Altered sterol synthesis and its relationship to fluid-phase endocytosis in a macrophage cell line P388D1.
In a previous study glucocorticoids have been shown to depress the rate of fluid-phase endocytosis in a macrophage cell line, P388D1. This effect was observed when either fluorescein-labeled dextran or horseradish peroxidase (HRP) was used to measure endocytosis. In this report the relationship between cholesterol synthesis and endocytosis was examined in light of the ability of glucocorticoids to inhibit cholesterol biosynthesis. Two known inhibitors of cholesterol biosynthesis, ML-236B and 25-hydroxycholesterol (25-OH), were compared with dexamethasone (dex) for the ability to suppress endocytosis in cells grown in media supplemented with either 10% whole or delipidized neonatal bovine serum (NBS). In 10% whole serum all inhibitors reduced the uptake of HRP after 12 h incubation. Dexamethasone (1 microM) suppressed endocytosis by 30% whereas 25-OH (2.5 microM) and ML-236B (11.6 microM) inhibited by 38 and 52%, respectively. Supplementation of the growth medium with mevalonolactone (3.4 mM) prevented the inhibition of endocytosis by ML-236B. In contrast, mevalonolactone supplementation did not prevent either dex or 25-OH from suppressing endocytosis. The same pattern of results was obtained when cultures were grown in delipidized NBS. After 4 h all inhibitors caused a decrease in amount of [14C]acetate incorporated into both nonsaponifiable lipids and digitonin precipitable sterols. Although dex inhibited cholesterol biosynthesis, total cellular cholesterol was unaffected by dex treatment after 24 h incubation. It is suggested that in addition to suppressing mevalonate synthesis, 25-OH, and by analogy dex, may act at some metabolic site(s) distal to the formation of mevalonate.
D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology; D002460:Cell Line; D002478:Cells, Cultured; D002784:Cholesterol / Q000096:biosynthesis; D004705:Endocytosis / Q000187:drug effects; D008264:Macrophages / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D013261:Sterols / Q000037:antagonists & inhibitors / Q000096:biosynthesis*; D013997:Time Factors
D000924:Anticholesteremic Agents; D013261:Sterols; D002784:Cholesterol