Evaluation of acute toxicity, genotoxicity and inhibitory effect on acute inflammation of an ethanol extract of Morus alba L. (Moraceae) in mice.
BACKGROUND
Morus alba L. (white mulberry) is used in traditional medicine worldwide, including Brazil. The leaves of this plant are used to treat inflammatory disorders. Universal interest in this plant necessitates studies on the toxicological safety and scientific substantiation of the medicinal properties of M. alba. In previous work, we investigated the acute toxicity of orally administered M. alba ethanol extract in mice.


OBJECTIVE
This work was designed to investigate the ethanol extract obtained from M. alba leaves for acute toxicity when intraperitoneally administered, in vivo genotoxicity, and potential to reduce acute inflammation. In order to further investigate the constituents of the extract, we also obtained the high-performance liquid chromatography (HPLC) fingerprint of the extract.


METHODS
Phytochemical analysis by thin layer chromatography (TLC) was performed and the results were used to obtain the HPLC fingerprint. Acute toxicity of 300 and 2000mg/kg b.w. i.p. doses administered to mice for 14 days was evaluated. Genotoxicity was evaluated by counting the number of micronucleated polychromatic erythrocytes in the blood of mice that either received or did not receive the extract at 75, 150 and 300mg/kg b.w. per os. The anti-inflammatory effect of the same doses administered per os was investigated using the carrageenan air pouch model.


RESULTS
The TLC analysis of the extract revealed the presence of a remarkable amount of flavonoids and cinnamic acids. The HPLC fingerprint showed the presence of one major peak corresponding to chlorogenic acid and two smaller peaks corresponding to flavonoids. In the toxicity assays, there were no deaths or deviations in behavior of treated mice as compared to the control at any dose. However, biochemical, hematological, and histological analyses showed that intraperitoneal injection caused several forms of damage to the mice, which were not observed in case of oral administration, studied in our previous work. Oral administration of the extract did not result in genotoxicity and considerably reduced (58.6-65.6% inhibition) leukocyte migration in all doses evaluated, in comparison with the negative control.


CONCLUSIONS
The ethanol extract from M. alba leaves administered intraperitoneally possesses a greater degree of toxicity in mice when compared to per os administration. The extract was not genotoxic when ingested by mice and exhibited a highly inhibitory effect against acute inflammation, which is probably linked to the presence of chlorogenic acid and flavonoids in the composition. This work contributes to the determination of safety of the medicinal use of M. alba leaves.
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Alisson Macário de
AM
Oliveira
Departamento de Antibióticos, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Matheus Ferreira do
MF
Nascimento
Departamento de Farmácia, Centro de Ciências da Saúde, Universidade Federal de Pernambuco, 50740-520 Recife, Pernambuco, Brazil.
Magda Rhayanny Assunção
MR
Ferreira
Centro Acadêmico de Vitória, Universidade Federal de Pernambuco, 55604-000 Vitória de Santo Antão, Pernambuco, Brazil.
Danielle Feijó de
DF
Moura
Centro Acadêmico de Vitória, Universidade Federal de Pernambuco, 55604-000 Vitória de Santo Antão, Pernambuco, Brazil.
Talita Giselly Dos Santos
TG
Souza
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Gabriela Cavalcante da
GC
Silva
Departamento de Antibióticos, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Eduardo Henrique da Silva
EH
Ramos
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Patrícia Maria Guedes
PM
Paiva
Departamento de Histologia e Embriologia, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420, Recife, Pernambuco, Brazil.
Paloma Lys de
PL
Medeiros
Departamento de Antibióticos, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Teresinha Gonçalves da
TG
Silva
Departamento de Farmácia, Centro de Ciências da Saúde, Universidade Federal de Pernambuco, 50740-520 Recife, Pernambuco, Brazil.
Luiz Alberto Lira
LA
Soares
Centro Acadêmico de Vitória, Universidade Federal de Pernambuco, 55604-000 Vitória de Santo Antão, Pernambuco, Brazil.
Cristiano Aparecido
CA
Chagas
Departamento de Antibióticos, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil.
Ivone Antônia de
IA
Souza
Departamento de Bioquímica, Centro de Biociências, Universidade Federal de Pernambuco, 50670-420 Recife, Pernambuco, Brazil. Electronic address: thiagohn86@yahoo.com.br.
Thiago Henrique
TH
Napoleão
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D000431:Ethanol / Q000737:chemistry; D005260:Female; D007249:Inflammation / Q000517:prevention & control*; D008297:Male; D051379:Mice; D031625:Morus / Q000737:chemistry*; D009152:Mutagenicity Tests; D010936:Plant Extracts / Q000494:pharmacology* / Q000633:toxicity; D023382:Toxicity Tests, Acute
D010936:Plant Extracts; D000431:Ethanol
Anti-inflammatory effect; Micronucleus test; Morus alba; Toxicological safety; White mulberry