The dynamic reduction of Cu(II) to Cu(I) and not Cu(I) availability is a sufficient trigger for low density lipoprotein oxidation.
Copper (II) is one of the most widely employed experimental models to induce in vitro low density lipoprotein (LDL) oxidation. It is generally assumed that Cu(I), generated from active reduction of Cu(II), is the real trigger for the peroxidation of polyunsaturated fatty acids in LDL. We have employed isolated human LDL challenged with Cu(II) and the Cu(I) chelator bathocuproine disulfonic acid (BC) to test the validity of this hypothesis. At lower Cu(II)/LDL molar ratios, BC completely inhibited copper-mediated LDL oxidation evaluated either as thiobarbituric acid reactive substances (TBARS) production or changes in apo B100 electrophoretic mobility. On the contrary, at higher Cu(II)/LDL molar ratios, BC did not prevent LDL oxidation but rather markedly stimulated both the generation of TBARS and the increase of apo B100 electronegativity. These oxidative modifications were completely prevented by the Cu(II) chelator EDTA. Furthermore, the BC-Cu(I) complex alone was neither redox active nor active inducer of TBARS generation. These findings indicate that, under these experimental conditions, [1] Cu(I) is not necessary to promote LDL oxidation, [2] the availability of Cu(II) is a prerequisite and [3] some of the reaction(s) involved in Cu(II) reduction may play an essential role in initiating LDL oxidation.
Perugini
C
C
Department of Medical Sciences, University of Torino, Novara, Italy.
Seccia
M
M
Albano
E
E
Bellomo
G
G
D000328:Adult; D002614:Chelating Agents; D003300:Copper / Q000737:chemistry*; D019327:Copper Sulfate; D006801:Humans; D008077:Lipoproteins, LDL / Q000737:chemistry* / Q000302:isolation & purification; D010084:Oxidation-Reduction; D010618:Phenanthrolines; D017392:Thiobarbituric Acid Reactive Substances / Q000032:analysis; D013997:Time Factors
D002614:Chelating Agents; D008077:Lipoproteins, LDL; D010618:Phenanthrolines; D017392:Thiobarbituric Acid Reactive Substances; C028559:bathocuproine sulfonate; D003300:Copper; D019327:Copper Sulfate