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 language | English (US) |
|---|---|
| Pages (from-to) | 105-114 |
| Number of pages | 10 |
| Journal | BBA - Molecular Basis of Disease |
| Volume | 1139 |
| Issue number | 1-2 |
| DOIs | |
| State | Published - Jun 9 1992 |
| Externally published | Yes |
Fingerprint
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 journal › Article
}
TY - JOUR
T1 - 31P-NMR spectroscopy of perifused rat hepatocytes immobilized in agarose threads
T2 - application to chemical-induced hepatotoxicity
AU - Farghali, Hassan
AU - Rossaro, Lorenzo
AU - Gavaler, Judith S.
AU - H. Van Thiel, David
AU - Dowd, Susan R.
AU - Williams, Donald S.
AU - Ho, Chien
PY - 1992/6/9
Y1 - 1992/6/9
N2 - 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.
AB - 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.
KW - P-NMR
KW - Acetaminophen
KW - Agarose thread
KW - Allyl alcohol
KW - Ethanol
KW - Fructose
KW - Hepatocyte
KW - Hepatotoxicity
UR - http://www.scopus.com/inward/record.url?scp=0027115835&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0027115835&partnerID=8YFLogxK
U2 - 10.1016/0925-4439(92)90089-6
DO - 10.1016/0925-4439(92)90089-6
M3 - Article
VL - 1139
SP - 105
EP - 114
JO - Biochimica et Biophysica Acta - Molecular Basis of Disease
JF - Biochimica et Biophysica Acta - Molecular Basis of Disease
SN - 0925-4439
IS - 1-2
ER -