Failure of vitamin E to protect cultured human arterial smooth muscle cells against oxysterol-induced cytotoxicity.
OBJECTIVE
The cytotoxicity of oxysterols including 7 alpha-hydroxycholesterol (7 alpha OHC), 7 beta-hydroxycholesterol (7 beta OHC), cholesterol 5 alpha,6 alpha-epoxide (alpha epoxyC), cholesterol 5 beta,6 beta-epoxide (beta epoxyC), 7-ketocholesterol (7ketoC), 26-hydroxycholesterol (26OHC), cholesterol-3 beta,5 alpha,6 beta-triol (TriolC) and the possible protecting effect of vitamin E on 26OHC-induced cytotoxicity were investigated in smooth muscle cells isolated from the arteries of human umbilical cords.


METHODS
To study the cytotoxicity of oxysterols, the cells were incubated with each oxysterol at a level of 10 micrograms/ml from 24 to 120 hours, then 45Ca++ uptake, cytosolic free Ca++ level, [3H]thymidine incorporation, total DNA content and viable cell number were measured. Cholesterol was used as a control. For tracing the possible origin of cytotoxicity of 26OHC, cholesterol, phospholipid and 26OHC content in the membrane were investigated from 24 to 72 hours. For determining whether antioxidants had a protective effect against the cytotoxicity of 26OHC, vitamin E and butylated hydroxytoluene (BHT) were used.


RESULTS
The results indicated that the oxysterols elevated 45Ca++ uptake and cytosolic free Ca++ level, but diminished [3H]thymidine incorporation, total DNA content and viable cell number. 26OHC lowered the cholesterol content of the membrane and incorporated into the membrane after 24 hours of the incubation, but did not alter the total phospholipid content of the membrane until 72 hours. Neither vitamin E or BHT significantly protected the cells from the 26OHC-induced alterations.


CONCLUSIONS
We suggest that the cytotoxicity of oxysterols, which might result in an alteration in Ca++ ion flow into the cell by decreasing cholesterol content and incorporating oxysterol itself into the membranes, could not be protected by vitamin E.
Zhou
Q
Q
Burnsides Research Laboratory, University of Illinois, Urbana 61801, USA.
Wasowicz
E
E
Kummerow
F A
FA
D002118:Calcium / Q000378:metabolism; D002132:Calcium Radioisotopes; D016923:Cell Death / Q000187:drug effects*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002478:Cells, Cultured; D002784:Cholesterol / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D004247:DNA / Q000096:biosynthesis; D006801:Humans; D006888:Hydroxycholesterols / Q000494:pharmacology*; D008563:Membrane Lipids / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D014469:Umbilical Arteries; D014810:Vitamin E / Q000494:pharmacology*
D002132:Calcium Radioisotopes; D006888:Hydroxycholesterols; D008563:Membrane Lipids; C007042:cholest-5-ene-3 beta,26-diol; D014810:Vitamin E; D004247:DNA; D002784:Cholesterol; D002118:Calcium