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Oxidized low-density lipoproteins induce apoptosis in aortic and endocardial endothelial cells
1Laboratory of Molecular Pharmacology, Hunan Medical University, Changsha, China. guozg@public.cs.hn.cn
Aim:
To examine whether oxidized low-density lipoproteins (ox-LDL) might induce apoptosis in bovine aortic and endocardial endothelial cells (BAEC and BEEC).
Methods:
Low-density lipoproteins (LDL) were isolated from healthy human plasma by ultracentrifugation and oxidized by CuSO4 10 mumol.L-1. BAEC and BEEC were incubated in a medium containing ox-LDL, LDL, or phosphate-buffer solution (PBS) as control. DNA fragmentation was visualized by agarose gel electrophoresis and determined quantitatively using Hoechst-33258 fluorochrome.
Results:
Ox-LDL, not LDL, elicited typical apoptotic changes and DNA fragmentation in BAEC and BEEC. In BAEC, dextran sulfate, and cicloheximide (Cic) exhibited no effect on DNA fragmentation induced by ox-LDL. Butylated hydroxytoluene (BHT) 20 mumol.L-1 completely inhibited Cu(2+)-mediated oxidation of LDL as well as the apoptosis-inducing effect of Cu(2+)-exposed LDL. Lysophosphatidylcholine (LPC) did not elicit DNA fragmentation in BAEC and in BEEC. DNA fragmentation induced by ox-LDL in BAEC and in BEEC was blocked by chelating the calcium of the culture medium by egtazic acid.
Conclusion:
Ox-LDL induces apoptosis in BAEC and BEEC without involving the LPC.