Nutr Res Pract.  2013 Jun;7(3):153-159.

Oxidation of fatty acid may be enhanced by a combination of pomegranate fruit phytochemicals and acetic acid in HepG2 cells

• Kim JY
• Ok E
• Kim YJ
• Choi KS
• Kwon O

Abstract

We investigated whether the combination of phytochemicals and acetic acid in the form of fruit vinegar provides an additive effect on changes of mRNA levels related to fatty acid oxidation in human hepatocyte (HepG2). Among the seven fruit vinegars (Rubuscoreanus, Opuntia, blueberry, cherry, red ginseng, mulberry, and pomegranate) studied, treatment of HepG2 with pomegranate vinegar (PV) at concentrations containing 1 mM acetic acid showed the highest in vitro potentiating effect on the mRNA expression levels of peroxisome proliferator-activated receptor alpha, carnitinepalmitoyl transferase-1, and acyl-CoA oxidase compared to the control group (P < 0.05). Reversed-phase liquid chromatography in combination with quadrupole time-of-flight mass spectrometry analysis revealed four potential compounds (punicalagin B, ellagic acid, and two unidentified compounds) responsible for altered gene expression in HepG2 cells treated with PV as compared with the others. Further investigations are warranted to determine if drinking PV beverages may help to maintain a healthy body weight in overweight subjects.

Keyword

Vinegar beverage; acetic acid; pomegranate; fatty acid oxidation; HepG2 cells

MeSH Terms

Acetic Acid
Acyl-CoA Oxidase
Beverages
Blueberry Plant
Body Weight
Chromatography, Reverse-Phase
Drinking
Ellagic Acid
Fruit
Gene Expression
Hep G2 Cells
Hepatocytes
Humans
Mass Spectrometry
Morus
Opuntia
Overweight
Panax
PPAR alpha
Prunus
Punicaceae
RNA, Messenger
Acetic Acid
Acyl-CoA Oxidase
Ellagic Acid
PPAR alpha
RNA, Messenger

Figure

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  Fig. 1 Comparison of seven fruit vinegars on the mRNA levels of PPARα (A), CPT-1 (B), ACO (C), and UCP-2 (D) in HepG2 cells. Each sample containing 1 mM acetic acid was applied to HepG2 cells, and the cells were incubated for 3 h. The mRNA levels were measured by RT-PCR and normalized to β-actin. Data shown (mean ± SD, n = 3) are typical of three experiments with similar results. *P < 0.05 vs. control, based on ANOVA with the post-hoc Dunnett's multiple range test.

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  Fig. 2 Combination effect of PV on mRNA levels of PPARα, CPT-1, and ACO in HepG2 cells. PV containing 1 mM acetic acid or 1 mM acetic acid alone was applied to HepG2 cells, and the cells were incubated for 3 h. The mRNA levels were measured by RT-PCR and normalized to β-actin. Data shown (mean ± SD, n = 3) are typical of three experiments with similar results. *P < 0.05 vs. control, based on ANOVA with the post-hoc Dunnett's multiple range test.

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  Fig. 3 Dose-dependency of PV on the mRNA levels of PPARα (◆), CPT-1 (□), and ACO (●) in HepG2 cells. PV containing 0.25 to 1 mM acetic acid was applied to HepG2 cells, and the cells were incubated for 3 h. The mRNA levels were measured by RT-PCR and normalized to β-actin. Data shown (mean ± SD, n = 3) are typical of three experiments with similar results.

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  Fig. 4 Comparison of phytochemical profiling of seven fruit vinegars. (A) shows overlaid chromatogram of seven fruit vinegars. HPLC chromatogram magnifications of panel 1 (B), panel 2 (C), and panel 3 (D) shows the four components detected in the PV only. The relative retention time and the area of PV-specific peaks (down arrow) are indicated in the chromatogram. Panel 1 shows the compound detected at 0.869 min, panel 2 shows punicalagin, and panel 3 shows the compound detected at 7.509 and ellagic acid, respectively.

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  Fig. 5 Accurate mass spectra and chemical structure of the peaks detected in PV only. (A) compound detected at 0.869 min, (B) punicalagin B at 5.527 min, (C) compound detected at 7.509 min, and (D) ellagic acid ay 7.608 min.

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  Fig. 6 Comparisons of HPLC chromatograms for PV (Black filled) and pomegranate extract (Shaded). (A) shows the overlaid chromatogram of PV and pomegranate extract. HPLC chromatogram magnifications of panel 1 (B) and panel 2 (C) clearly show PV-specific peaks as indicated by down arrow in the chromatogram. Panel 1 shows the compound detected at 0.869 min and panel 2 shows punicalagin, the compound detected at 7.509 and ellagic acid, respectively.

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