Extracts from the leaves of <i>Campomanesia velutina</i> inhibits production of LPS/INF-γ induced inflammatory mediators in J774A.1 cells and exerts anti-inflammatory and antinociceptive effects <i>in vivo</i>

Michel, Marcela C.P.; Guimarães, Andrea Grabe; Paula, Carmem A.; Rezende, Simone A.; Sobral, Marcos E.G.; Guimarães, Dênia A. Saúde

doi:10.1590/S0102-695X2013000600010

Marcela C.P. Michel

Laboratório de Plantas Medicinais, Escola de Farmácia, Universidade Federal de Ouro Preto (UFOP) Minas Gerais, MG, Brazil

Andrea Grabe Guimarães

Laboratório de Farmacologia Experimental, Escola de Farmácia, Universidade Federal de Ouro Preto (UFOP) Minas Gerais, MG, Brazil

Carmem A. Paula

Laboratório de Plantas Medicinais, Escola de Farmácia, Universidade Federal de Ouro Preto (UFOP) Minas Gerais, MG, Brazil

Simone A. Rezende

Laboratório de Imunoparasitologia, Núcleo de Pesquisas em Ciências Biológicas, Universidade Federal de Ouro Preto, Minas Gerais (UFOP), MG, Brazil

Marcos E.G. Sobral

Departamento de Ciências Naturais, Universidade Federal de São João Del-Rei (UFSJ), Minas Gerais, MG, Brazil

Dênia A. Saúde Guimarães Corresponding author. E-mail: saude@ef.ufop.br(D.A.S. Guimarães).

Laboratório de Plantas Medicinais, Escola de Farmácia, Universidade Federal de Ouro Preto (UFOP) Minas Gerais, MG, Brazil

Corresponding author. E-mail: saude@ef.ufop.br(D.A.S. Guimarães).

Authors' contributions

MCPM (PhD student) contributed in running the laboratory work, analysis of the data and drafted the paper. MEGS contributed in plant identification and herbarium confection. AGG and SAR contributed to biological studies in vivo and in vitro, respectively. CAP contributed to critical reading of the manuscript. DASG designed the study, supervised the laboratory work and contributed to critical reading of the manuscript. All the authors have read the final manuscript and approved the submission.

Figures (5)
Tables (2)

Thumbnail

Figure 1
HPLC chromatograms of ethanolic extract (A), A fraction (B), M fraction (C) and myricitrin (D) obtained by HPLC-UV-DAD at 254 nm.

Thumbnail

Figure 2
A, effect of the ethanolic extract (EE); B, effect of ethyl acetate fraction (A); C, effect of methanolic fraction (M) on NO production by LPS/IFN-γ stimulated J774A.1 macrophages. The cells were treated with EE, A and M dissolved in DMSO at different non-cytotoxic concentrations for 1 h and then stimulated with LPS (25 μg/ml) and IFN-γ (25 UI/ml) for 24 h. Supernatants were collected and nitrite (NO) concentration was determined by Griess reagent. Data are represented as mean ± S.E.M.

*p < 0.05.

**p < 0.01.

***p < 0.001 compared with LPS/IFN-γ-stimulated macrophages alone (ANOVA followed Dunnett’s test).

Thumbnail

Figure 3
A, effect of the ethanolic extract (EE); B, effect of ethyl acetate fraction (A); C, effect of methanolic fraction (M) on TNF-α production by LPS/IFN-γ stimulated J774A.1 macrophages. The cells were treated with EE, A and M dissolved in DMSO at different non-cytotoxic concentrations for 1 h and then stimulated with LPS (25 μg/ml) and IFN-γ (25 UI/ml) for 24 h. Supernatants were collected and TNF-α concentration was determined by specific ELISA kit. Data are represented as mean ± SEM.

*p < 0.05.

**p < 0.01.

***p < 0.001 compared with LPS/IFN-γ-stimulated macrophages alone (ANOVA followed Dunnett’s test).

Thumbnail

Figure 4
A, Effect of the ethanolic extract (EE); B, effect of ethyl acetate fraction (A); C, effect of methanolic fraction (M) on IL-10 production by LPS/IFN-γ stimulated J774.A1 macrophages. The cells were treated with EE, A and M dissolved in DMSO at different non-cytotoxic concentrations for 1 h and then stimulated with LPS (25 μg/ml) and IFN-γ (25 UI/ml) for 24 h. Supernatants were collected and IL-10 concentration was determined by specific ELISA kit. Data are represented as mean ± SEM. *p < 0.05. **p < 0.01. ***p < 0.001 compared with LPS/IFN-γ-stimulated macrophages alone (ANOVA followed Dunnett’s test).

Thumbnail

Figure 5
Effect of myricitrin on NO (A), TNF-α (B) and IL-10 (C) production by LPS/IFN-γ stimulated J774.A1 macrophages. The cells were treated with myricitrin dissolved in DMSO at different non-cytotoxic concentrations for 1 h and then stimulated with LPS (25 μg/ml) and IFN-γ (25 UI/ml) for 24 h. Supernatants were collected and nitrite (NO) concentration was determined by Griess reagent, TNF-α and IL-10 concentration was determined by specific ELISA kit. Data are represented as mean ± S.E.M.

*p < 0.05.

**p < 0.01.

***p < 0.001 compared with LPS/IFN-γ-stimulated macrophages alone (ANOVA followed Dunnett’s test).

Thumbnail

Table 1
Effect of oral administration of ethanolic Campomanesia velutina extract (EE) and its ethyl acetate (A) and methanolic (M) fractions on carrageenan-induced paw edema.

Thumbnail

Table 2
Effect of oral administration of ethanolic Campomanesia velutina extract (EE) and its ethyl acetate (A) and methanolic (M) fractions in the acetic acid-induced abdominal writhing test.

Treatment	Swelling thickness (%)
(After carrageenan)	1 h	2 h	3 h	4 h	5 h	6 h
Control condition^a a Without carrageenan.	1,88 ± 1,10^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	0,27 ± 3,32^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	4,14 ± 2,81^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	4,27 ± 3,12^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	3,01 ± 1,95^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	1,05 ± 1,56^c c p < 0.01.
Vehicle	14,22 ± 1,84	22,00 ± 2,84	30,22 ± 3,10	25,58 ± 3,99	18,36 ± 4,19	12,32 ± 2,97
Indomethacin	6,62 ± 1,93	11,04 ± 1,93^b b p < 0.05.	15,25 ± 2,79^c c p < 0.01.	10,65 ± 2,25^c c p < 0.01.	5,80 ± 2,61^c c p < 0.01.	0,72 ± 2,18^c c p < 0.01.
EE 100 mg/kg	13,77 ± 2,89	17,97 ± 3,10	18,26 ± 3,64^b b p < 0.05.	13,24 ± 3,87^b b p < 0.05.	8,10 ± 2,59^b b p < 0.05.	1,51 ± 1,90^c c p < 0.01.
EE 300 mg/kg	4,86 ± 2,60^b b p < 0.05.	11,47 ± 2,67^b b p < 0.05.	14,40 ± 2,98^c c p < 0.01.	11,20 ± 2,93^c c p < 0.01.	6,92 ± 1,86^b b p < 0.05.	1,53 ± 1,38^c c p < 0.01.
A 100 mg/kg	12,24± 2,09	15,74 ± 2,57	10,50 ± 3,62^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	15,43± 3,69	22,09 ± 2,66	2,00 ± 2,33^c c p < 0.01.
A 300 mg/kg	12,86 ± 1,64	15,01 ± 1,69	4,70 ± 2,39^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	15,14 ± 1,59	18,70 ± 2,33	2,55 ± 2,49^c c p < 0.01.
M 100 mg/kg	11,96 ± 2,01	13,67 ± 1,61	19,27 ± 2,26	11,67 ± 1,60^b b p < 0.05.	5,85 ± 1,65^b b p < 0.05.	2,36 ± 1,37^c c p < 0.01.
M 300 mg/kg	11,78 ± 1,89	13,15 ± 1,30	7,99 ± 1,86^d d p < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).	8,20 ± 3,17'	7,41 ± 1,32^b b p < 0.05.	2,54 ± 1,32^c c p < 0.01.

Treatment	Number of writhes	Inhibition (%)
Vehicle	58.62 ± 7.09	-
Indomethacin	18.37 ± 5.44	68.66^b bp < 0.01.
EE 100 mg/kg	34.37 ± 7.53	41.37
EE 300 mg/kg	9.37 ± 4.59	84.01^c cp < 0.001, when compared to the group treated with vehicle (ANOVA followed Dunnett's test).
A 100 mg/kg	20.25 ± 7.09	65.45^a ap < 0.05
A 300 mg/kg	16.75 ± 8.19	71.43
M 100 mg/kg	33.00 ± 13.93	43.71
M 300 mg/kg	38.71 ± 10.08	33.96

[1] Corresponding author. E-mail: saude@ef.ufop.br(D.A.S. Guimarães).

Brasil

Brasil

Extracts from the leaves of Campomanesia velutina inhibits production of LPS/INF-γ induced inflammatory mediators in J774A.1 cells and exerts anti-inflammatory and antinociceptive effects in vivo