J Korean Med Sci.  2012 Jun;27(6):644-652. 10.3346/jkms.2012.27.6.644.

Bisphenol A Impairs Mitochondrial Function in the Liver at Doses below the No Observed Adverse Effect Level

• Moon MK
• Kim MJ
• Jung IK
• Koo YD
• Ann HY
• Lee KJ
• Kim SH
• Yoon YC
• Cho BJ
• Park KS
• Jang HC
• Park YJ

• KMID: 1421617
• DOI: http://doi.org/10.3346/jkms.2012.27.6.644

Abstract

Bisphenol A (BPA) has been reported to possess hepatic toxicity. We investigated the hypothesis that BPA, below the no observed adverse effect level (NOAEL), can induce hepatic damage and mitochondrial dysfunction by increasing oxidative stress in the liver. Two doses of BPA, 0.05 and 1.2 mg/kg body weight/day, were administered intraperitoneally for 5 days to mice. Both treatments impaired the structure of the hepatic mitochondria, although oxygen consumption rate and expression of the respiratory complex decreased only at the higher dose. The hepatic levels of malondialdehyde (MDA), a naturally occurring product of lipid peroxidation, increased, while the expression of glutathione peroxidase 3 (GPx3) decreased, after BPA treatment. The expression levels of proinflammatory cytokines such as interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) also increased. In HepG2 cells, 10 or 100 nM of BPA also decreased the oxygen consumption rate, ATP production, and the mitochondrial membrane potential. In conclusion, doses of BPA below the NOAEL induce mitochondrial dysfunction in the liver, and this is associated with an increase in oxidative stress and inflammation.

Keyword

Bisphenol A; Liver; Mitochondria; Oxidative Stress; Inflammation

MeSH Terms

Adenosine Triphosphate/metabolism
Animals
Glutathione Peroxidase/metabolism
Hep G2 Cells
Humans
Inflammation/chemically induced/metabolism/pathology
Injections, Intraperitoneal
Interleukin-6/metabolism
Lipid Peroxidation/drug effects
Liver/*drug effects/metabolism/pathology
Male
Malondialdehyde/metabolism
Membrane Potential, Mitochondrial/drug effects
Mice
Mice, Inbred C57BL
Mitochondria/drug effects/*metabolism
Oxidative Stress/drug effects
Oxygen Consumption/drug effects
Phenols/*toxicity
Tumor Necrosis Factor-alpha/metabolism
Interleukin-6
Phenols
Tumor Necrosis Factor-alpha
Malondialdehyde
Adenosine Triphosphate
Glutathione Peroxidase

Figure

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  Fig. 1 Structure and function of hepatic mitochondria in the mice treated with 1.2 mg/kg bw/day of BPA for 5 days. (A) Morphology of hepatic mitochondria by transmission electron microscopy (150,000 × magnification). Control mice (A1) and BPA-treated mice (A2). Normal mitochondria (arrows) and swollen mitochondria (arrowheads). (B) Oxygen consumption rate of hepatic mitochondria. GMD, glutamate + malate + ADP; GMcD, GMD + cytochrome c; GMcDS, GMcD + succinate; FCCP, uncoupler (*P < 0.05 compared to control). (C) The respiratory complex I-V of hepatic mitochondria.

• 

  Fig. 2 Serum AST and ALT levels after a single injection of 1.2 mg/kg bw/day of BPA.

• 

  Fig. 3 Changes in the levels of oxidative stress and inflammatory cytokines at 1, 6, and 24 hr after a single injection of BPA (1.2 mg/kg bw/day). (A) MDA concentrations in the liver. (B) Catalase and GPx3 in the liver. (C) IL-6 and TNF-α levels in the liver. (D) Serum IL-6 and TNF-α levels. (E) Oxygen consumption rate in hepatic mitochondria. GMD, glutamate + malate + ADP; GMcD, GMD + cytochrome c; GMcDS, GMcD + succinate (*P < 0.05 compared to control).

• 

  Fig. 4 Oxidative stress in the liver of mice treated with BPA (1.2 mg/kg bw/day) for 5 days. (A) MDA concentrations (*P < 0.05 compared to control). (B) Catalase and GPx3 in the liver.

• 

  Fig. 5 Structure and function of hepatic mitochondria in the mice treated with BPA (0.05 mg/kg bw/day) for 5 days. (A) Morphology of hepatic mitochondria by transmission electron microscopy (500,000 × magnification). Control (A1) and BPA-treated mice (A2, A3). Normal mitochondria (arrows) and swollen and cristae-disrupted mitochondria (arrowhead). (B) Oxygen consumption rate of hepatic mitochondria. GMD, glutamate + malate + ADP; GMcD, GMD + cytochrome c; GMcDS, GMcD + succinate; FCCP, uncoupler. (C) MDA concentrations in the liver. (D) Catalase and GPx3 in the liver.

• 

  Fig. 6 Structure and function of the mitochondria in HepG2 cells treated with BPA. (A) Mitochondrial morphology of HepG2 cells by transmission electron microscopy (300,000 × magnification). Control HepG2 cells (A1), 10 nM BPA-treated HepG2 cells (A2), and 100 nM BPA-treated HepG2 cells (A3). Normal mitochondria (arrows) and swollen and cristae-disrupted mitochondria (arrowheads). (B) Oxygen consumption rate of HepG2 cells. GMD, glutamate + malate + ADP; GMcD, GMD + cytochrome c; GMcDS, GMcD + succinate; FCCP, uncoupler. (C) ATP production after the treatment with 100 nM BPA. (D) MMP after the treatment with 10 or 100 nM BPA (*P < 0.05 compared to control; †P < 0.01 compared to control).

• 

  Fig. 7 Oxidative stress in HepG2 cells treated with 100 nM BPA. (A) MDA concentrations determined by a TBARS assay kit. (B) Fluorescence after the ROS-sensitive indicator DHE staining. Control (B1), 2 hr (B2), 6 hr (B3), and 24 hr (B4) after the treatment of 100 nM BPA.

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