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

Soyoun Lee; Haeri Wi; Myoungsook Lee

doi:10.4163/kjn.2011.44.6.488

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

Research Article

Korean Journal of Nutrition

Korean Journal of Nutrition 2011; 44(6): 488-497.

Published online: 31 December 2011

DOI: https://doi.org/10.4163/kjn.2011.44.6.488

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

Soyoun Lee, Haeri Wi, Myoungsook Lee

Department of Food and Nutrition and Research Institute of Obesity Sciences, Sungshin Women's University, Seoul 136-742, Korea.

To whom correspondence should be addressed. (mlee@sungshin.ac.kr)

Received 14 November 2011 Revised 29 November 2011 Accepted 13 December 2011

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 µM DAS, 25 µM CAP, 50 µM GGR, and 12.5 µM 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.

Keywords: antioxidant, HepG2 cells, diallyl sulfide, capsaicin, gingerol, sulforaphane

Figures and Tables

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.

Notes

This work was supported by Sungshin Women' University Research grant of 2009.

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