Fenofibrate modifies human vascular smooth muscle proteoglycans and reduces lipoprotein binding.
OBJECTIVE
Vascular disease in type 2 diabetes is associated with an up-regulation of atherogenic growth factors, which stimulate matrix synthesis including proteoglycans. We have examined the direct actions of fenofibrate on human vascular smooth muscle cells (VSMCs) and have specifically investigated proteoglycan synthesis and binding to LDL.


METHODS
Proteoglycans synthesised by human VSMCs treated with fenofibrate (30 micromol/l) were assessed for binding to human LDL using a gel mobility shift assay, metabolically labelled with [(35)S]-sulphate and quantitated by cetylpyridinium chloride. They were then assessed for electrophoretic mobility by SDS-PAGE, for size by gel filtration, for sulphation pattern by fluorophore-assisted carbohydrate electrophoresis, and for glycosaminoglycan (GAG) composition by enzyme digestion.


RESULTS
Proteoglycans synthesised in the presence of fenofibrate showed an increase in the half-maximum saturation concentration of LDL from 36.8+/-12.4 microg/ml to 77.7+/-17 microg/ml under basal conditions, from 24.9+/-4.6 microg/ml to 39.1+/-6.1 microg/ml in the presence of TGF-beta1, and from 9.5+/-4.4 microg/ml to 31.1+/-3.4 microg/ml in the presence of platelet-derived growth factor/insulin. Fenofibrate treatment in the presence of TGF-beta1 inhibited the incorporation of [(35)S]-sulphate into secreted and cell-associated proteoglycans synthesised by human VSMCs by 59.2% (p<0.01) and 39.8% (p<0.01) respectively. The changes in sulphate incorporation following treatment with fenofibrate were associated with a concentration-related increase in the electrophoretic mobility due to a reduction in GAG length. There was no change in the sulphation pattern; however, there was an alteration in the disaccharide composition of the GAGs.


CONCLUSIONS
Fenofibrate modifies the structure of vascular proteoglycans by reducing the length of the GAG chains and GAG composition, resulting in reduced binding to human LDL, a mechanism which may lead to a reduction of atherosclerosis and cardiovascular disease in people with diabetes treated with fenofibrate.
Nigro
J
J
Cell Biology of Diabetes Laboratory, Baker Heart Research Institute, St. Kilda Rd Central, PO Box 6492, Melbourne, Victoria, 8008, Australia.
Ballinger
M L
ML
Dilley
R J
RJ
Jennings
G L R
GL
Wight
T N
TN
Little
P J
PJ
D002478:Cells, Cultured; D011345:Fenofibrate / Q000494:pharmacology*; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D000960:Hypolipidemic Agents / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology; D008074:Lipoproteins / Q000187:drug effects / Q000378:metabolism*; D008323:Mammary Arteries; D009131:Muscle, Smooth, Vascular / Q000187:drug effects / Q000502:physiology*; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D011509:Proteoglycans / Q000187:drug effects / Q000378:metabolism*; D016212:Transforming Growth Factor beta / Q000494:pharmacology
D006025:Glycosaminoglycans; D000960:Hypolipidemic Agents; D007328:Insulin; D008074:Lipoproteins; D010982:Platelet-Derived Growth Factor; D011509:Proteoglycans; D016212:Transforming Growth Factor beta; D011345:Fenofibrate