Luminol-dependent chemiluminescence analysis of human platelets |
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Authors: | Knox Van Dyke Cynthia Van Dyke David Peden George Jones Vincent Castranova Eric Brestel Michael Ringrose |
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Affiliation: | 1. Departments of Pharmacology and Toxicology, West Virginia University Medical Center, Morgantown, West Virginia 26506 U.S.A.;2. Department of Physiology, (ALOSH, NIOSH, CDC and HEW), Morgantown, West Virginia 26506 U.S.A.;3. Department of Medicine-Allergy Section, West Virginia University Medical Center, Morgantown, West Virginia 26506 U.S.A.;4. Analytical Luminescence Laboratory, West Lake Village, California 91361 U.S.A. |
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Abstract: | We describe a simple, sensitive, and reproducible assay system to measure the chemiluminescence (CL) produced by injecting arachidonic acid (AA) into a preparation of human platelets containing luminol.The CL appears to result from the metabolism of the AA by enzymes in human platelets, namely, cyclooxygenase, lipoxygenase, and possibly peroxidases. It is believed that when the AA is injected, free radicals and/or oxidizing agents are formed that react with the luminol producing an excited state and emitting blue light (425 nm).The enzymes can be inhibited by drugs to varying degrees. BW755C inhibits all CL at micromolar doses and it is known to inhibit both lipoxygenase and cyclooxygenase. Aspirin, indomethacin, and sulindac sulfide inhibit only cyclooxygenase and inhibit 35–65% of the light from an individual. This assay system can be used to screen certain drugs that are effective in inflammatory diseases. It could be used to determine whether the drugs would be effective in a given individual and also whether drugs have a long-term toxic effect in vivo on platelets. Further the assay is practical with a few milliters of blood. |
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