Biochemical and 31P-NMR spectroscopic evaluation of immobilized perifused rat hepatocytes exposed to anoxia: Relation to the nutritional status

A. Gasbarrini, A. B. Borle, P. Caraceni, H. Farghali, A. Gasbarrini, R. Giardino, A. Facchini, M. Bernardi, D. H. Van Thiel

Research output: Contribution to journalArticlepeer-review

Abstract

The effects of anoxia on ATP, cytosolic free calcium concentration and cell injury were studied in hepatocytes isolated from fed or 24 hrs fasted rats. After isolation, the cells were imbedded in agarose gel threads and continuously perfused with oxygenated Krebs-Henseleit bicarbonate buffer (KHB). Anoxia was obtained exposing the cells for 2 hrs to 95% N2-5% CO2. Cytosolic free calcium (Ca(i)2+) was measured in real time with the photoprotein aequorin; ATP (expressed as the β-ATP/external standard ratio) by 31P-NMR spectroscopy; and cytotoxicity by measuring the LDH released by the cells. In control conditions, hepatocytes from fasted rats showed a 48% decrease in ATP (p <0.01) and a 35% increase in Ca(i)2+ (p2+ increased faster and to significantly higher levels than in hepatocytes from fed rats: Ca(i)2+ reached 2.1±0.2 μM within 10 min compared to 1.45±0.1 μM in 1 hr, respectively (p2+ increase and LDH release was r2=0.95. These results show that anoxic injury is markedly increased after fasting and that there is a significant correlation between the rise in Ca(i)2+ evoked by anoxia and the cell damage.

Original languageEnglish
Pages (from-to)156-159
Number of pages4
JournalTransplantationsmedizin: Organ der Deutschen Transplantationsgesellschaft
Volume6
Issue number2
Publication statusPublished - 1994

ASJC Scopus subject areas

  • Immunology and Allergy
  • Transplantation

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