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Related Experiment Videos

Changes in hepatocyte NADH fluorescence during prolonged hypoxia

E T Obi-Tabot1, L M Hanrahan, R Cachecho

  • 1Surgical Research Section, Boston University School of Medicine, Massachusetts 02118.

The Journal of Surgical Research
|December 1, 1993
PubMed
Summary

Monitoring cellular NADH fluorescence can track tissue damage during prolonged hypoxia. Early increases indicate impaired energy production, while later declines signal irreversible cell injury, crucial for understanding ischemia.

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Area of Science:

  • Biochemistry
  • Physiology
  • Medical Imaging

Background:

  • Hypoxia impairs oxidative phosphorylation, increasing cellular NADH.
  • NADH fluorescence monitoring is established for acute hypoxia.
  • Clinical relevance of NADH monitoring during prolonged ischemia is unexplored.

Purpose of the Study:

  • To investigate NADH fluorescence changes during extended hypoxia in liver cells and perfused livers.
  • To assess the correlation between NADH fluorescence and cell viability during ischemia.
  • To evaluate fluorimetry's utility for monitoring clinical ischemia.

Main Methods:

  • Studied rat hepatocyte suspensions and isolated-perfused rat livers.
  • Induced hypoxia (pO2 < 50 mm Hg) for up to 180 minutes.

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  • Monitored NADH fluorescence using fluorimetry and fiberoptic probes.
  • Assessed cell viability via trypan blue exclusion and ATP levels.

    • Main Results:

    • Hepatocytes showed a 15% NADH increase within 15 min, coinciding with ATP decrease.
    • Short hypoxia (15-30 min) with reoxygenation allowed ATP/NADH recovery and maintained viability.
    • Prolonged hypoxia (60-180 min) led to an initial NADH rise followed by an irreversible decline, correlating with cell death.
    • Perfused livers exhibited delayed NADH increases after 45-60 min, peaking and then declining irreversibly with extended hypoxia.

    Conclusions:

    • NADH fluorescence monitoring can track both reversible and irreversible changes during prolonged liver ischemia.
    • Early NADH increases reflect metabolic stress, while subsequent declines indicate cell injury.
    • Fluorimetry holds potential for assessing tissue status during extended clinical ischemic events.