Kim J A, Gu J L, Natarajan R, Berliner J A, Nadler J L
City of Hope Medical Center, Duarte, CA 91010, USA.
Arterioscler Thromb Vasc Biol. 1995 Jul;15(7):942-8. doi: 10.1161/01.atv.15.7.942.
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.
脂氧合酶（LO）途径与动脉粥样硬化加速有关。然而，未受刺激的人主动脉平滑肌细胞（HSMC）、内皮细胞（HAEC）和单核细胞（MO）中存在的确切LO类型尚不清楚。在本研究中，我们使用特异性逆转录聚合酶链反应（RT-PCR）方法分析正常HSMC、HAEC和MO中表达的LO mRNA类型。在所有三种细胞类型中，当使用针对白细胞型12-LO的引物和探针时，可见一条333个碱基对的条带，表明这些细胞类型中表达白细胞型12-LO。使用针对白细胞型12-LO的多克隆肽抗体对培养的HSMC、HAEC和MO进行的Western免疫印迹分析显示，一条特异性的72-kD条带，其分子量与白细胞型12-LO相同。这些结果表明，白细胞型12-LO RNA和蛋白在HSMC、HAEC和MO中表达。此外，血管紧张素II上调HSMC中12-LO的活性和表达，支持该12-LO途径在人类血管疾病中的作用。
