A new method for determining cellular ATP content in hepatocytes.
We have developed a new ATP assay for hepatocytes which is simple, rapid and highly sensitive. ATP determination using luciferin-luciferase was performed on hepatocytes separated by perfusion of the liver with collagenase. There was a close correlation between the ATP content of hepatocytes and viable cell numbers. The ATP content of dead cells which were determined by trypan blue dye exclusion test had less than one per cent of levels of viable cells. ATP contents of isolated hepatocytes in Minimum Essential Medium was 6.8 +/- 0.6 x 10(-15) mol/cell at 2 hours after excision of the liver and showed no significant difference compared with that determined at 6 hours. This method was performed to evaluate changes in the cellular ATP content of hepatocytes after partial hepatectomy in rats and transcatheter portal embolization (TPE) in dogs. The ATP content in rat hepatocytes showed a remarkable increase after hepatectomy, with a peak value of 19.1 +/- 1.7 x 10(-15) mol/cell at 24 hours post-surgery. On the other hand, marked atrophy in the embolized lobes and compensatory hypertrophy in the non-embolized lobes were found following TPE in dogs. Cellular ATP content in the non-embolized lobes showed its highest level of 8.7 +/- 2.9 x 10(-15) mol/cell on the third day after TPE, but in the embolized lobes decreased immediately after TPE with significant differences compared with the non-embolized lobes (p < 0.05). Our method may also be applicable to the evaluation of other adenosine phosphate by use of converting enzymes for ATP.(ABSTRACT TRUNCATED AT 250 WORDS)
Ishikawa
M
M
1st Department of Surgery, School of Medicine, University of Tokushima, Japan.
Yogita
S
S
Takai
S
S
Komi
N
N
Makuuchi
M
M
D000255:Adenosine Triphosphate / Q000032:analysis*; D000818:Animals; D001681:Biological Assay / Q000379:methods*; D004285:Dogs; D004617:Embolism; D049454:Firefly Luciferin; D006498:Hepatectomy; D008099:Liver / Q000737:chemistry* / Q000166:cytology; D008156:Luciferases; D008163:Luminescent Measurements; D051381:Rats; D017208:Rats, Wistar; D013997:Time Factors
D049454:Firefly Luciferin; D000255:Adenosine Triphosphate; D008156:Luciferases