[1]王连友,杨杰,黄岚.钙库操纵性钙内流参与调节血管内皮细胞增殖与迁移[J].第三军医大学学报,2019,41(05):424-429.
 WANG Lianyou,YANG Jie,HUANG Lan.Store-operated calcium entry is involved in proliferation and migration of vascular endothelial cells[J].J Third Mil Med Univ,2019,41(05):424-429.
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钙库操纵性钙内流参与调节血管内皮细胞增殖与迁移(/HTML )
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《第三军医大学学报》[ISSN:1000-5404/CN:51-1095/R]

卷:
41卷
期数:
2019年第05期
页码:
424-429
栏目:
基础医学
出版日期:
2019-03-15

文章信息/Info

Title:
Store-operated calcium entry is involved in proliferation and migration of vascular endothelial cells
作者:
王连友杨杰黄岚
解放军第988医院心内科;陆军军医大学(第三军医大学)第二附属医院心内科
Author(s):
WANG Lianyou YANG Jie HUANG Lan

Department of Cardiology, No. 988 Hospital of PLA, Zhengzhou, Henan Province, 450042; Department of Cardiology, Second Affiliated Hospital, Army Medical University (Third Military Medical University), Chongqing, 400037, China

关键词:
血管内皮细胞内皮细胞功能异常增殖迁移钙库操纵性钙内流
Keywords:
vascular endothelial cells endothelial cells dysfunctionproliferationmigrationstore-operated calcium entry
分类号:
R322.12; R329.28; R348.2
文献标志码:
A
摘要:

目的 利用体外培养的人脐静脉血管内皮细胞,初步探讨钙库操纵性钙内流(store-operated calcium entry,SOCE)对血管内皮细胞(endothelial cell,EC)功能的影响。方法 培养于激光共聚焦培养皿的EC随机分为对照组和不同剂量(10、25、50、75、100 μmol/L)SOCE抑制剂(2-APB)干预组。干预组细胞以2-APB干预5 min后,检测各组细胞的SOCE强度(n=10)。培养于6孔板的细胞,根据实验目的,随机分为对照组和不同剂量(50、75、100 μmol/L)的2-APB干预组。2-APB干预24 h后,以CCK-8检测各组EC的增殖功能(n=9)、以Transwell培养法检测各组EC的迁移功能(n=10)。2-APB干预2 h后,以Griess Reagent方法检测各组EC的NO产量(n=6)以及用Western blot方法检测各组EC的eNOS变化(n=3)。结果 较低剂量的2-APB(10、25 μmol/L)对SOCE有抑制趋势,但无统计学意义(P>0.05),较高剂量的2-APB(50、75、100 μmol/L)能够显著抑制SOCE(P<0.05)。采用具有显著抑制作用浓度的2-APB(50、75、100 μmol/L)抑制SOCE后,能够显著抑制EC的增殖能力(与对照组比较,EC的增殖分别下降33.2%、46.3%和61.2%,P<0.05)、迁移功能(与对照组比较,分别下降33.5%、54.3%和64.9%,P<0.05)、NO产量(与对照组比较,分别下降40.5%、61.9%和73.1%,P<0.05)以及eNOS的合成(与对照组比较,75和100 μmol/L组分别下降45.1%和62.4%,P<0.05)。结论 SOCE参与调节EC的增殖和迁移功能,抑制SOCE能引起EC功能异常,此作用可能是通过抑制eNOS的合成实现。

Abstract:

ObjectiveTo 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.

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更新日期/Last Update: 2019-03-05