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Objective To investigate the effects of store-operated calcium entry (SOCE) on the bio-functions of vascular endothelial cells (ECs) based on human umbilical vein endothelial cells (HUVECs). Methods The cultured HUVECs were divided into control group and 2-APB (SOCE inhibitor, 10, 25, 50, 75 and 100 μmol/L) intervention groups. The SOCE was measured by the fluorescence intensity of calcium probe in 5 min after 2-APB treatment (n=10). The proliferation (n=9) and migration (n=10) capacities of HUVECs after different doses of 2-APB treatment (50, 75 and 100 μmol/L) for 24 h were measured by CCK-8 assay and Transwell assay. In addition, at 2 h after 2-APB treatment, Griess reagent kit was employed to detect the production of NO (n=6), and Western blotting was used to measure the expression of eNOS (n=3) in different groups. Results Low doses of 2-APB (10 and 20 μmol/L) showed an inhibitory effect on SOCE, but the inhibition had no statistical significance (P < 0.05), while high doses of 2-APB (50, 75 and 100 μmol/L) inhibited SOCE significantly (P>0.05). Treatment of 2-APB (50, 75 and 100 μmol/L) also suppressed the proliferation (decreased by 33.2%, 46.3% and 61.2%, respectively when compared with control cells, all P < 0.05), the migration (decreased by 33.5%, 54.3% and 64.9%, respectively when compared with control cells, all P < 0.05), NO production (decreased by 40.5%, 61.9% and 73.1%, respectively when compared with control cells, all P < 0.05) as well as the synthesis of eNOS (decreased by 45.1% and 62.4% in the 75 and 100 μmol/L treated cells, respectively when compared with control cells, both P < 0.05) in ECs. Conclusion SOCE is involved in the proliferation and migration of ECs, and inhibition of SOCE can induce ECs dysfunction, which may act through suppressing eNOS synthesis. |
