COX-1 and COX-2 contribute differentially to the LPS-induced release of PGE2 and TxA2 in liver macrophages.

LPS induces an immediate release of thromboxane TxA2 and a delayed release of PGE2. Dexamethasone suppresses the LPS-induced release of TxA2 and PGE2. In the first 8 h after LPS addition, the specific COX-2 inhibitor SC236 inhibits the PGE2 and TxA2 release by about 80% and 20%, whereas the release of PGE2 and TxA2 between 8 and 24 h is inhibited by about 40% and 35%, respectively. Resident liver macrophages express substantial amounts of COX-1, TxAS, cPGES and mPGES-2, small amounts of COX-2 but almost no detectable amounts of mPGES-1. LPS induces an increase of COX-2 and mPGES-1, but does not change COX-1, cPGES, mPGES-2 and TxAS at protein level. Dexamethasone suppresses almost completely the LPS-induced effects on COX-2 and mPGES-1. It is concluded that (1) COX-1 and COX-2 are involved in the LPS-induced synthesis of TxA2 and PGE2; (2) TxA2 release is catalyzed at early time-points by the combined action of COX-1 and TxAs, whereas at later time points the newly expressed COX-2 couples to TxAS and contributes to the TxA2 release; (3) PGE2 release within the first 8 h is predominantly catalyzed by COX-2, whereas at later time-points COX-1 couples to the newly expressed mPGES-1 and contributes to the PGE2 release.

Diverse functional coupling of cyclooxygenase 1 and 2 with final prostanoid synthases in liver macrophages.

Lipopolysaccharide (LPS) treatment of resident liver macrophages resulted in a coordinated enhanced expression of cytosolic phospholipase A(2) (cPLA(2)), cyclooxygenase (COX)-2 and prostaglandin E(2)-synthase. LPS-pretreated liver macrophages showed a higher release of PGE(2) after zymosan, phorbol ester and A23187, of PGF(2alpha) after zymosan and A23187, whereas the release of thromboxane B(2) and PGD(2) was unchanged. Inhibition of COX-1 and -2 by specific inhibitors (SC560, SC236) inhibited the prostanoid release between 50-80% and 20-40%, respectively, indicating a predominant role for COX-1. In detail (1) the zymosan-induced release of all prostanoids was inhibited to a similar degree by the COX-1 inhibitor (about 70%) and the COX-2 inhibitor (20-30%), (2) PGE(2) release after all stimuli was inhibited to a greater extent by SC560 (70-90%) compared to SC236 (5-30%), (3) the phorbol ester- and A23187-induced release of PGF(2alpha) and PGD(2) was inhibited equally (40-50%) by both inhibitors, (3) TxB(2) release after phorbol ester and A23187 was inhibited by SC560 by 50 and 30%, and by SC236 by 50 and 70%, respectively. cPLA(2), COX-1 and -2, and the final prostanoid synthases were found in different subcellular fractions. These results indicate, that the functional coupling of COX-1 and -2 to final prostanoid synthases depends on the stimulation of the cells.

Lipopolysaccharide-induced release of arachidonic acid and prostaglandins in liver macrophages: regulation by Group IV cytosolic phospholipase A2, but not by Group V and Group IIA secretory phospholipase A2.

Lipopolysaccharide (LPS) induces a delayed release (lag phase of 2-4 h) of arachidonic acid (AA) and prostaglandin (PG) D2 in rat liver macrophages. Group IV cytosolic phospholipase A2 (cPLA2) becomes phosphorylated within minutes after the addition of LPS. The phosphorylated form of cPLA2 shows an enhanced in vitro activity. The Ca2+ dependence of cPLA2 activity is not affected by phosphorylation of the enzyme. In addition, LPS induces an enhanced expression of cPLA2 mRNA (after 2-4 h) and an enhanced expression of cPLA2 protein (after 8 h). The cellular cPLA2 activity is enhanced about twofold 24 h after LPS treatment. Liver macrophages constitutively express mRNAs encoding Groups V and IIA secretory PLA2 (sPLA2). LPS has no effect on the levels of Groups V and IIA sPLA2 mRNA expression. Despite mRNA expression, Groups V and IIA sPLA2 protein and sPLA2 activity are not detectable in unstimulated or LPS-stimulated liver macrophages. Collectively, these and earlier [Mediators Inflammation 8 (1999) 295.] results suggest that in liver macrophages the LPS-induced delayed release of AA and prostanoids is mediated by phosphorylation and an enhanced expression of cPLA2, a de novo expression of cyclooxygenase (COX)-2, but not by the actions of Group V or Group IIA sPLA2.