Nutr Res Pract.  2010 Feb;4(1):3-10.

Desalinated underground seawater of Jeju Island (Korea) improves lipid metabolism in mice fed diets containing high fat and increases antioxidant potential in t-BHP treated HepG2 cells

• Noh JR
• Gang GT
• Kim YH
• Yang KJ
• Lee CH
• Na OS
• Kim GJ
• Oh WK
• Lee YD

Abstract

This study was performed to investigate the effect of desalinated underground seawater (named as 'magma seawater', MSW) of Jeju Island in Korea on lipid metabolism and antioxidant activity. MSW was collected from underground of Han-Dong in Jeju Island, and freely given to high fat diet (HFD)-fed C57BL/6 mice for 10 weeks. Although there were no significant differences in the body weight changes and plasma lipid levels, hepatic triglyceride levels were significantly lower in the MSW group than in the normal tap water (TW)-drunken control group. Furthermore, the activity of fatty acid synthase (FAS) was significantly decreased and carnitine palmitoyltransferase (CPT) activity was increased in MSW group compared to TW group. Similarly, real-time PCR analysis revealed that mRNA expressions of lipogenic genes were lowered in MSW groups compared to the control group. In a morphometric observation on the liver tissue, accumulation of fats was remarkably reduced in MSW group. Meanwhile, in vitro assay, free radical scavenging activity measured by using diphenylpicrylhydrazyl (DPPH) was increased in MSW group. The 2'-7'-dichlorofluorescein diacetate (DCF-DA) staining followed with fluorescent microscopy showed a low intensity of fluorescence in MSW-treated HepG2 cells, compared to TW-treated HepG2 cells, which indicated that the production of reactive oxygen species by tert-butyl hydroperoxide (t-BHP) in HepG2 cells was decreased by MSW treatment. The antioxidant effect of MSW on t-BHP-induced oxidative stress in HepG2 cells was supported by the increased activities of intracellular antioxidant enzymes such as catalase and glutathione reductase. From these results, we speculate that MSW has an inhibitory effect on lipogenesis in liver and might play a protective role against cell damage by t-BHP-induced oxidative stress.

Keyword

Underground seawater; lipid metabolism; hepatic lipid; antioxidant effect

MeSH Terms

Animals
Antioxidants
Body Weight Changes
Carnitine O-Palmitoyltransferase
Catalase
Diet
Diet, High-Fat
Fats
Fatty Acid Synthetase Complex
Fluorescence
Glutathione Reductase
Hep G2 Cells
Korea
Lipid Metabolism
Lipogenesis
Liver
Mice
Microscopy
Oxidative Stress
Plasma
Reactive Oxygen Species
Real-Time Polymerase Chain Reaction
RNA, Messenger
Seawater
tert-Butylhydroperoxide
Water
Antioxidants
Carnitine O-Palmitoyltransferase
Catalase
Fats
Fatty Acid Synthetase Complex
Glutathione Reductase
RNA, Messenger
Reactive Oxygen Species
Water
tert-Butylhydroperoxide

Figure

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  Fig. 1 Weekly changes of body weight and water intake in C57BL/6 mice treated with high fat diet (HFD) and drinking of desalinated underground seawater. Values are mean ± SEM (n=10, each group).

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  Fig. 2 Plasma and hepatic lipid concentrations determined in C57BL/6 mice treated with high fat diet (HFD) and drinking of desalinated underground seawater. Value are mean ± SEM. *Significantly different from tap water control group at P < 0.05 (student's t-test). MSW: magma seawater (desalinated underground seawater), TW: tap water.

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  Fig. 3 Representative photomicrographs showing inhibited fatty liver in C57BL/6 mice treated with high fat diet (HFD) and drinking of desalinated underground seawater. Values are mean ± SEM (n=5, each group). MSW: magma seawater (desalinated underground seawater), TW: tap water.

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  Fig. 4 Quantitative mRNA expressions of liver lipogenesis genes in C57BL/6 mice treated with high fat diet (HFD) and drinking of desalinated underground seawater. Values are mean ± SEM. The normalization of the gene was carried out 18s RNA expression for each group. FAS: fatty acid synthase, HMG-CR: -hydroxy-3-methyl-glutaryl-CoA reductase, ACC: acetyl-CoA carboxylase, SREBP1c: sterol regulatory element binding protein 1c. MSW: magma seawater (desalinated underground seawater), TW: tap water.

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  Fig. 5 Activities of hepatic lipid regulating enzymes in C57BL/6 mice treated with high fat diet (HFD) and drinking of desalinated underground seawater. Values are mean ± SEM. *Significantly different from tap water control group at P < 0.05 (student's t-test). FAS: fatty acid synthase, CPT: carnitine palmitoyltransferase. MSW: magma seawater (desalinated underground seawater), TW: tap water.

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  Fig. 6 Effect of desalinated underground seawater on DPPH radical scavenging activity. The relative DPPH scavenging activities of desalinated underground seawater to the tap water group are expressed as percentage values (mean ± SEM.) of triplicated determinations, and bars with different letters indicate significant differences at P < 0.05 (one-way ANOVA) among groups. Fraction numbers represent two-fold serial dilutions of desalinated underground seawater. MSW: magma seawater (desalinated underground seawater), TW: tap water.

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  Fig. 7 Represntative fluorescence microscopic findings on ROS production induced by t-BHP in desalinated underground seawater-treated HepG2 cells. Reduced fluorescence intensity was observed in desalinated underground seawater group. MSW: magma seawater (desalinated underground seawater), TW: tap water. DCF-DA (2'-7'-dichlorofluorescein diacetate) staining, magnification × 200.

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  Fig. 8 Effect of desalinated underground seawater effect on antioxidant enzyme activities in t-BHP-treated HepG2 cells. Intracellular antioxidant enzyme activities were represented as mean ± SEM. of triplicated determination. *Significantly different from tap water control group at P < 0.05 (student's t-test). CAT: catalase, SOD: superoxide dismutase, GPx: glutathione peroxidase, GR: glutathione reductase, MSW: magma seawater (desalinated underground seawater), TW: tap water.

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