Regulation of Mitogen-Activated Protein Kinase Cascades by Low Density Lipoprotein and Lysophosphatidic Acid

Aim of the present study was to elucidate the regulation of the mitogen activated protein (MAP) kinase cascades as well as the cross-talk between the various MAP kinase isoforms (ERK1/2, SAPK and p38) after stimulation of vascular smooth muscle cells (VSMC) with low density lipoprotein (LDL), 100 µg/ml or lysophosphatidic acid (LPA), 5 µg/ml. Furthermore, the role of the intracellular free Ca 2+ concentration ([Ca 2+ ] i ) on the activation of the MAP kinase isoforms as well as on the protein expression of MAP kinase phosphatase (MKP)-1 was investigated. The methods used were Western blot analysis, immunoprecipitation and LDL isolation. Involvement of G i -proteins on LDL- and LPA-induced activation of the MAP kinase isoforms was examined by treatment of VSMC with pertussis toxin (PTX),100 ng/ml. LDL as well as LPA induced an activation of SAPK and p38 MAP kinase in a PTX-sensitive manner. The ERK1/2, SAPK and p38 MAP kinase activation was a Ca 2+ -dependent process, most likely regulated through modulation of MKP-1 protein expression. Inhibition of ERK1/2 by PD 98059 completely abolished LDL- and LPA-induced activation of SAPK, whereas the activation of p38 MAP kinase was not affected. We conclude, that [Ca 2+ ] i regulates the PTX-sensitive LDL- and LPA-induced stimulation of the MAP kinase cascades, probably via inhibition of the MKP-1 protein expression.