Effects of copper and histidine on oxidative modification of low density lipoprotein and its subsequent binding to collagen.
It was previously shown that serum low density lipoprotein (LDL) binds to type I collagen gels and that the binding is increased after modification by cultured endothelial cells. It is now demonstrated that when LDL is incubated with cells cultured in Dulbecco's modified minimal essential medium (DMEM), the subsequent binding of LDL to collagen is considerably less than after incubation with endothelial cells cultured in Ham's F-12 medium (F12). To determine the reason for this difference, collagen gels were made with saline containing ingredients of DMEM individually or in groups, and binding of LDL to such gels was measured. Modification of LDL, manifested by a high level of binding to the collagen, by lipid peroxidation (production of thiobarbituric acid-reactive substances), and by increased electrophoretic mobility occurred on exposure to collagen gels made in saline; these changes were almost completely inhibited by the addition of histidine at a concentration equal to that in DMEM. They were also inhibited by butylated hydroxytoluene, desferrioxamine, and EDTA; penicillamine and hydroxyl-radical scavengers inhibited collagen binding but did not inhibit lipid peroxidation or the increase in electrophoretic mobility. Nominally, DMEM contains 270 microM histidine but no copper, whereas F12 contains 135 microM histidine and 10 nM copper; addition of copper (as much as 5 microM) to DMEM or of histidine (as much as 2.16 mM) to F12 did not overcome the differences between the media in supporting LDL oxidation by endothelial cells.(ABSTRACT TRUNCATED AT 250 WORDS)
Kalant
N
N
Department of Medicine, McGill University, Montreal, Canada.
McCormick
S
S
Parniak
M A
MA
D000818:Animals; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism*; D003300:Copper / Q000494:pharmacology*; D003470:Culture Media; D004730:Endothelium, Vascular / Q000187:drug effects / Q000378:metabolism; D006639:Histidine / Q000494:pharmacology*; D015227:Lipid Peroxidation / Q000187:drug effects; D008077:Lipoproteins, LDL / Q000378:metabolism*; D010084:Oxidation-Reduction; D011817:Rabbits
D003470:Culture Media; D008077:Lipoproteins, LDL; D006639:Histidine; D003300:Copper; D003094:Collagen