The lipid peroxidation product 4-hydroxy-2-nonenal induces tissue factor decryption via ROS generation and the thioredoxin system.

Many pathophysiologic agents transform cryptic tissue factor (TF) on cells to prothrombotic TF, and one such stimulus is 4-hydroxy-2-nonenal (HNE), the most abundant aldehyde produced by the oxidation of ω-6 polyunsaturated fatty acids. HNE was shown to induce reactive oxygen species (ROS) generation and p38 MAPK activation, but the link between them and their role in TF decryption are unclear. The present study was carried out to elucidate potential mechanisms involved in HNE-induced TF decryption in monocytic cells. The data presented herein show that mitochondria are the primary source for HNE-induced ROS generation. The inhibition of mitochondrial electron transport chain complex III and V blocked HNE-induced ROS generation, but not p38 MAPK activation. These inhibitors reduced phosphatidylserine (PS) externalization and TF decryption significantly, but not completely. HNE treatment inhibited the activities of thioredoxin reductase (TrxR) and thioredoxin (Trx), independent of ROS. Inhibition of the TrxR/Trx system by HNE or pharmacological inhibitors induced p38 MAPK activation, PS externalization, and TF decryption. Additional studies revealed that the inhibition of TrxR/Trx led to activation of apoptosis signal-regulating kinase (ASK-1) and mitogen-activated protein kinase kinase 3/6. Inhibition of ASK-1 expression by small interfering RNA or its activity by pharmacological inhibitors diminished HNE-induced TF decryption. Overall, our data suggest that HNE induces TF decryption by 2 distinctive pathways. One is ROS dependent but independent of p38 MAPK activation, and the other is via TrxR/Trx and is p38 MAPK activation dependent. However, both mechanisms result in the enhancement of PS at the outer leaflet that is responsible for TF decryption.