Effect of 25-hydroxycholesterol, 26-hydroxycholesterol and its analogue, 26-aminocholesterol, on protein content, protein synthesis and LDH leakage in mouse epidermal cells.
The cytotoxicity of 25-hydroxycholesterol, 26-hydroxycholesterol and its analogue, 26-aminocholesterol was investigated in murine epidermal cell line, HEL-30. Lactate dehydrogenase (LDH) leakage, protein synthesis and protein content were determined after exposure of cell monolayers to the compounds ranging from 0.1-200 microM for 2, 6, or 24 h. 26-Aminocholesterol affected all the parameters studied time and concentration dependently; 25-hydroxycholesterol and 26-hydroxycholesterol were not toxic for HEL-30 cells. The cellular target for 26-aminocholesterol was primarily the membrane, since the LDH leakage was already detectable 10 min after exposure. On the other hand, for the other two oxysterols a protective role on this structure might be postulated. In fact 25-hydroxycholesterol and 26-hydroxycholesterol decreased the natural LDH leakage due to the ageing of the culture. In addition, 20 microM 25-hydroxycholesterol reversed the effect of moderately lytic doses of 26-aminocholesterol and Triton X-100, but not of sodium dodecyl sulfate.
Marinovich
M
M
Laboratory of Toxicology, Institute of Pharmacological Sciences, Milano, Italy.
Guizzetti
M
M
Ghilardi
F
F
Paoletti
R
R
Galli
C L
CL
D000704:Analysis of Variance; D000818:Animals; D015415:Biomarkers / Q000032:analysis; D002460:Cell Line; D002784:Cholesterol / Q000031:analogs & derivatives* / Q000633:toxicity; D004817:Epidermis; D006888:Hydroxycholesterols / Q000633:toxicity*; D007770:L-Lactate Dehydrogenase / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D014176:Protein Biosynthesis; D011506:Proteins / Q000187:drug effects*
D015415:Biomarkers; D006888:Hydroxycholesterols; D011506:Proteins; C007042:cholest-5-ene-3 beta,26-diol; C058194:26-aminocholesterol; C007997:25-hydroxycholesterol; D002784:Cholesterol; D007770:L-Lactate Dehydrogenase