Very low density lipoprotein and oxidized very low density lipoprotein induce monocyte chemotactic protein 1 in rabbit aortic smooth muscle cells.

OBJECTIVE

To understand whether very low density lipoprotein (VLDL) and oxidatively modified very low density lipoprotein (OX-VLDL) can stimulate vascular smooth muscle cells (SMCs) to express high level of monocyte chemotactic protein (MCP-1) mRNA.

METHODS

LDL and VLDL were isolated from normal blood donors by density gradient ultracentrifugation and oxidatively modified by adding CuCl2 and incubation at 37 degrees C for 24 hours. Rabbit aortic SMCs were cultured by a substrate-attached explant method, and their total RNA was extracted by guanidinium isothiocyanate method. The expression of MCP-1 mRNA in SMCs was examined by dot blotting analysis using a probe of gamma-22P-end-labeled 35-mer oligonucleotide.

RESULTS

Dot blotting analysis showed that cultured SMCs can express MCP-1 mRNA and the level of MCP-1 mRNA was increased significantly in SMCs after 24-hour coculture with VLDL (6-fold) and OX-VLDL (20-fold).

CONCLUSIONS

VLDL and OX-VLDL both can induce MCP-1 mRNA expression in cultured rabbit aortic SMCs and the latter is more effective than the former.