Oxysterols and TBARS are among the LDL oxidation products which enhance thromboxane A2 synthesis by platelets.
In this study, we compared the effects of normal LDL (nLDL) and oxidized LDL (oxLDL) on thromboxane (TXA2) release by platelets triggered by low concentration of thrombin, and we determined which component of oxLDL is responsible for that activation. After oxidation of LDL with copper sulfate, the small molecular weight fraction (< 10 kDa) which was high in TBARS was removed; using Amicon Centriprep-10 concentrator membrane. More than 67% of TBARS in the oxLDL preparation was found in solution while the remaining was covalently attached to the oxLDL particles. OxLDL contained significantly higher levels of oxysterols and TBARS than the nLDL. Platelets preincubated with low concentrations of oxLDL (33-132 micrograms protein/mL) produced significantly higher TXA2 than platelets preincubated with equivalent concentrations of nLDL when triggered with thrombin. Platelets treated with oxLDL also contained significantly higher levels of oxysterols than platelets treated with nLDL. Platelets preincubated with pure cholestanetriol (10 micrograms/mL) contained a high level of cholestanetriol in the membrane, and TXA2 release was significantly increased in these platelets compared to the control platelets. The TBARS in solution also was very potent in enhancing TXA2 release by thrombin-treated platelets. These results indicate that oxysterols and the free TBARS either in solution or covalently attached to the oxLDL particles are partly responsible for the stimulatory effect of oxLDL on TXA2 release by platelets. The present study also showed that this enhancement of TXA2 release was due to activation of phospholipase A2 and to the increase of arachidonic acid liberation from the platelet phospholipids.
Mahfouz
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MM
University of Illinois, Burnsides Research Laboratory, Urbana 61801, USA. fkummerow@uiuc.edu
Kummerow
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FA
D000475:Alkenes / Q000378:metabolism; D016718:Arachidonic Acid / Q000378:metabolism / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002250:Carbon Radioisotopes; D002777:Cholestanols / Q000378:metabolism / Q000494:pharmacology; D002784:Cholesterol / Q000032:analysis / Q000378:metabolism; D006801:Humans; D008077:Lipoproteins, LDL / Q000378:metabolism* / Q000494:pharmacology; D010084:Oxidation-Reduction; D013261:Sterols / Q000378:metabolism*; D017392:Thiobarbituric Acid Reactive Substances / Q000378:metabolism*; D013917:Thrombin / Q000494:pharmacology; D013928:Thromboxane A2 / Q000096:biosynthesis*; D014810:Vitamin E / Q000378:metabolism / Q000494:pharmacology
D000475:Alkenes; D002250:Carbon Radioisotopes; D002777:Cholestanols; D008077:Lipoproteins, LDL; D013261:Sterols; D017392:Thiobarbituric Acid Reactive Substances; C000474:cholestane-3,5,6-triol; D014810:Vitamin E; D016718:Arachidonic Acid; D013928:Thromboxane A2; D002784:Cholesterol; D013917:Thrombin