31P-NMR spectroscopy of perifused rat hepatocytes immobilized in agarose threads

application to chemical-induced hepatotoxicity

Hassan Farghali, Lorenzo Rossaro, Judith S. Gavaler, David H. Van Thiel, Susan R. Dowd, Donald S. Williams, Chien Ho

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

A system consisting of isolated rat hepatocytes immobilized in agarose threads continuously perifused with oxygenated Krebs-Henseleit (KH) solution has been found to maintain cell viability with excellent metabolic activity for more than 6 h. The hepatocytes were monitored by phosphorus-31 nuclear magnetic resonance (31P-NMR) spectroscopy at 4.7 Tesla, by measurement of oxygen consumption and by the leakage of lactate dehydrogenase (LD) and alanine aminotransferase (ALT). The data obtained were comparable to those found for an isolated perfused whole liver in vitro. The effects of allyl alcohol (AA), ethanol, and 4-acetaminophenol (AP) were examined. A solution of 225 μM AA perifused for 90 min caused the disappearance of the β-phosphate resonance of adenosine triphosphate (ATP) in the 31P-NMR spectra, a 7-fold increase in LD leakage and a 70% reduction in oxygen consumption. Ethanol (1.0 M) perifused for 90 min reduced the β-ATP signal intensity ratio by 20%, the phosphomonoester (PME) signal by 50% and inorganic phosphate (Pi) by 33% (P < 0.05). AP (10 mM) caused only mild liver-cell damage. The results demonstrate that perifused immobilized hepatocytes can be used as a liver model to assess the effects of a wide range of chemicals and other xenobiotics by NMR spectroscopy.

Original languageEnglish (US)
Pages (from-to)105-114
Number of pages10
JournalBBA - Molecular Basis of Disease
Volume1139
Issue number1-2
DOIs
StatePublished - Jun 9 1992
Externally publishedYes

Fingerprint

Sepharose
Hepatocytes
Magnetic Resonance Spectroscopy
L-Lactate Dehydrogenase
Oxygen Consumption
Liver
Ethanol
Adenosine Triphosphate
Phosphates
Xenobiotics
Alanine Transaminase
Phosphorus
Cell Survival
allyl alcohol
In Vitro Techniques
Krebs-Henseleit solution

Keywords

  • P-NMR
  • Acetaminophen
  • Agarose thread
  • Allyl alcohol
  • Ethanol
  • Fructose
  • Hepatocyte
  • Hepatotoxicity

ASJC Scopus subject areas

  • Biophysics
  • Molecular Biology
  • Molecular Medicine
  • Medicine(all)

Cite this

31P-NMR spectroscopy of perifused rat hepatocytes immobilized in agarose threads : application to chemical-induced hepatotoxicity. / Farghali, Hassan; Rossaro, Lorenzo; Gavaler, Judith S.; H. Van Thiel, David; Dowd, Susan R.; Williams, Donald S.; Ho, Chien.

In: BBA - Molecular Basis of Disease, Vol. 1139, No. 1-2, 09.06.1992, p. 105-114.

Research output: Contribution to journalArticle

Farghali, Hassan ; Rossaro, Lorenzo ; Gavaler, Judith S. ; H. Van Thiel, David ; Dowd, Susan R. ; Williams, Donald S. ; Ho, Chien. / 31P-NMR spectroscopy of perifused rat hepatocytes immobilized in agarose threads : application to chemical-induced hepatotoxicity. In: BBA - Molecular Basis of Disease. 1992 ; Vol. 1139, No. 1-2. pp. 105-114.
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