Natural killer activity of the spleen cells of Ehrlich tumor-bearing mice treated with Copaifera multijuga extract

Paloma Filomena Gouveia Rodrigues, Lucinéia Reuse Albiero, Eduardo Figueredo Nery, Taiany Oliveira Kelly, Jeniffer Charlene Silva Dalazen, Débora Linsbinski Pereira, Adilson Paulo Sinhorin, Valéria Dornelles Gindri Sinhorin, Lindsey Castoldi


AIMS: Copaifera multijuga Hayne oleoresin is commonly used in traditional medicine owing to its anti-inflammatory, antiseptic, antitumor, and antibacterial properties. However, little is known about the effect of the compounds from the bark of this plant. In this study, the immunomodulatory effect of the ethanolic extract of C. multijuga bark via natural killer activity of non-adherent spleen cells of Ehrlich tumor-bearing mice was evaluated.

METHODS: Male Swiss mice were inoculated subcutaneously with 1×106 Ehrlich tumor cells (Ehrlich and Ehrlich/C. multijuga group) or phosphate buffered saline solution (control group and C. multijuga group) and treated orally daily with C. multijuga extract (200 mg kg-1, 0.1 mL per mouse, for the Ehrlich/C. multijuga and C. multijuga groups) or phosphate buffered saline solution (control group and Ehrlich group). The four experimental groups consisted in eight mice each and were organized in two sets, one treated for seven days and another treated for 14 days, totalizing 64 mice throughout the experiment. Twenty-four hours after the last oral administration, the mice were euthanized and the spleen tissue was isolated to prepare a non-adherent spleen cell suspension and to evaluate natural killer activity. Data are presented as the cell lysis percentage of Yac.1 target cells by non-adherent spleen cells.

RESULTS: Treatment for seven days increased natural killer activity in the Ehrlich/C. multijuga group (21.20±8.89, p<0.05) compared to the control group (3.14±2.71, p<0.05); however, this effect was not maintained in the groups treated for 14 days (Control: 6.02±6.98, Ehrlich: 4.82±7.72, C. multijuga: 2.07±2.10, Ehrlich/C. multijuga: 2.01±1.63, p>0.05).

CONCLUSIONS: Treatment for seven days with an ethanolic extract of C. multijuga bark enhanced the natural killer activity of non-adherent spleen cells from Ehrlich tumor-bearing mice.


Copaifera multijuga; fabaceae; copaiba; natural killer cells; carcinoma, Ehrlich tumor; immunomodulation.

Full Text:



Tobouti PL, Martins TCA, Pereira TJ, Mussi MCM. Antimicrobial activity of copaíba oil: A review and a call for further research. Biomed Pharmacother. 2017;94:93-9.

Ricardo LM, Dias BM, Mügge FLB, Leite VV, Brandão MGL. Evidence of traditionality of Brzilian medicinal plants: The case studies of Stryphnodendron adstringens (Mart.) Coville (barbatimão) barks and Copaifera spp. (copaíba) oleoresin in wound healing. J Ethnopharmacol. 2018;219:319-36.

Sultana S, Asif HM, Nazar HMI, Akhtar N, Rehman JU, Rehman RU. Medicinal Plants Combating Against Cancer – a Green Anticancer Approach. Asian Pac J Cancer Prev. 2014;15(11):4385-94.

BRASIL. Lei n° 71, de 2 de setembro de 2013. Diário Oficial da República, Brasília, DF, 2 set. 2013. Série 1, p. 5439.

Albuquerque KCO, Veiga ASS, Silva JVS, Brigido HPC, Ferreira EPR, Costa EVS, Marinho AMR, Percário S, Dolabela M.F. Brazilian Amazon Traditional Medicine and the Treatment of Difficult to Heal Leishmaniasis Wounds with Copaifera. Evid Based Complement Alternat Med. 2017;2017:8350320.

Trindade R, Silva JK, Setzer WN. Copaifera of the Neotropics: A Review of the Phytochemistry and Pharmacology. Int J Mol Sci. 2018;19(5):1511-44.

Diefenbach AL, Muniz FWMG, Oballe HJR, Rosing CK. Antimicrobial activity of copaíba oil (Copaifera ssp.) on oral pathogens: Systematic review. Phytother Res. 2018;32:586-96.

Ghizoni CVC, Ames APA, Lameira OA, Amado CAB, Nakanishi ABS, Bracht L, Natali MRM, Peralta RM, Bracht A, Comar JF. Anti-inflammatory and Antioxidant Actions of Copaíba Oil Are Related to Liver Cell Modifications in Arthritic Rats. J Cell Biochem. 2017;118:3409-23.

Albiero LR, Nery EF, Dalazen JC, Kelly TO, Pereira DL, Sinhorin VDG, Kaneno R, Castoldi L. Ethanolic extracts of Copaifera Multijuga Inhibits the Subcutaneous Growth of Ehrlich Carcinoma in Swiss Mice. IOSR-JPBS. 2016;11(5):30-8.

Cassali GD, Silva AE, Santos FGA. Marcadores de proliferação celular na avaliação do crescimento do tumor sólido e ascítico de Ehrlich. Arq Bras Med Vet Zootec. 2006;58:658-61.

Calixto-Campos C, Zarpelon AC, Corrêa M, Cardoso RDR, Pinho-Ribeiro FA, Cecchini R, Moreira EG, Crespigio J, Bernardy CCF, Casagrande R, Verri Jr W. The Ehrlich’s tumor induces pain-like behavior in mice: a novel model of cancer pain for pathophysiological studies and pharmacological screening. Biomed Res Int. 2013;2013:624815.

Frajacomo FTT, Padilha CS, Marinello PC, Guarnier FA, Cecchini R, Duarte JAR, Deminice R. Solid Ehrlich carcinoma reproduces functional and biological characteristics of cancer cachexia. Life Sci. 2016;162:47-53.

Valadares MC, Klein SI, Guaraldo AMA, Queiroz MLS. Enhancement of natural killer cell function by titanocenes in mice bearing Ehrlich ascites tumour. Eur J Pharmacol. 2003;473:191-6.

Pratheeshkumar P, Kuttan G. Effect of vernolide-A, a sesquiterpene lactone from Vernonia cinereal L., on cell-mediated immune response in B16F-10 metastatic melanoma-bearing mice. Immunopharmacol Immunotoxicol. 2011;33(3):533-8.

Houh YK, Kim KE, Park S, Hur DY, Kim S, Kim D, Bang SI, Yang Y, Park HJ, Cho D. The Effects of Artemisinin on the Cytolytic Activity of Natural Killer (NK) Cells. Int J Mol Sci. 2017;18(7):1600-11.

Koch J, Steinle A, Watzl C, Mandelboim O. Activiting natural cytotoxicity receptors of natural killer cells in cancer and infection. Trends Immunol. 2013;34(4):182-91.

Pratheeshkumar P, Kuttan G. Modulation of cytotoxic T lymphocyte, natural killer cell, antibody-dependent cellular cytotoxicity, and antibody-dependent complement-mediated cytototxicity by Vernonia cinereal L. and vernolide-A in BALB/c mice via enhanced production of cytokines IL-2 and IFN-Immunopharmacol Immunotoxicol. 2012;34(1):46-55.

Silva SL, Figueiredo PMS, Yano T. Chemotherapeutic potencial of the volatile oils from Zanthoxylum rhoifolium Lam leaves. Eur J Pharmacol. 2007;576:180-8.

Cunha APS, Baldissera L, Pereira DL, Albiero LR, Castoldi L, Sinhorin AP, Sinhorin VDG. Evaluation of the antioxidant potencial of Copaifera multijuga in Ehrlich tumor-bearing mice. Acta Amazon. 2019;49(1):41-7.

Botelho NM, Corrêa SC, Lobato RC, Teixeira RKC, Quaresma JAS. Immunohistochemistry of the uterine cervix of rats bearing the Walker 256 tumor treated with copaíba balsam. Acta Cir Bras. 2013;28(3):185-9.

Dias FGG, Jorge AT, Pereira LF, Furtado RA, Ambrósio SR, Bastos JK, Ramos SB, Chahud F, Dias LGGG, Honsho CS, Tavares DC. Use of Copaifera multijuga for acute corneal repair after chemical injury: A clinical, histopathological and toxicogenetic study. Biomed Pharmacother. 2017;96:1193-98.

Chen L, Wei Y, Zhao S, Zhang M, Yan X, Gao X, Li J, Gao Y, Zhang A, Gao Y. Antitumor and immunomodulatory activities of total flavonoids extract from persimmon leaves in H22 liver tumor-bearing mice. Sci Rep. 2018;8:10523-34.

Holderness J, Jackiw L, Kimmel E, Kerns H, Radke M, Hedges JF, Petrie C, McCurley P, Glee PM, Palecanda A, Jutila MA. Select Plant Tannins Induce IL-2RUp-Regulation and Augment Cell Division in  T Cells. J Immunol. 2007;179:6468-78.

Lisanti A, Formica V, Ianni F, Albertini B, Marinozzi M, Sardella R, Natalini B. Antioxidant activity of phenolic extracts from different cultivars of Italian onion (Allium cepa) and relative human immune cell proliferative induction. Pharm Biol. 2016;54(5):799-806.

Pereira DL, da Cunha APS, Cardoso CRP, da Rocha CQ, Vilegas W, Sinhorin AP, Sinhorin VDG. Antioxidant and hepatoprotective effects of ethanolic and ethyl acetate stem bark extracts from Copaifera multijuga (Fabaceae) in mice. Acta Amazon. 2018;48(4):348-58.

Brito NMB, Brito MVH, Carvalho RKV, Moura LTM, Matos B, Lobato RC, Correa SC, Brito RB. The effect of copaíba balsam on Walker 256 carcinoma inoculated into the vagina and uterine cervix of female rats. Acta Cir Bras. 2010;25:176-80.

Lucca LG, Matos SP, Kreutz T, Teixeira HF, VeigaJr VF, Araújo BV, Limberger RP, Koester LS. Anti-inflammatory effetct from a hydrogel containing nanoemulsified copaíba oil (Copaifera multijuga Hayne). AAPS PharmSciTech. 2017;19(2):522-30.

Furtado RA, Oliveira PF, Senedese JM, Ozelin SD, Souza LDR, Leandro LF, Oliveira WL, Silva JJM, Oliveira LC, Rogez H, Ambrosio SR, Veneziani RCS, Bastos JK, Tavares DC. Assessment of genotoxic activity of oleoresins and leaves extracts of six Copaifera species for prediction of potencial human risks. J Ethnopharmacol. 2018;221:119-25.

Alves JM, Senedese JM, Leandro LF, Castro PT, Pereira DE, Carneiro LJ, Ambrósio SR, Bastos JK, Tavares DC. Copaifera multijuga oleoresin and its constituent diterpene (-)-copalic acid: Genotoxicity and chemoprevention study. Mutat Res Gen Tox En. 2017;819:26-30.


This journal is a member of COPE (Committee on Publication Ethics) and follow the principles recommended by this international reference organization on integrity and ethics in scientific publication.

Licença Creative Commons
Except where otherwise noted, the material published in this journal is licensed under a Creative Commons Attribution 4.0 International licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original publication is properly cited.
Attribution — You must give appropriate credit, provide a link to the license, and indicate if changes were made. You may do so in any reasonable manner, but not in any way that suggests the licensor endorses you or your use.

 Member of OASPA

Copyright: © 2006-2019 EDIPUCRS