Korean J Physiol Pharmacol.  2014 Feb;18(1):33-39. 10.4196/kjpp.2014.18.1.33.

Anti-Inflammatory and Anti-Superbacterial Properties of Sulforaphane from Shepherd's Purse

• Choi WJ
• Kim SK
• Park HK
• Sohn UD
• Kim W

• KMID: 2285488
• DOI: http://doi.org/10.4196/kjpp.2014.18.1.33

Abstract

Shepherd's purse, Capsella bursa-pastoris (L.) Medik., has been considered a health food for centuries in Asia and is known to contain the isothiocyanate compound sulforaphane. In this study, we evaluated the anti-inflammatory and antibacterial properties of a sulforaphane-containing solution (SCS) isolated from shepherd's purse. SCS had significant anti-inflammatory activity indicated by the decreased levels of nitric oxide (NO), cytokines (interleukin 1beta [IL-1beta], IL-6, and IL-10), and prostaglandin E2 (PGE2) in lipopolysaccharide-stimulated RAW 264.7 murine macrophages. In addition, SCS decreased the inducible NO synthase (iNOS) and cyclooxygenase 2 (COX-2) levels, which confirmed the anti-inflammatory activity of SCS. Further, SCS inhibited vancomycin-resistant enterococci (VRE) and Bacillus anthracis. The minimal inhibitory concentration was 250 microg/ml for VRE and 1,000 microg/ml for B. anthracis. Taken together, these data indicate that SCS has potential anti-inflammatory and anti-superbacterial properties, and thus it can be used as a functional food or pharmaceutical.

Keyword

Anti-inflammatory effect; Antimicrobial activity; Cytokine; RAW 264.7 murine macrophages; Shepherd's purse; Sulforaphane

MeSH Terms

Asia
Bacillus anthracis
Capsella*
Cyclooxygenase 2
Cytokines
Dinoprostone
Functional Food
Food, Organic
Interleukin-6
Macrophages
Nitric Oxide
Nitric Oxide Synthase
Cyclooxygenase 2
Cytokines
Dinoprostone
Interleukin-6
Nitric Oxide
Nitric Oxide Synthase

Figure

• 

  Fig. 1 The cell cytotoxicity of RAW 264.7 cells treated with different concentrations of sulforaphane-containing solution (SCS). Cells were treated with 0, 1, 5, 10, 50, 100, or 500 µg/ml SCS in the presence of 0.1 µg/ml lipopolysaccharide (LPS) for 24 h. The normal group was treated with media only. Results are expressed as mean±standard error (SE) from 3 independent experiments. *p<0.01 indicates significant differences from the LPS treated cells.

• 

  Fig. 2 Suppression of lipopolysaccharide (LPS)-induced nitric oxide (NO) and prostaglandin E2 (PGE2) production in RAW 264.7 cells using sulforaphane-containing solution (SCS). Cells were treated with 0, 1, 5, 10, 50, 100, or 500 µg/ml SCS in the presence of 0.1 µg/ml LPS for 18 h. The normal group was treated with media only. Results are expressed as mean±standard error (SE) from 3 independent experiments. *p<0.01 indicates significant differences from the LPS treated cells.

• 

  Fig. 3 Suppression of lipopolysaccharide (LPS)-induced cytokine production (interleukin 1β [IL-1β], IL-6, and IL-10 in RAW 264.7 cells using sulforaphane-containing solution (SCS). RAW 264.7 cells were treated with 0, 1, 5, 10, 50, 100, or 500 µg/ml SCS in the presence of 0.1 LPS for 18 h. The normal group was treated with media only. Results are expressed as mean±standard error (SE) from 3 independent experiments. *p<0.01 indicates significant differences from the LPS treated cells.

• 

  Fig. 4 Effects of sulforaphane-containing solution (SCS) on lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) protein expression. Inhibitory effect of SCS on LPS-induced iNOS and COX-2 protein expression in RAW 264.7 cells. Cells were treated with 0, 1, 5, 10, 50, 100, or 500 µg/ml SCS for 24 h. Cell lysates were electrophoretically separated and transferred, and expression levels of iNOS and COX-2 were detected with specific antibodies. β-actin was used as an internal control for western blot analysis.

• 

  Fig. 5 Effects of sulforaphane-containing solution (SCS) on inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) mRNA levels in lipopolysaccharide (LPS)-stimulated RAW 264.7 cells. Cells were treated with LPS or LPS with SCS at concentrations indicated in Fig. 5. After 24 h of incubation, total RNA was subjected to qRT-PCR. The RT products were labeled with SYBR Green dye. Relative iNOS and COX-2 mRNA expression (2-ΔΔCt) was determined by real-time PCR and calculated by Ct values for iNOS and COX-2 from β-actin mRNA. ΔΔCt=(Cttarget gene-Ctβ-actin)-(Ctcontrol-Ctβ-actin). Each value represents mean±SE of 3 independent experiments. *p<0.01 indicates significant differences from the LPS treated cells.

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