Korean J Nutr.  2011 Dec;44(6):488-497. 10.4163/kjn.2011.44.6.488.

Comparison of the Antioxidant Effects of Diallyl Sulfide, Capsaicin, Gingerol and Sulforaphane in H2O2-Stressed HepG2 Cells

• Lee S
• Wi H
• Lee M

• KMID: 2268609
• DOI: http://doi.org/10.4163/kjn.2011.44.6.488

Abstract

Oxygen is necessary to sustain life, yet cellular oxygen metabolism creates destructive elements called free radicals. Free radicals are chemically unbalanced and carrying free electrons that can damage molecules, potentially damaging the cell itself. For this reason, many antioxidant products, including supplements and functional foods, are being developed. In particular, natural products are rich sources of pharmacologically active compounds. The purpose of this study was to investigate the antioxidant effects of target biomaterials in Korean traditional spices such as diallyl sulfide (DAS), capsaicin (CAP), and gingerol (GGR), and to investigate the response of the antioxidant defense system to oxidative stress by hydrogen peroxide (H2O2) compared to sulforaphane (SFN) in HepG2 cells. After the analysis of the cell viability using Cell Counting kit-8 (CCK-8) assay, we determined that the optimum levels were 200 microM DAS, 25 microM CAP, 50 microM GGR, and 12.5 microM SFN. Antioxidant enzymes were measured and protein expression was detected by Western blotting. All treatments showed a significant decrease in antioxidant enzyme activity such as superoxide dismutase, catalse, and glutathione peroxidase in HepG2 cells. Additionally, DAS, CAP, GGR and SFN increased the antioxidant system-related transcription factor Nrf2 which was found to be regulated by the activation of MAPK-JNK in this study. In conclusion, these results indicate the protective effects of DAS CAP, GGR, and SFN against H2O2-induced oxidative stress.

Keyword

antioxidant; HepG2 cells; diallyl sulfide; capsaicin; gingerol; sulforaphane

MeSH Terms

4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid
Allyl Compounds
Antioxidants
Biocompatible Materials
Biological Agents
Blotting, Western
Capsaicin
Catechols
Cell Count
Cell Survival
Electrons
Fatty Alcohols
Free Radicals
Functional Food
Glutathione Peroxidase
Hep G2 Cells
Hydrogen Peroxide
Lifting
Oxidative Stress
Oxygen
Spices
Sulfides
Superoxide Dismutase
Thiocyanates
Transcription Factors
4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid
Allyl Compounds
Antioxidants
Biocompatible Materials
Biological Agents
Capsaicin
Catechols
Fatty Alcohols
Free Radicals
Glutathione Peroxidase
Hydrogen Peroxide
Oxygen
Sulfides
Superoxide Dismutase
Thiocyanates
Transcription Factors

Figure

• 

  Fig. 1 Cell viability of DAS, CAP, GGR and SFN in H2O2 induced Human Hepatoma HepG2 Cells. DAS: diallyl sulfide, CAP: capsaicin, GGR: gingerol, SFN: sulforaphane.

• 

  Fig. 2 Antioxidant effects of DAS, CAP, GGR and SFN on the activities of SOD (A) and CAT (B) in H2O2 induced Human Hepatoma HepG2 Cells. DAS: diallyl sulfide, CAP: capsaicin, GGR: gingerol, SFN:sulforaphane. Data are presented as mean ± SD. a, b, c: Significant difference between the groups were tested by ANOVA. *: p < 0.05, **: p < 0.01, ***: p < 0.001: Significantly different each groups compared to negative control (only H2O2).

• 

  Fig. 3 Effects of DAS, CAP, GGR and SFN on the levels of GSH (A) and GPx activity (B) in H2O2 induced Human Hepatoma HepG2 Cells. DAS: diallyl sulfide, CAP: capsaicin, GGR: gingerol, SFN: sulforaphan. Data are presented as mean ± SD. a, b, c: Significant difference between the groups were tested by ANOVA. *: p < 0.05, **: p < 0.01, ***: p < 0.001: Significantly different each groups compared to negative control (only H2O2).

• 

  Fig. 4 Effect of DAS, CAP, GGR and SFN on LPO reduction in H2O2 induced Human Hepatoma HepG2 Cells. DAS: diallyl sulfide, CAP: capsaicin, GGR: gingerol, SFN: sulforaphan. Data are presented as mean ± SD. a, b, c : Significant difference between the groups were tested by ANOVA. *: p < 0.05, **: p < 0.01, ***: p < 0.001: Significantly different each groups compared to negative control (only H2O2).

• 

  Fig. 5 Effects of DAS, CAP, GGR and SFN on the expression of oxidative stress markers, pIkBα (A), NFkB (B), TNFα (C) against H2O2-pretreated HepG2 cells. Actin levels were compared to ensure equal amount of protein loading. DAS: diallyl sulfide, CAP: capsaicin, GGR: gingerol, SFN: sulforaphan.

• 

  Fig. 6 Effects of DAS, CAP, GGR and SFN on the expression of antioxidative signals, JNK and pJNK (A), Nrf2 (B), pNrf2 (C) against H2O2-pretreated HepG2 cells. Actin levels were compared to ensure equal amount of protein loading. DAS: diallyl sulfide, CAP: capsaicin, GGR: gingerol, SFN: sulforaphan.

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