Phosphorylation of 5-lipoxygenase at ser523 by protein kinase A determines whether pioglitazone and atorvastatin induce proinflammatory leukotriene B4 or anti-inflammatory 15-epi-lipoxin a4 production.
The 5-lipoxygenase (5LO) produces leukotriene B(4) and 15-epilipoxin-A(4) (15-epi-LXA(4)). Phosphorylation at Ser(523) by protein kinase A (PKA) prevents 5LO shift to the perinuclear membrane. Atorvastatin and pioglitazone up-regulate 15-epi-LXA(4) production in the heart. We assessed whether phosphorylation of 5LO by PKA determines whether 5LO interacts with the membranous cytosolic phospholipase A(2) (cPLA(2)) to produce leukotriene B(4) or with cyclooxygenase-2 (COX2) to produce 15-epi-LXA(4). Rats received either pioglitazone, atorvastatin, pioglitazone plus atorvastatin, vehicle, or LPS. Rat myocardial cells were incubated with pioglitazone plus atorvastatin, pioglitazone plus atorvastatin plus H-89 (PKA inhibitor), H-89, or vehicle for 8 h. Pioglitazone and atorvastatin did not affect total 5LO expression. However, both increased 5LO levels in the cytosolic fraction. H-89 caused a shift of 5LO to the membranous fraction in atorvastatin- and pioglitazone-treated rats. Pioglitazone and atorvastatin increased phospho-5LO levels. H-89 attenuated this increase. Both pioglitazone and atorvastatin increased COX2 levels in the cytosolic fraction and the membranous fraction. H-89 prevented this increase. Pioglitazone and atorvastatin increased cPLA(2) expression in the membranous fraction. This effect was not attenuated by H-89. Pioglitazone plus atorvastatin increased 15-epi-LXA(4) levels. H-89 attenuated the effect of pioglitazone plus atorvastatin. Pioglitazone plus atorvastatin plus H-89 increased leukotriene B(4) levels. Coimmunoprecipitation showed that without H-89, atorvastatin and pioglitazone induced an interaction between 5LO and COX2 in the cytosolic fraction, whereas when H-89 was added, 5LO interacted with cPLA(2) on the membranous fraction. The 5LO phosphorylation determines whether 15-epi-LXA(4) (anti-inflammatory) or leukotriene B(4) (inflammatory mediator) is produced.
Ye
Yumei
Y
Division of Cardiology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Lin
Yu
Y
Perez-Polo
Jose R
JR
Uretsky
Barry F
BF
Ye
Zaiming
Z
Tieu
Brian C
BC
Birnbaum
Yochai
Y
D000818:Animals; D001094:Arachidonate 5-Lipoxygenase / Q000378:metabolism*; D000069059:Atorvastatin; D017868:Cyclic AMP-Dependent Protein Kinases / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000187:drug effects; D006538:Heptanoic Acids / Q000008:administration & dosage / Q000494:pharmacology*; D007155:Immunologic Factors / Q000235:genetics; D007546:Isoquinolines / Q000494:pharmacology; D007975:Leukotriene B4 / Q000096:biosynthesis*; D044045:Lipoxins / Q000096:biosynthesis*; D008297:Male; D009206:Myocardium / Q000166:cytology / Q000276:immunology; D010766:Phosphorylation; D000077205:Pioglitazone; D011758:Pyrroles / Q000008:administration & dosage / Q000494:pharmacology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012694:Serine / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D045162:Thiazolidinediones / Q000008:administration & dosage / Q000494:pharmacology*
D006538:Heptanoic Acids; D007155:Immunologic Factors; D007546:Isoquinolines; D044045:Lipoxins; D011758:Pyrroles; D013449:Sulfonamides; D045162:Thiazolidinediones; C040527:lipoxin A4; D007975:Leukotriene B4; D012694:Serine; D000069059:Atorvastatin; D001094:Arachidonate 5-Lipoxygenase; D017868:Cyclic AMP-Dependent Protein Kinases; C063509:N-(2-(4-bromocinnamylamino)ethyl)-5-isoquinolinesulfonamide; D000077205:Pioglitazone