A leukocyte type of 12-lipoxygenase is expressed in human vascular and mononuclear cells. Evidence for upregulation by angiotensin II.
The lipoxygenase (LO) pathway has been implicated in leading to accelerated atherosclerosis. However, the precise type of LO present in unstimulated human aortic smooth muscle cells (HSMC), endothelial cells (HAEC), and monocytes (MO) is not clear. In this study, we used a specific reverse-transcriptase polymerase chain reaction (RT-PCR) method to analyze the type of LO mRNA expressed in normal HSMC, HAEC, and MO. In all three cell types, a 333-base-pair band was seen when primers and probes specific for the leukocyte type of 12-LO were used, suggesting that a leukocyte type of 12-LO is expressed in these cell types. Western immunoblotting analysis in cultured HSMC, HAEC, and MO using a polyclonal peptide antibody to the leukocyte type of 12-LO showed a specific 72-kD band that is identical to the molecular weight of the leukocyte type of 12-LO. These results indicate that a leukocyte type of 12-LO RNA and protein are expressed in HSMC, HAEC, and MO. Further, angiotensin II upregulates 12-LO activity and expression in HSMC, supporting a role for this 12-LO pathway in human vascular disease.
Kim
J A
JA
City of Hope Medical Center, Duarte, CA 91010, USA.
Gu
J L
JL
Natarajan
R
R
Berliner
J A
JA
Nadler
J L
JL
D000804:Angiotensin II / Q000494:pharmacology*; D001011:Aorta / Q000201:enzymology; D001092:Arachidonate 12-Lipoxygenase / Q000235:genetics / Q000378:metabolism*; D001093:Arachidonate 15-Lipoxygenase / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000201:enzymology*; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D007962:Leukocytes / Q000201:enzymology*; D008969:Molecular Sequence Data; D009000:Monocytes / Q000201:enzymology*; D009131:Muscle, Smooth, Vascular / Q000201:enzymology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism
D012333:RNA, Messenger; D000804:Angiotensin II; D015847:Interleukin-4; D001092:Arachidonate 12-Lipoxygenase; D001093:Arachidonate 15-Lipoxygenase