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A superfusion bioassay for platelet-activating factor.

G Cirino1, J L Wallace

  • 1Department of Pharmacology, University of Naples, Italy.

Canadian Journal of Physiology and Pharmacology
|January 1, 1989
PubMed
Summary
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A novel superfusion bioassay offers enhanced sensitivity for detecting platelet-activating factor (PAF) in various tissues. This method avoids desensitization, allowing for accurate measurement of even minute PAF quantities.

Area of Science:

  • Pharmacology
  • Biochemistry
  • Physiology

Background:

  • Platelet-activating factor (PAF) is a potent lipid mediator involved in various physiological and pathological processes.
  • Existing bioassays for PAF often lack sensitivity or suffer from tissue desensitization.
  • Accurate quantification of PAF is crucial for understanding its role in inflammation, thrombosis, and allergic responses.

Purpose of the Study:

  • To develop and characterize a sensitive superfusion bioassay for platelet-activating factor (PAF).
  • To evaluate the assay's performance across different tissue types and its ability to prevent desensitization.
  • To compare the novel bioassay with a conventional platelet aggregation assay for PAF measurement.

Main Methods:

  • A superfusion bioassay system was established using various isolated tissues.

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  • Tissues were washed with bovine serum albumin to prevent desensitization during repeated PAF additions.
  • The assay's sensitivity was determined, and PAF release from rat stomach samples was measured and correlated with a platelet aggregation bioassay.

    • Main Results:

    • The superfusion bioassay demonstrated high sensitivity, detecting PAF in the femtogram range (100-500 fg) with rat and dog ascending colon being the most responsive tissues.
    • Desensitization was minimal in most tissues, allowing for measurements over extended periods (up to 4 hours).
    • Excellent correlation (r = 0.96) was observed between the superfusion bioassay and a standard platelet aggregation assay for measuring PAF release.

    Conclusions:

    • The developed superfusion bioassay provides a highly sensitive, reproducible, and cost-effective method for quantifying platelet-activating factor.
    • This assay overcomes limitations of previous methods, such as tissue desensitization and lower sensitivity.
    • The superfusion bioassay represents a valuable alternative for researchers studying PAF in diverse biological contexts.