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Aging Lowers Steady-State Antioxidant Enzyme and Stress Protein Expression in Primary Hepatocytes

^a Department of Exercise Science, The University of Iowa, Iowa City
^b Free Radical and Radiation Biology Program, The University of Iowa, Iowa City
^c McArdle Laboratory for Cancer Research, University of Wisconsin Medical School, Madison

Kevin C. Kregel, Department of Exercise Science, 532 FH, The University of Iowa, Iowa City, IA 52242 E-mail: kevin-kregel{at}uiowa.edu.

Decision Editor: John A. Faulkner, PhD

It has been reported that the isolation and culture of primary hepatocytes can compromise cellular ability to constituitively express antioxidant enzyme (AE) genes, making it difficult to study their regulation ex vivo. In the present study, the steady-state expression of manganese-containing superoxide dismutase, copper- and zinc-containing superoxide dismutase, catalase, and glutathione peroxidase was assessed in primary hepatocytes isolated from young and senescent rats and cultured in Matrigel. There was no change in steady-state superoxide dismutase protein or activity levels in cells collected from young animals and cultured for 7 days. Catalase expression was initially increased, and then it declined 30%. In contrast, superoxide dismutase expression declined 60% and catalase expression declined 50% in cells from senescent animals. Constitutive and inducible 70-kDa heat shock protein expression increased coincident with declining AE levels in the young cells but not senescent cells. For both age groups, electron micrographs showed rounded hepatocytes with abundant rough endoplasmic reticulum, mitochondria, and peroxisomes. Hepatocytes were organized into clusters of 6–12 cells surrounding a large central lumen devoid of microvilli. Each cluster also contained smaller microvilli-lined lumens between adjacent hepatocytes that resembled canniculi. The plasma membranes of these lumens were sealed from the extracellular space by junctional complexes. Gap junctions in the plasma membrane suggest that hepatocytes were capable of intercellular communication. We conclude that the Matrigel system can be used to study AE regulation in primary hepatocytes from young and senescent animals, provided that experiments can be conducted within a time frame of 5–7 days in culture. These data also support the hypothesis that aging compromises hepatocellular ability to maintain AE status and upregulate stress protein expression.

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				K. C. Kregel Molecular Biology of Thermoregulation: Invited Review: Heat shock proteins: modifying factors in physiological stress responses and acquired thermotolerance J Appl Physiol, May 1, 2002; 92(5): 2177 - 2186. [Abstract] [Full Text] [PDF]