylenediamine; Oxygen demand; Phenylenediamine; Phosphates; Plant Extracts - pharmacology; Pyrene; Revertants; Scientific imaging; Sodium; Sodium azide; Spectroscopy; Toxicity; genotoxicity; Ames test; comet assay; antigenotoxicity; micronucleus test; (−)-copalic acid; Copaifera multijuga Hayne
• ISSN: 1528-7394; 1087-2620; 2381-3504
• DOI: 10.1080/15287394.2017.1420505

In view of the biological activities and growing therapeutic interest in oleoresin obtained from Copaifera multijuga, this study aimed to determine the genotoxic and antigenotoxic potential of this oleoresin (CMO) and its chemical marker, diterpene (−)-copalic acid (CA). The micronucleus (MN) assay in V79 cell cultures and the Ames test were used for in vitro analyses, as well as MN and comet assays in Swiss mice for in vivo analyses. The in vitro genotoxicity/mutagenicity results showed that either CMO (30, 60, or 120 µg/ml-MN assay; 0.39-3.12 mg/plate-Ames test) or CA (2.42; 4.84, or 9.7 µg/ml-MN assay; 0.39-3.12 mg/plate-Ames test) did not induce a significant effect on the frequency of MN and number of revertants, demonstrating an absence of genotoxic and mutagenic activities, respectively, in vitro. In contrast, these natural products significantly reduced the frequency of MN induced by methyl methanesulfonate (MMS), and exerted a marked inhibitory effect against indirect-acting mutagens in the Ames test. In the in vivo test system, animals treated with CMO (6.25 mg/kg b.w.) exhibited a significant decrease in rate of MN occurrence compared to those treated only with MMS. An antigenotoxic effect of CA was noted in the MN test (1 and 2 mg/kg b.w.) and the comet assay (0.5 mg/kg b.w.). Data suggest that the chemical marker of the genus Copaifera, CA, may partially be responsible for the observed chemopreventive effect attributed to CMO exposure.
Abbreviations: 2-AA, 2-anthramine; 2-AF, 2-aminofluorene; AFB
1
, aflatoxin B
1
; B[a]P, benzo[a]pyrene; BOD, biological oxygen demand; BPDE, benzo[a]pyrene-7,8-diol-9,10-epoxide; CA, (−)-copalic acid; CMO, oleoresin of Copaifera multijuga, DMEM, Dulbecco`s Modified Eagles`s Medium; DMSO, dimethylsulfoxide; EMBRAPA, Brazilian agricultural research corporation; GC-MS, gas chromatography-mass spectrometry; HAM-F10, nutrient mixture F-10 Ham; HPLC, high performance liquid chromatography; LC-MS, liquid chromatography-mass spectrometry; MI, mutagenic index; MMC, mitomycin C; MMS, methyl methanesulfonate; MN, micronucleus; MNPCE, micronucleated polychromatic erythrocyte; NCE, normochromatic erythrocyte; NDI, nuclear division index; NMR, nuclear magnetic resonance; NPD, 4-nitro-o-phenylenediamine; PBS, phosphate-buffered saline; PCE, polychromatic erythrocyte; SA, sodium azide; V79, Chinese hamster lung fibroblast.