Low concentration of oxidized low-density lipoprotein and lysophosphatidylcholine upregulate constitutive nitric oxide synthase mRNA expression in bovine aortic endothelial cells.
Endothelium-dependent relaxation is markedly reduced in atherosclerotic arteries. Recently, the endothelium-dependent relaxing factor has been identified as nitric oxide (NO). We used RNase protection assay and immunoblotting to elucidate the effect of atherogenic lipoprotein on the expression of constitutive NO synthase (cNOS) mRNA and protein levels in bovine aortic endothelial cells. Twenty-four-hour exposure to a low concentration of oxidized low-density lipoprotein (10 micrograms protein/mL) upregulated cNOS mRNA levels (2.4 +/- 0.4-fold, P < .01). However, native low-density lipoprotein and high-density lipoprotein did not have any effect on cNOS mRNA levels. Furthermore, 5 micrograms/mL of lysophosphatidylcholine (LPC) also upregulated cNOS mRNA levels (2.6 +/- 0.5-fold, P < .01) at 8 hours. This action of LPC was abolished with cycloheximide but not with staurosporine. We concluded that atherogenic lipoproteins upregulate cNOS mRNA and protein levels in bovine aortic endothelial cells. This observation supports the hypothesis that an impairment of endothelium-dependent vasodilatation in atherosclerotic vessels may not be due to a decrease in cNOS expression. Moreover, the LPC action on cNOS mRNA levels requires new protein synthesis.
Hirata
K
K
First Department of Internal Medicine, Kobe University School of Medicine, Japan.
Miki
N
N
Kuroda
Y
Y
Sakoda
T
T
Kawashima
S
S
Yokoyama
M
M
D000594:Amino Acid Oxidoreductases / Q000235:genetics*; D000818:Animals; D001011:Aorta; D002148:Calmodulin-Binding Proteins / Q000235:genetics*; D002417:Cattle; D002478:Cells, Cultured; D004730:Endothelium, Vascular / Q000166:cytology* / Q000378:metabolism; D015151:Immunoblotting; D008077:Lipoproteins, LDL / Q000032:analysis*; D008244:Lysophosphatidylcholines / Q000032:analysis*; D009252:NADPH Dehydrogenase / Q000235:genetics*; D019001:Nitric Oxide Synthase; D010084:Oxidation-Reduction; D015347:RNA Probes; D012333:RNA, Messenger / Q000032:analysis / Q000235:genetics*; D015854:Up-Regulation
D002148:Calmodulin-Binding Proteins; D008077:Lipoproteins, LDL; D008244:Lysophosphatidylcholines; D015347:RNA Probes; D012333:RNA, Messenger; D019001:Nitric Oxide Synthase; D000594:Amino Acid Oxidoreductases; D009252:NADPH Dehydrogenase