Antioxidative function of L-FABP in L-FABP stably transfected Chang liver cells

Hepatology. 2005 Oct;42(4):871-9. doi: 10.1002/hep.20857.


Liver fatty acid binding protein (L-FABP) contains amino acids that are known to possess antioxidant function. In this study, we tested the hypothesis that L-FABP may serve as an effective endogenous cytoprotectant against oxidative stress. Chang liver cells were selected as the experimental model because of their undetectable L-FABP mRNA level. Full-length L-FABP cDNA was subcloned into the mammalian expression vector pcDNA3.1 (pcDNA-FABP). Chang cells were stably transfected with pc-DNA-FABP or vector (pcDNA3.1) alone. Oxidative stress was induced by incubating cells with 400 micromol/L H2O2 or by subjecting cells to hypoxia/reoxygenation. Total cellular reactive oxygen species (ROS) was determined using the fluorescent probe DCF. Cellular damage induced by hypoxia/reoxygenation was assayed by lactate dehydrogenase (LDH) release. Expression of L-FABP was documented by regular reverse transcription polymerase chain reaction (RT-PCR), real-time RT-PCR, and Western blot. The pcDNA-FABP-transfected cells expressed full-length L-FABP mRNA, which was absent from vector-transfected control cells. Western blot showed expression of 14-kd L-FABP protein in pcDNA-FABP-transfected cells, but not in vector-transfected cells. Transfected cells showed decreased DCF fluorescence intensity under oxidative stress (H2O2 and hypoxia/reoxygenation) conditions versus control in inverse proportion to the level of L-FABP expression. Lower LDH release was observed in the higher L-FABP-expressed cells in hypoxia/reoxygenation experiments. In conclusion, we successfully transfected and cloned a Chang liver cell line that expressed the L-FABP gene. The L-FABP-expressing cell line had a reduced intracellular ROS level versus control. This finding implies that L-FABP has a significant role in oxidative stress.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antioxidants / metabolism*
  • Apoptosis
  • Carcinoma, Hepatocellular
  • Carrier Proteins / genetics*
  • Carrier Proteins / metabolism*
  • Cell Line, Tumor
  • DNA, Complementary
  • Fatty Acid-Binding Proteins
  • Glutathione / metabolism
  • Hepatocytes / cytology
  • Hepatocytes / metabolism*
  • Hepatocytes / physiology
  • Humans
  • Hydrogen Peroxide / pharmacology
  • Hypoxia / metabolism
  • Liver Neoplasms
  • Oxidants / pharmacology
  • Oxidative Stress / drug effects
  • Oxidative Stress / physiology
  • Reactive Oxygen Species / metabolism
  • Reperfusion Injury / metabolism
  • Superoxide Dismutase / metabolism
  • Transfection / methods*
  • Tumor Necrosis Factor-alpha / metabolism


  • Antioxidants
  • Carrier Proteins
  • DNA, Complementary
  • FABP1 protein, human
  • Fatty Acid-Binding Proteins
  • Oxidants
  • Reactive Oxygen Species
  • Tumor Necrosis Factor-alpha
  • Hydrogen Peroxide
  • Superoxide Dismutase
  • Glutathione