TY - JOUR
T1 - Activation of cytosolic phospholipase A2 and 15-lipoxygenase by oxidized low-density lipoproteins in cultured human lung fibroblasts
AU - Lupo, Gabriella
AU - Anfuso, Carmelina Daniela
AU - Ragusa, Nicolò
AU - Tirolo, Cataldo
AU - Marchetti, Bianca
AU - Gili, Elisa
AU - Rosa, Cristina La
AU - Vancheri, Carlo
PY - 2007/4
Y1 - 2007/4
N2 - In cell cultures of human lung fibroblasts, we found that oxidized LDL (oxLDL), after 24-h treatment, stimulated arachidonic acid release. A putative role for phospholipases A2 and MAPK activities in this process was postulated. Consequently, we studied the contribution of either Ca2+-dependent, cytosolic phospholipase A2 (cPLA2) or Ca2+-independent phospholipase A2 (iPLA2), and the role of the MAP kinase family in oxLDL toxicity to fibroblastic cells in vitro. Activation of extracellular signal-regulated kinases ERK1/2, p38 and c-Jun NH2-terminal kinase (JNK) was also assessed with Western blotting. Compared with cellular samples untreated or treated with native LDL, treatment with oxLDL (50-100 μM hydroperoxides) for 24 h significantly increased the levels of either cPLA2 protein expression or constitutively phosphorylated cPLA2 protein; in addition we observed enzyme translocation to membranes. iPLA2 activity was not stimulated by oxLDL. Arachidonic acid release appeared to be associated with phosphorylation of ERK1/2 which was significantly enhanced in a dose-dependent manner whereas no activation of p38 and JNKs was found, indicating that these MAPKs are not involved in mediating the maximal oxLDL response. Western blotting on subcellular fractions and confocal microscopy analyses confirmed an increase in 15-lipoxygenase (15-LO) protein expression and translocation upon activation. A significant increase of cyclooxygenase-2 expression into membrane fraction was also found. Collectively, the data presented link the stimulation of ERK-cPLA2-15-LO pathway by oxLDL to the prooxidant mechanism of the lipoprotein complex. It may initially stimulate the fibroblast reaction against the oxidation challenge as well as metabolic repair, such as during lung inflammation and pulmonary fibrosis.
AB - In cell cultures of human lung fibroblasts, we found that oxidized LDL (oxLDL), after 24-h treatment, stimulated arachidonic acid release. A putative role for phospholipases A2 and MAPK activities in this process was postulated. Consequently, we studied the contribution of either Ca2+-dependent, cytosolic phospholipase A2 (cPLA2) or Ca2+-independent phospholipase A2 (iPLA2), and the role of the MAP kinase family in oxLDL toxicity to fibroblastic cells in vitro. Activation of extracellular signal-regulated kinases ERK1/2, p38 and c-Jun NH2-terminal kinase (JNK) was also assessed with Western blotting. Compared with cellular samples untreated or treated with native LDL, treatment with oxLDL (50-100 μM hydroperoxides) for 24 h significantly increased the levels of either cPLA2 protein expression or constitutively phosphorylated cPLA2 protein; in addition we observed enzyme translocation to membranes. iPLA2 activity was not stimulated by oxLDL. Arachidonic acid release appeared to be associated with phosphorylation of ERK1/2 which was significantly enhanced in a dose-dependent manner whereas no activation of p38 and JNKs was found, indicating that these MAPKs are not involved in mediating the maximal oxLDL response. Western blotting on subcellular fractions and confocal microscopy analyses confirmed an increase in 15-lipoxygenase (15-LO) protein expression and translocation upon activation. A significant increase of cyclooxygenase-2 expression into membrane fraction was also found. Collectively, the data presented link the stimulation of ERK-cPLA2-15-LO pathway by oxLDL to the prooxidant mechanism of the lipoprotein complex. It may initially stimulate the fibroblast reaction against the oxidation challenge as well as metabolic repair, such as during lung inflammation and pulmonary fibrosis.
KW - 15-lipoxygenase
KW - Fibroblasts
KW - Low-density lipoprotein
KW - MAP kinases
KW - Phospholipase A
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U2 - 10.1016/j.bbalip.2007.01.014
DO - 10.1016/j.bbalip.2007.01.014
M3 - Article
C2 - 17344094
AN - SCOPUS:34047150002
VL - 1771
SP - 522
EP - 532
JO - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
JF - Biochimica et Biophysica Acta - Molecular and Cell Biology of Lipids
SN - 1388-1981
IS - 4
ER -