25-hydroxycholesterol increases eicosanoids and alters morphology in cultured pulmonary artery smooth muscle and endothelial cells.
25-hydroxycholesterol (25-OHC) is an oxidized derivative of cholesterol that has been implicated in the early development of arteriosclerosis. Changes in arterial smooth muscle cell (SMC) migration and proliferation have also been linked to the pathophysiology of arteriosclerosis. SMCs undergo "activation" in response to vascular injury by changing phenotypically and by increasing prostaglandin G/H synthase-2 (PGHS-2) protein levels and eicosanoid release. Activation is thought to be important in atheroma formation and arteriosclerosis progression. 25-OHC induces SMCs to change morphologically, increase PGHS-2, and increase eicosanoid release. Confluent monolayers were treated with 25-OHC (10 microg/mL) or the PGHS-2 inducer interleukin-1beta (1 ng/mL) for 18 hours at 37 degrees C. The 18-hour treatment resulted in morphological changes. After uptake of [(14)C]arachidonic acid, released radiolabeled arachidonic acid products were extracted and chromatographed by both normal and reverse-phase high-performance liquid chromatography systems. 25-OHC-treated cells increased their prostaglandin production, with the major component comigrating with a prostaglandin-E(2) standard. HETEs and epoxyeicosatrienoic acids were not affected. Immunoprecipitation analysis of treated and control cell lysates using anti-PGHS-1 and -2 and anti-alpha-actin primary antibodies indicated PGHS-2 induction over control and no change in contractile proteins. These changes are consistent with SMC activation, which occurs in vascular injury models. The notion that oxysterols can activate vascular SMCs may be important in ultimately understanding the pathophysiology of atheroma formation.
Wohlfeil
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ER
Department of Anesthesiology, Medical College of Wisconsin, Milwaukee 53226, USA. wohlfeil@mcw.edu
Campbell
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WB
D000199:Actins / Q000032:analysis / Q000378:metabolism; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism / Q000494:pharmacology; D002250:Carbon Radioisotopes; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D051546:Cyclooxygenase 2; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D006888:Hydroxycholesterols / Q000494:pharmacology*; D006893:Hydroxyeicosatetraenoic Acids / Q000032:analysis / Q000096:biosynthesis; D007527:Isoenzymes / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects / Q000201:enzymology*; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D011453:Prostaglandins / Q000032:analysis / Q000096:biosynthesis*; D011651:Pulmonary Artery / Q000166:cytology; D011817:Rabbits; D013347:Subcellular Fractions / Q000737:chemistry / Q000378:metabolism
D000199:Actins; D002250:Carbon Radioisotopes; D006888:Hydroxycholesterols; D006893:Hydroxyeicosatetraenoic Acids; D007527:Isoenzymes; D011453:Prostaglandins; D016718:Arachidonic Acid; C027356:11-hydroxy-5,8,12,14-eicosatetraenoic acid; C025984:15-hydroxy-5,8,11,13-eicosatetraenoic acid; C007997:25-hydroxycholesterol; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases