Nutr Res Pract.  2019 Jun;13(3):189-195. 10.4162/nrp.2019.13.3.189.

Anti-inflammatory effect of aged black garlic on 12-O-tetradecanoylphorbol-13-acetate-induced dermatitis in mice

• You BR
• Yoo JM
• Baek SY
• Kim MR

• KMID: 2453282
• DOI: http://doi.org/10.4162/nrp.2019.13.3.189

Abstract

BACKGROUND/OBJECTIVES
Although aged black garlic has various biological activities such as anti-allergy, anti-inflammation and neuroprotection, effect of aged black garlic on chemically contact dermatitis is unclarified.
MATERIALS/METHODS
To evaluate anti-dermatitic activity of aged black garlic extract, we investigated effects of a fraction of aged black garlic extract (BG10) on both in vivo and in vitro.
RESULTS
BG10 almost inhibited formation of nitric monoxide and interleukin-6 (IL-6; IC50, 7.07 µg/mL) at 25 µg/mL, and dose-dependently reduced production of tumor necrosis factor-α (TNF-α; IC50, 52.07 µg/mL) and prostaglandin E2 (IC50, 38.46 µg/mL) in lipopolysaccharide-stimulated RAW264.7 cells. In addition, BG10 significantly inhibited the expression of inducible nitric oxide synthase, cyclooxygenase-2 and nuclear NF-κB, and improved that of cytosolic levels of NF-κB and IκBα in the cells. Consistent with in vitro studies, BG10 (0.5 mg/mL) not only reduced ear edema but also suppressed the formation of IL-6 and TNF-α induced by 12-O-tetradecanoylphorbol-13-acetate in ear tissues of mice.
CONCLUSIONS
These findings suggest BG10 has anti-dermatitic activity through inhibiting activation of macrophages. Therefore, such effects of BG10 may provide information for the application of aged black garlic for prevention and therapy of contact dermatitis.

Keyword

Contact dermatitis; COX-2; cytokines; macrophage; NF-kappa B

MeSH Terms

Animals
Cyclooxygenase 2
Cytokines
Cytosol
Dermatitis*
Dermatitis, Contact
Dinoprostone
Ear
Edema
Garlic*
In Vitro Techniques
Inhibitory Concentration 50
Interleukin-6
Macrophages
Mice*
Necrosis
Neuroprotection
NF-kappa B
Nitric Oxide Synthase Type II
Cyclooxygenase 2
Cytokines
Dinoprostone
Interleukin-6
NF-kappa B
Nitric Oxide Synthase Type II

Figure

• 

  Fig. 1 Inhibitory effects of BG10 on TPA-induced dermatitis in mice. ICR mice were spread with BG10, a fraction of aged black garlic extract (0.5 mg/mL), on the back of ears prior to 12-O-tetradecanoylphorbol-13-acetate (TPA; 0.1 mg/mL) challenge. Ear thickness, and the levels of IL-6 and TNF-α in ear tissues were determined as described in the Materials and Methods section. The data are expressed as the mean ± SEM values of septuple determinations. **P < 0.01 versus the TPA-treated group. A, body weight; B, food intake; C, ear thickness; D, IL-6; E, TNF-α.

• 

  Fig. 2 Inhibitory effects of BG10 on the formation of NO, IL-6, TNF-α and PGE2 in LPS-activated RAW 264.7 cells. RAW 264.7 cells were seeded on 96-well plate (5 × 104 cells/well) in DMEM with 10% FBS at 37℃ overnight. The cells were simultaneously exposed to BG10 (0–100 µg/mL) with or without lipopolysaccharide (LPS; 1 µg/mL) for 24 h. The levels of NO, IL-6, TNF-α and PGE2 were determined as described in the Materials and Methods section. The data are expressed as the mean ± SD values of triple determinations. **P < 0.01 versus the LPS-treated group. A, NO; B, IL-6; C, TNF-α; D, PGE2.

• 

  Fig. 3 Effect of BG10 on cell viability in LPS-activated RAW 264.7 cells. RAW 264.7 cells were seeded on 96-well plate in DMEM with 10% FBS at 37℃ overnight. The cells were simultaneously exposed to BG10 with or without LPS for 22 h, and then further incubated with methyl thiazolyl tetrazolium (MTT) reagent (500 µg/mL) for 2 h. Cell viability was determined as described in Materials and Methods section. The data are expressed as the mean ± SD values of quadruple determinations.

• 

  Fig. 4 Effects of of BG10 on the expression of iNOS, COX-2, nuclear NF-κB, or cytosolic NF-κB and IκB. RAW 264.7 cells were seeded on 6-well plate (1 × 106 cells/well) in DMEM with 10% FBS at 37℃ overnight. The cells were simultaneously exposed to BG10 with or without LPS for 15 min or 6 h. They were washed, and then lysed in a cell lysis buffer. The expression of iNOS, COX-2 and nuclear NF-κB as well as cytosolic NF-κB and IκBα was determined as described in Materials and Methods section. Similar results were obtained in three independent experiments. **P < 0.01 versus LPS-treated group. A, iNOS and COX-2; B, nuclear NF-κB, cytosolic NF-κB and cytosolic IκBα.

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