Identification of cytochrome P450s involved in the metabolism of arachidonic acid in human platelets.
Although cytochrome P450s (CYPs) have been identified in most human cells, identification of CYPs in human platelets remains poorly explored. CYP expressions in human platelets were screened by using reverse transcriptase-polymerase chain reaction and western blot analysis followed by functional assays using arachidonic acid (ARA). CYP1A1, 2U1, 2J2, 4A11, 4F2, and 5A1 were expressed as both proteins and mRNAs in platelets. Ethoxyresorufin-O-deethylase activity was observed in platelets and this activity was significantly decreased after treatment with the general P450 inhibitor SKF-525A and the CYP1A inhibitor, α-naphthoflavone (40-45%, P<0.001). Seventeen ARA metabolites were detected in ARA-treated platelets. Among these, the levels of 20-hydroxyeicosatetraenoic acid and epoxyeicosatrienoic acids were significantly decreased with the treatment of the P450 ω-hydroxylase inhibitor 17-octadecynoic acid (P<0.05-0.001). In summary, multiple ARA-metabolizing P450s were identified in human platelets. These findings may provide an important resource for understanding physiological function of platelet.
Department of Pharmacology, Pharmacogenomics Research Center, Inje University College of Medicine, Inje University, Busan, South Korea.
Yazun B
YB
Jarrar
Sun-Ah
SA
Cho
Kyung-Suk
KS
Oh
Dong-Hyun
DH
Kim
Jae-Gook
JG
Shin
Su-Jun
SJ
Lee
D000328:Adult; D001095:Arachidonic Acids / Q000378:metabolism*; D001792:Blood Platelets / Q000201:enzymology*; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics / Q000378:metabolism*; D005231:Fatty Acids, Unsaturated / Q000494:pharmacology; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction; D059467:Transcriptome
D001095:Arachidonic Acids; D065607:Cytochrome P-450 Enzyme Inhibitors; D005231:Fatty Acids, Unsaturated; C047008:17-octadecynoic acid; D003577:Cytochrome P-450 Enzyme System
Arachidonic acid; Cytochrome P450; Metabolism; Platelet