Roles of multiple oxidized LDL lipids in cellular injury: dominance of 7 beta-hydroperoxycholesterol.

The relative toxicities of several lipid oxidation products formed on oxidized LDL, their presence on oxidized LDL, and potential mechanisms of cell injury compared to oxidized LDL were examined. Toxicities to fibroblasts, with lipoprotein-deficient serum supplementation, were: 7 beta-hydroperoxycholesterol > 7 beta-hydroxycholesterol = 4-hydroxynonenal > 7-ketocholesterol > 5 alpha, 6 alpha-epoxycholesterol. Lysophosphatidylcholine was only significantly cytotoxic in the absence of lipoprotein-deficient serum. Without serum, relative toxicities were: 7 beta-hydroperoxycholesterol > lysophosphatidylcholine > 4-hydroxynonenal > 7 beta-hydroxycholesterol. Similar relative potencies were observed in smooth muscle and endothelial cell cultures. 7 beta-Hydroperoxycholesterol accumulated on oxidized LDL to greater amounts than other oxysterols and 4-hydroxynonenal, but less than lysophosphatidylcholine. Cell injury by 7 beta-hydroperoxycholesterol and oxidized LDL was inhibitable by antioxidants but not by exogenous cholesterol or cycloheximide. In contrast, a) toxicities by 7 beta-hydroxycholesterol, 7-ketocholesterol, 5 alpha, 6 alpha-epoxycholesterol, and 4-hydroxynonenal were not inhibited by antioxidants; b) 7 beta-hydroxycholesterol and lysophosphatidylcholine toxicities were inhibited by exogenous cholesterol; and c) 7 beta-hydroxycholesterol toxicity was inhibited by cycloheximide. Injury by lysophosphatidylcholine was reduced by vitamin E and not affected by altering the cellular exposure to selenium; reduced selenium enhanced toxicity by oxidized LDL and 7 beta-hydroperoxycholesterol. The high relative toxicity of 7 beta-hydroperoxycholesterol, the level of its accumulation on oxidized LDL, and its mechanism of action similar to oxidized LDL suggest that it is the compound predominantly responsible for oxidized LDL induced cytotoxicity.