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The effect of conjugated linoleic acid on the antioxidant enzyme defense system in rat hepatocytes

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Lipids

Abstract

Short-term effects of physiological concentrations of conjugated linoleic acid (CLA) on membrane integrity, metabolic function, cellular lipid composition, lipid peroxidation, and antioxidant enzymes were examined using rat hepatocyte suspension cultures. Incubation with CLA (5–20 ppm) for 3 h decreased the ability of hepatocyte plasma membranes to exclude trypan blue by approximately 25%, and caused leakage of cytosolic lactate dehydrogenase (LDH) into the medium. The significant decrease (P<0.02) in hepatocyte viability as measured by LDH leakage during cell incubation with 10 and 20 ppm CLA was not associated with significant changes in cellular ATP content. Protein synthesis in hepatocytes was elevated (P<0.05) in the presence of 5 and 10 ppm CLA, but at a higher concentration (20 ppm), protein synthesis was similar to that of control cells. Gluconeogenesis was maintained in cells incubated with lower concentrations of CLA (5 and 10 ppm) but was decreased (P<0.02) at the higher concentration. Incubation with 20 ppm CLA for 3 h did not affect the specific activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase, the rate-limiting enzyme of cholesterol synthesis. Both cis-9,trans-11/trans-9,cis-11, and cis-10,trans-12/trans-10,cis-12 isomers of CLA were incorporated to a similar level into hepatocytes. Levels ranged from 3.9 to 4.1%, respectively, of total fatty acids in neutral lipids, and from 0.7 to 0.8%, respectively, of total fatty acids in phospholipids. Cellular lipid peroxidation remained unchanged in the presence of CLA (5–20 ppm), despite significant inhibition (P<0.05) of superoxide dismutase. Catalase activity was maintained near control levels in the presence of 5 and 10 ppm CLA but was significantly decreased in the presence of 20 ppm CLA. Glutathione peroxidase activity was significantly decreased in the presence of 10 ppm CLA. The apparent sensitivity of the antioxidant enzyme defense system of liver cells to CLA, coupled with the lack of effect of CLA on lipid peroxidation in cells, suggests that cytotoxic effects of CLA as described by LDH leakage and decreased gluconeogenesis were not mediated by a prooxidant action in hepatocytes.

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Abbreviations

CLA:

conjugated linoleic acid

FAME:

fatty acid methyl ester

GLC:

gas-liquid chromatography

GPx:

gluthathione peroxidase

HBSS:

Hanks' balanced salt solution

HMGCoA reductase:

3-hydroxy-3-methylglutaryl coenzyme A reductase

LDH:

lactate dehydrogenase

PBS:

phosphate-buffered saline

PUFA:

polyunsaturated fatty acids

SOD:

superoxide dismutase

TBARS:

thiobarbituric acid-reactive substances

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Authors and Affiliations

  1. School of Biotechnology, Dublin City University, Dublin 9, Ireland

    H. Cantwell, R. Devery & M. O'Shea

  2. Teagasc, Dairy Products Research Centre, Moorepark, Fermoy County Cork, Ireland

    M. O'Shea & C. Stanton

Authors
  1. H. Cantwell
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  2. R. Devery
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  3. M. O'Shea
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  4. C. Stanton
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Corresponding author

Correspondence to R. Devery.

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Cantwell, H., Devery, R., O'Shea, M. et al. The effect of conjugated linoleic acid on the antioxidant enzyme defense system in rat hepatocytes. Lipids 34, 833–839 (1999). https://doi.org/10.1007/s11745-999-0430-4

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  • DOI: https://doi.org/10.1007/s11745-999-0430-4

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