Nutr Res Pract.  2014 Apr;8(2):138-145.

Antioxidative effects of fermented sesame sauce against hydrogen peroxide-induced oxidative damage in LLC-PK1 porcine renal tubule cells

Affiliations
  • 1Department of Food Science and Nutrition, Pusan National University, Busan 609-735, Korea. kunypark@pusan.ac.kr
  • 2Kimchi Research Institute, Pusan National University, 2, Busandaehak-ro 63 Beon-gil, Geumjeong, Busan 609-735, Korea.
  • 3Daesang R&D, Icheon, Gyeonggi 467-813, Korea.

Abstract

BACKGROUND/OBJECTIVES
This study was performed to investigate the in vitro antioxidant and cytoprotective effects of fermented sesame sauce (FSeS) against hydrogen peroxide (H2O2)-induced oxidative damage in renal proximal tubule LLC-PK1 cells.
MATERIALS/METHODS
1,1-diphenyl-2-picrylhydrazyl (DPPH), hydroxyl radical (*OH), and H2O2 scavenging assay was used to evaluate the in vitro antioxidant activity of FSeS. To investigate the cytoprotective effect of FSeS against H2O2-induced oxidative damage in LLC-PK1 cells, the cellular levels of reactive oxygen species (ROS), lipid peroxidation, and endogenous antioxidant enzymes including catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GSH-px) were measured.
RESULTS
The ability of FSeS to scavenge DPPH, *OH and H2O2 was greater than that of FSS and AHSS. FSeS also significantly inhibited H2O2-induced (500 microM) oxidative damage in the LLC-PK1 cells compared to FSS and AHSS (P < 0.05). Following treatment with 100 microg/mL of FSeS and FSS to prevent H2O2-induced oxidation, cell viability increased from 56.7% (control) to 83.7% and 75.6%, respectively. However, AHSS was not able to reduce H2O2-induced cell damage (viability of the AHSS-treated cells was 54.6%). FSeS more effectively suppressed H2O2-induced ROS generation and lipid peroxidation compared to FSS and AHSS (P < 0.05). Compared to the other sauces, FSeS also significantly increased cellular CAT, SOD, and GSH-px activities and mRNA expression (P < 0.05). CONCULUSIONS: These results from the present study suggest that FSeS is an effective radical scavenger and protects against H2O2-induced oxidative damage in LLC-PK1 cells by reducing ROS levels, inhibiting lipid peroxidation, and stimulating antioxidant enzyme activity.

Keyword

Oxidative stress; fermented sesame sauce; antioxidant enzymes; LLC-PK1 cells

MeSH Terms

Animals
Catalase
Cats
Cell Survival
Glutathione Peroxidase
Hydrogen Peroxide
Hydrogen*
Hydroxyl Radical
Lipid Peroxidation
LLC-PK1 Cells
Oxidative Stress
Reactive Oxygen Species
RNA, Messenger
Sesamum*
Superoxide Dismutase
Swine
Catalase
Glutathione Peroxidase
Hydrogen
Hydrogen Peroxide
Hydroxyl Radical
RNA, Messenger
Reactive Oxygen Species
Superoxide Dismutase

Figure

  • Fig. 1 DPPH (A) and •OH radical scavenging (B) activities of the sauce extracts. Data from three independent experiments are presented as the mean ± SD. a-fDifferent letters over the bars indicate that the mean values are significantly different (P < 0.05) according to Duncan's multiple range test. AHSS, acid-hydrolyzed soy sauce extract; FSS, fermented soy sauce extract; FSeS, fermented sesame sauce extract.

  • Fig. 2 H2O2 scavenging activities of the sauce extracts. Data from three independent experiments are presented as the mean ± SD. a-dDifferent letters over the bars indicate that the mean values are significantly different (P < 0.05) according to Duncan's multiple range test. AHSS, acid-hydrolyzed soy sauce extract; FSS, fermented soy sauce extract; FSeS, fermented sesame sauce extract.

  • Fig. 3 Protective effect of the sauce extracts against oxidative stress induced by H2O2 in LLC-PK1 cells. Data from three independent experiments are presented as the mean ± SD. a-fDifferent letters over the bars indicate that the mean values are significantly different (P < 0.05) according to Duncan's multiple range test. AHSS, acid-hydrolyzed soy sauce extract; FSS, fermented soy sauce extract; FSeS, fermented sesame sauce extract.

  • Fig. 4 Effect of sauce extracts on ROS generation in LLC-PK1 cells treated with H2O2. Data from three independent experiments are presented as the mean ± SD. a-eDifferent letters over the bars indicate that the mean values are significantly different (P < 0.05) according to Duncan's multiple range test. AHSS, acid-hydrolyzed soy sauce extract; FSS, fermented soy sauce extract; FSeS, fermented sesame sauce extract.

  • Fig. 5 Effect of sauce extracts on lipid peroxidation levels in LLC-PK1 cells treated with H2O2. Data from three independent experiments are presented as the mean ± SD. a-fDifferent letters over the bars indicate that the mean values are significantly different (P < 0.05) according to Duncan's multiple range test. AHSS, acid-hydrolyzed soy sauce extract; FSS, fermented soy sauce extract; FSeS, fermented sesame sauce extract.

  • Fig. 6 Effects of the sauce extracts (100 µg/mL) on mRNA expression of CAT, SOD, GSH-px, and GST in LLC-PK1 cells treated with H2O2(500 µM). Band intensity was measured with a densitometer and expressed as fold of the control. Fold ratio = gene expression/β-actin × control numerical value (control fold ratio: 1). a-dDifferent letters over the bars indicate that the mean values are significantly different (P < 0.05) according to Duncan's multiple range test. AHSS, acid-hydrolyzed soy sauce extract; FSS, fermented soy sauce extract; FSeS, fermented sesamin sauce extract.


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