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Microvesicles derived from hypoxia/reoxygenation-treated human umbilical vein endothelial cells impair relaxation of rat thoracic aortic rings / 中国应用生理学杂志
Chinese Journal of Applied Physiology ; (6): 560-566, 2014.
Article in English | WPRIM | ID: wpr-243411
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effects of microvesicles (MVs) derived from hypoxia/reoxygenation (H/R)-treated human umbilical vein endothelial cells (HUVECs) on endothelium-dependent relaxation of rat thoracic aortic rings.</p><p><b>METHODS</b>H/R injury model was established to induce HUVECs to release H/R-EMVs. H/R-EMVs from HUVECs were isolated by ultracentrifugation from the conditioned culture medium. H/R-EMVs were characterized using 1 μm latex beads and anti-PE-CD144 by flow cytometry. Thoracic aortic rings of rats were incubated with 2.5, 5, 10, 20 μg/ml H/R-EMVs derived from H/R-treated HUVECs for 4 hours, and their endothelium-dependent relaxation in response to acetylcholine (ACh) or endothelium-independent relaxation in response to sodium nitroprusside (SNP) was recorded in vitro. The nitric oxide (NO) production of ACh-treated thoracic aortic rings of rats was measured using Griess reagent. The expression of endothelial NO synthase (eNOS) and phosphorylated eNOS (p-eNOS, Ser-1177) in the thoracic aortic rings of rats was detected by Western blotting. Furthermore, the levels of SOD and MDA in H/R-EMVs-treated thoracic aortic rings of rats were measured using SOD and MDA kit.</p><p><b>RESULTS</b>H/R-EMVs were induced by H/R-treated HUVECs and isolated by ultracentrifugation. The membrane vesicles (< 1 μm) induced by H/R were CD144 positive. ACh-induced relaxation and NO production of rat thoracic aortic rings were impaired by H/R-EMVs treatment in a concentration-dependent manner (P < 0.05, P < 0.01). The expression of total eNOS (t-eNOS) was not affected by H/R-EMVs. However, the expression of p-eNOS decreased after treated with H/R-EMVs. The activity of SOD decreased and the level of MDA increased in H/R-EMVs treated rat thoracic aortic rings (P < 0.01).</p><p><b>CONCLUSION</b>ACh induced endothelium-dependent relaxation of thoracic aortic rings of rats was impaired by H/R-EMVs in a concentration-dependent manner. The mechanisms included a decrease in NO production, p-eNOS expression and an increase in oxidative stress.</p>
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Aorta, Thoracic / Pharmacology / Physiology / In Vitro Techniques / Nitroprusside / Endothelium, Vascular / Acetylcholine / Cell Hypoxia / Oxidative Stress / Cell Biology Limits: Animals / Humans Language: English Journal: Chinese Journal of Applied Physiology Year: 2014 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Aorta, Thoracic / Pharmacology / Physiology / In Vitro Techniques / Nitroprusside / Endothelium, Vascular / Acetylcholine / Cell Hypoxia / Oxidative Stress / Cell Biology Limits: Animals / Humans Language: English Journal: Chinese Journal of Applied Physiology Year: 2014 Type: Article