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. 2014 Jun;19(2):89-97.
doi: 10.3746/pnf.2014.19.2.089.

Antioxidant and Anti-inflammatory Activities of Broccoli Florets in LPS-stimulated RAW 264.7 Cells

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Free PMC article

Antioxidant and Anti-inflammatory Activities of Broccoli Florets in LPS-stimulated RAW 264.7 Cells

Joon-Ho Hwang et al. Prev Nutr Food Sci. .
Free PMC article

Abstract

Broccoli (Brassica oleracea var. italia) florets were extracted with 80% methanol and the extract was sequentially fractionated with n-hexane, ethyl acetate, n-butanol, and distilled water. The extract and the fractions were evaluated for total phenolic content, sulforaphane content, antioxidant activity, and anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. The total phenolic content and sulforaphane content of the ethyl acetate fraction (EF) were 35.5 mg gallic acid equivalents/g and 620.2 μg/g, respectively. These values were higher than those of the 80% methanol extract and organic solvent fractions. The oxygen radical absorbance capacity of the EF [1,588.7 μM Trolox equivalents (TE)/mg] was 11-fold higher than that of the distilled water fraction (143.7 μM TE/mg). The EF inhibited nitric oxide release from LPS-stimulated RAW 264.7 cells in a dose-dependent manner and inhibited IκB-α degradation and nuclear factor-κB activation in LPS-stimulated RAW 264.7 cells. In conclusion, the EF of broccoli florets exerted potent antioxidant and anti-inflammatory effects.

Keywords: anti-inflammatory activity; antioxidant activity; broccoli florets; sulforaphane; total phenolics.

Figures

Fig. 1
Fig. 1
Cell viability (□) and inhibitory activity of nitric oxide production (-•-) by LPS (100 ng/mL)-stimulated RAW 264.7 cells treated with an 80% methanol extract and organic solvent fractions of broccoli florets. (A) 80% methanol extract, (B) n-hexane fraction, (C) ethyl acetate fraction, (D) n-butanol fraction, (E) distilled water fraction. Values with different letters (A–F, a–g) are significantly different at P<0.05 according to Duncan’s multiple range test.
Fig. 1
Fig. 1
Cell viability (□) and inhibitory activity of nitric oxide production (-•-) by LPS (100 ng/mL)-stimulated RAW 264.7 cells treated with an 80% methanol extract and organic solvent fractions of broccoli florets. (A) 80% methanol extract, (B) n-hexane fraction, (C) ethyl acetate fraction, (D) n-butanol fraction, (E) distilled water fraction. Values with different letters (A–F, a–g) are significantly different at P<0.05 according to Duncan’s multiple range test.
Fig. 2
Fig. 2
Effects of the ethyl acetate fraction of broccoli florets on iNOS, TNF-α, IL-1β, and IL-6 mRNA levels in LPS (100 ng/mL)-induced RAW 264.7 cells. (A) RT-PCR analysis of mRNA transcripts, (B) relative levels of mRNA expression (normalized to β-actin). Values with different letters (a–f) are significantly different at P<0.05 according to Duncan’s multiple range test.
Fig. 3
Fig. 3
Effects of the ethyl acetate fraction of broccoli florets on iNOS protein levels in LPS (100 ng/mL)-induced RAW 264.7 cells. (A) Immunoblotting analysis of iNOS protein, (B) relative levels of iNOS protein (normalized to β-actin). Values with different letters (a–e) are significantly different at P<0.05 according to Duncan’s multiple range test.
Fig. 4
Fig. 4
Effects of the ethyl acetate fraction of broccoli florets on the inhibition of (A) NF-κB-luciferase activity and (B) I-κBα degradation in LPS (100 ng/mL)-induced in RAW 264.7 cells (Western blot analysis). Values with different letters (a–f) are significantly different at P<0.05 according to Duncan’s multiple range test.

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