triolein and Wolman-Disease

High-density lipoprotein (HDL)-[3H]triolein (i.e. [3H]triolein incorporated into reconstituted HDL) was taken up by cultured fibroblasts through an apparently saturable process, competitively inhibited by non-labelled HDL and independent of the LDL receptor. Using 125I-HDL and HDL-[3H]triolein, binding experiments (at 0 degrees C) followed by a short-time 'chase' at 37 degrees C showed that 125I radioactivity was rapidly released in the culture medium (as trichloroacetic acid-precipitable material), whereas 3H radioactivity remained associated with the cell. The cell-associated HDL-[3H]triolein was rapidly degraded in normal fibroblasts, and the liberated [3H]oleic acid was incorporated into newly biosynthesized phospholipids. In Wolman-disease fibroblasts HDL-[3H]triolein was degraded at a normal rate, and thus independently of the lysosomal compartment. In contrast, the degradation of HDL-[3H]triolein was blocked in fibroblasts from Neutral Lipid Storage Disease (NLSD), similarly to that of endogenously biosynthesized triacylglycerols [Radom, Salvayre, Nègre, Maret and Douste-Blazy (1987) Eur. J. Biochem. 164, 703-708]. Trypsin-treated HDL-[3H]triolein was also taken up by cells and degraded quite similarly to HDL-[3H]triolein. In conclusion, all these data taken together suggest that HDL-[3H]triolein is: (i) associated with the cell through a process independent of intact apolipoprotein (apo) As, thus probably independent of an apoA-receptor-mediated uptake; (ii) internalized by cells, whereas 125I-apoAs are released in the culture medium; (iii) directed to the same non-lysosomal catabolic pool (blocked in NLSD) as for endogenously biosynthesized triacylglycerols.

Topics: Cells, Cultured; Fibroblasts; Humans; Lipid Metabolism, Inborn Errors; Lipoproteins, HDL; Lipoproteins, LDL; Triglycerides; Triolein; Wolman Disease

The functional relationship between the two subcellular compartments involved in catabolism of triglycerides, i.e. lysosomes and lipid-containing cytoplasmic vacuoles, has been investigated using cultured fibroblasts from patients affected with two different genetic lipid (triacylglycerol) storage disorders: Wolman disease and multisystemic lipid storage myopathy. As shown by metabolic studies in intact cultured cells, lysosomal degradation of exogenous labelled triacylglycerols (incorporated into lipoproteins and internalized via the apo B/E receptor pathway) was blocked in Wolman cells, whereas catabolism of endogenously biosynthesized triacylglycerols was in the normal range. In contrast, in fibroblasts from multisystemic lipid storage myopathy, the degradation of endogenous triacylglycerols was blocked, whereas that of exogenous triacylglycerols (i.e. from lipoproteins) was normal. This comparative study demonstrates that the lysosomal and cytoplasmic compartments are functionally independent. Enzymatic studies allows one to discriminate clearly between 3 lipases and 2 carboxylesterases the role of which is discussed.

Topics: Cells, Cultured; Fibroblasts; Humans; Hydrogen-Ion Concentration; Kinetics; Lipid Metabolism, Inborn Errors; Muscular Diseases; Skin; Triglycerides; Triolein; Wolman Disease