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Avalia??o da atividade anti-inflamat?ria do extrato aquoso, fra??es e compostos identificados nos frutos de Hancornia Speciosa Gomes (Apocynaceae)R?go, Manoela Torres do 19 June 2015 (has links)
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Previous issue date: 2015-06-19 / Hancornia speciosa Gomes (Apocynaceae), conhecida popularmente
por ?mangabeira?, ? usada na medicina popular para o tratamento de desordens
inflamat?rias, hipertens?o, dermatites, diabetes, doen?as hep?ticas e
desordens estomacais. No que diz respeito aos frutos de H. speciosa, a
etnobot?nica indica o uso principalmente para o tratamento de inflama??o e
tuberculose, entretanto n?o h? relatos de estudos que comprovem sua poss?vel
atividade biol?gica. O estudo tem como objetivo avaliar a atividade antiinflamat?ria
do extrato aquoso dos frutos de H. speciosa Gomes. O extrato
aquoso foi preparado por decoc??o, posteriormente submetido a um
fracionamento por meio de parti??o l?quido-l?quido e os metabolitos
secund?rios identificados por Cromatografia l?quida de alta efici?ncia acoplada
ao detector de arranjos de diodos (CLAE-DAD) e Cromatografia l?quida
acoplada ao detector de arranjo de diodo e ao espectr?metro de massa (CLDAD-EM).
As propriedades anti-inflamat?rias do extrato aquoso, fra??es
diclorometano (CH2Cl2), acetato de etila (AcOEt) e n-butanol (n-BuOH), bem
como a rutina e o ?cido clorog?nico foram investigadas usando modelos in vivo
e in vitro. Testes in vivo compreenderam edema de orelha induzido por xilol,
onde foi mensurado a forma??o do edema; peritonite induzida por carragenina
onde foi avaliado os leuc?citos totais em 4h e bolsa de ar induzida por zimosam
que foi mensurado a contagem total de leuc?citos e contagem diferencial em 6,
24 e 48 horas. Os testes in vitro avaliaram os n?veis de citocinas IL-1?, IL-6, IL-
12 e TNF-? usando ELISA a partir do modelo de peritonite induzido por
carragenina. Os resultados demonstraram a presen?a de rutina e ?cido
clorog?nico no extrato aquoso dos frutos de H. speciosa por CLAE-DAD e CLDAD-EM.
O extrato aquoso e fra??es, bem como a rutina e ?cido clorog?nico
inibiram significativamente o edema de orelha induzido por xilol e reduziram a
migra??o celular em modelos animais, tanto na peritonite induzida por
carragenina como na bolsa de ar induzida por zimosam. Al?m disso, foi
observada a redu??o dos n?veis de express?o de citocinas IL-1?, IL-6, IL-12 e
TNF-?. Este ? o primeiro estudo que demonstra o efeito anti-inflamat?rio do
extrato aquoso dos frutos de Hancornia speciosa contra diferentes agentes
inflamat?rios em modelos animais, sugerindo que suas mol?culas bioativas,
especialmente rutina e ?cido clorog?nico contribuem, em parte, para o efeito
anti-inflamat?rio do extrato aquoso. Estes resultados suportam a utiliza??o de
H. speciosa na medicina popular e revelam que o extrato aquoso apresenta um
potencial terap?utico para o desenvolvimento de uma droga herbal com
propriedades anti-inflamat?rias. / Hancornia speciosa Gomes (Apocynaceae), popularly known as
?mangabeira?, has been used in folk medicine to treat inflammatory disorders,
hypertension, dermatitis, diabetes, liver diseases and stomach disorders.
Regarding the Hancornia speciosa fruits, the ethnobotany indicates its use
especially for treating inflammation and tuberculosis. However, no study has
been done so far to prove such biological activities. The objective was
evaluation anti-inflammatory activity from the fruits of Hancornia speciosa
Gomes (mangabeira). Aqueous extract was prepared by decoction,
subsequently submitted the liquid-liquid fractionation. The secondary
metabolites were identified by high performance liquid chromatography coupled
with detector diode array (HPLC-DAD) and liquid chromatography diode array
detector coupled with mass spectrometry (LC-DAD-MS). The anti-inflammatory
properties of the aqueous extract, dichloromethane (CH2Cl2), ethyl acetate
(EtOAc) and n-butanol (n-BuOH) fractions of the fruits from H. speciosa, as well
as rutin and chlorogenic acid were investigated using in vitro and in vivo
models. In vivo tests comprised the xylene-induced ear edema that was
measured the formation of edema, carrageenan-induced peritonitis was
evaluated the total leukocytes at 4h and zymosan-induced air pouch was
measured the total leukocytes and differential cell count at 6, 24 and 48 hours,
whereas in vitro tests were evaluated levels of cytokines IL-1?, IL-6, IL-12 and
TNF-? using ELISA obtained of carrageenan-induced peritonitis model. The
results showed the presence of rutin and chlorogenic acid were detected in the
aqueous extract from H. speciosa fruits by HPLC-DAD and LC-DAD-ME.
Furthermore, the aqueous extracts and fractions, as well as rutin and
chlorogenic acid significantly inhibited the xilol-induced ear edema and reduced
cell migration in the animal models such as carrageenan-induced peritonitis and
zymosan-induced air pouch. In addition, reduced levels of cytokines IL-1?, IL-6,
IL-12 and TNF-? were observed. This is the first study that demonstrated the
anti-inflammatory effect of aqueous extract from Hancornia speciosa fruits
against different inflammatory agents in animal models, suggesting that their
bioactive molecules, especially rutin and chlorogenic acid contributing, at least
in part, to the anti-inflammatory effect of aqueous extract. These findings
support the widespread use of Hancornia speciosa in popular medicine and
demonstrate that this aqueous extract has therapeutic potential for the
development of a herbal drugs with anti-inflammatory properties.
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