Mice Behavioral Phenotype Changes after Administration of Anani (Symphonia globulifera, Clusiaceae), an Alternative Latin American and African Medicine

Abstract

Background: Anani, (Symphonia globulifera, Clusiaceae), known as chewstick, is a traditional plant occurring in Africa and in Central and South Americas that is used against parasites and microorganisms. Although its use is popular in some of these countries, there is a lack of information related to its influence over behavioral phenotype (BP).

Objective: The objective of this study is to evaluate the influence of the administration of the extract obtained from the aerial organs of Anani (EB1257) to male Balb-c mice over BP.

Materials and methods: Open cage observation, open field, and elevated-plus maze apparatuses were used. Evaluations were done 15, 30, 60, 120, and 180 min after intraperitoneal administration of Anani extract.

Results: Impairment of general behavior activity, response to touch, tail squeeze, defecation, locomotion and rearing frequency were observed although no signs of hemorrhage or macroscopical alterations of internal organs. Anani is harmful, but not toxic if used in the appropriate doses, yet to be determined to male mice. Impairment of locomotion and defecation was observed, indicating some degree of influence over locomotion, but no alterations in anxiety levels were assessed. Three compounds were previously found in the plant-lupeol (1), β-amyrin (2) and 3-β-hydroxyglutin-5-ene (3), and one is being described for the first time to occur in the species: oleanolic acid (4).

Conclusions: The present work contributes in the support of the rational use of Anani, an important Latin American and African alternative medicine, presenting findings that are being reported for the first time.

Summary: Symphonia globulifera impairs locomotion and defecatin in behavior analysesNo alterations in anxiety was observedOleanolic acid occurs in the species. Abbreviations used: BP: Behavioral phenotype; OF: Open field, EPM: Elevated-plus maze, MMA/ICMBio/SISBIO: Ministério do Meio Ambiente/Instituto Chico Mendes de Conservação da Biodiversidade/Sistema de Autorização e Informação em Biodiversidade, IBAMA/MMA/CGen: Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis/Ministério do Meio Ambiente/Conselho de Gestão do Patrimônio Genético, AM: Amazonas State, UNIP: Universidade Paulista, mg: milligram, kg: kilogram, I.P: Intraperitoneal, CEUA/ICS/UNIP: Comissão de Ética no Uso de Animais/Instituto de Ciências da Saúde/Universidade Paulista, LD: Lethal dose, NLD: Nonlethal dose, GBA: General behavior activity, FCHCL₃: Fraction chloroform, FBuOH: Fraction buthanol, FH₂O: Fraction water, FrHEX: Fraction hexane, FrDCM: Fraction dichloromethane, FrMeOH: Fraction methanol, ¹³C NMR: Carbon nuclear magnetic resonance, EPA: United States Environmental Protection Agency.

Keywords: Behavioral phenotype; Symphonia globulifera; general activity; locomotion.

Figure 1

Experimental design diagrams. Behavioral effects of EB1257 intraperitonealy injected in mice: flowchart and timeline related to the experimental design of the experiments. GBA: General behavior activity; OF: Open field; EPM: Elevated-plus maze

Figure 2

Compounds isolated from Symphonia globulifera organic extract. Lupeol (1), β-amyrin (2), 3-β-hydroxyglutin-5-ene (3), and oleanolic acid (4)

Figure 3

Behavior alterations observed in male Balb-c mice after administration of organic extract EB1257, obtained from Symphonia globulifera (Clusiaceae), in stage 1 of the experiment. the parameters that were observed was (a) General behavior; (b) Response to touch; (c) Tail squeeze; (d) Corneal reflex; (e) ataxia and (f) defecation. Kruskal-Wallis nonparametric variance test was performed (n = 3; n_total = 12), followed by the Dunnet posttest, median ± range is represented; significance among medians was considered if P < 0.05. For defecation analysis, a parametric one-way ANOVA was performed (n = 3; n_total = 12), followed by Bonferroni posttest, mean ± standard error is represented and significance among means was achieved if P < 0.05. *=P<0.05, **=P<0.01, ***P<0.001

Figure 4

Behavior effects over parameters that were influenced by the intraperitoneal administration of nonlethal dose of organic extract EB1257, obtained from Symphonia globulifera (Clusiaceae), in male Balb-c mice. The following parameters were observed in stage 2; (a) General behavior; (b) Response to touch; (c) Tail squeeze; (d) ptosis; (e) Defecation and (f) Cyanosis. Mann-Whitney test was performed (P < 0.05). VC = Vehicle control. *=P<0.05, **=P<0.01

Figure 5

Results related to open field analysis in the second stage of the experiment, after intraperitoneal administration of the nonlethal dose of organic extract, obtained from Symphonia globulifera (Clusiaceae), to male Balb-c mice. The following parameters were observed in stage 2: a. locomotion frequency; b. immobility time; c. rearing frequency; d. Defecation; e. Grooming. Data are are presented as means ± standard error (n = 10; n_total = 20). Two-way repeated measures ANOVA followed by Bonferroni posttest were used in the analyses. Significance was observed if P < 0.05. *=P<0.05, **=P<0.01, ***P<0.001