Store-operated calcium entry involves in regulating the hyperpermeability of endothelial cells induced by ox-LDL

Yue KANG; Suyu TAI; Maowei LI; Linjun YU; Lianyou WANG

doi:10.16016/j.2097-0927.202207164

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Store-operated calcium entry involves in regulating the hyperpermeability of endothelial cells induced by ox-LDL

Basic Medicine | 更新时间：2026-01-09

- Store-operated calcium entry involves in regulating the hyperpermeability of endothelial cells induced by ox-LDL
- Journal of Army Medical University Vol. 45, Issue 4, Pages: 358-365(2023)
- 作者机构：
  
  1.453003 河南新乡，新乡医学院研究生院
  2.450042 郑州，联勤保障部队988医院心内科
- 作者简介：
- 基金信息：
  
  the Henan Province-Ministry Co-Construct Key Medical Science and Technology Project(LHGJ20190865)
- DOI：10.16016/j.2097-0927.202207164
  CLC： R322.123;R329.27;R363.21
- Received：25 July 2022，
  
  Revised：2022-09-29，
  
  Published：28 February 2023
- 稿件说明：
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Yue KANG, Suyu TAI, Maowei LI, et al. Store-operated calcium entry involves in regulating the hyperpermeability of endothelial cells induced by ox-LDL[J]. Journal of Army Medical University, 2023, 45(4): 358-365.
DOI：

Yue KANG, Suyu TAI, Maowei LI, et al. Store-operated calcium entry involves in regulating the hyperpermeability of endothelial cells induced by ox-LDL[J]. Journal of Army Medical University, 2023, 45(4): 358-365. DOI： 10.16016/j.2097-0927.202207164.

摘要

目的

利用体外培养的人脐静脉血管内皮细胞(human umbilical vein endothelial cells，HUVECs)，探讨钙库操纵性钙内流(store-operated calcium entry，SOCE)在氧化低密度脂蛋白(oxidized low-density lipoprotein，ox-LDL)诱导的血管内皮细胞通透性改变中的作用。

方法

体外培养HUVECs，传代3次后，用于检测跨内皮电阻(transendothelial electrical resistance，TER)的细胞随机分为对照组和不同剂量ox-LDL(10、25、50、100 μg/mL)干预组(

=3)。培养于载玻片上的细胞随机分为对照组和ox-LDL 24 h组、ox-LDL 48 h组，免疫荧光检测细胞骨架蛋白F-actin及细胞连接蛋白VE-cadherin的分布，免疫印迹检测VE-cadherin的含量。细胞孵育钙离子探针(Fluo-3/AM)后，激光共聚焦检测细胞内钙含量(对照组、ox-LDL组、ox-LDL+2APB组)以及SOCE(对照组、ox-LDL 24h组、ox-LDL 48 h组)。

结果

低剂量的ox-LDL(10、25 μg/ mL)对TER无明显影响，高剂量ox-LDL(50、100 μg/ mL)在6 h内对TER影响不明显，但在12 h引起TER降低(

0.05)，此效应在24 h和48 h更加明显(

0.05)。Ox-LDL(100 μg/mL)处理后24、48 h，VE-cadherin的表达减少(

0.05)，并影响F-actin的形态和分布；HUVECs的游离钙浓度显著增加(

0.05)，此效应被SOCE抑制剂2-APB阻断，2-APB能够减弱ox-LDL对TER的影响。ox-LDL(100 μg/mL)处理48 h后，SOCE的振幅显著降低(

0.05)。SOCE降支速度在ox-LDL处理24 h后呈现降低趋势，而在ox-LDL处理48 h后呈显著下降(

0.05)。

结论

ox-LDL通过延长内皮细胞SOCE的持续时间上调细胞内游离钙浓度，影响细胞骨架的排列和细胞间连接的完整性，引起细胞通透性的增加。

Abstract

Objective

To explore the effects of store-operated calcium entry(SOCE) on oxidized low-density lipoprotein (ox-LDL)-induced hyperpermeability in human umbilical vein endothelial cells(HUVECs).

Methods

HUVECs at passage 3 were divided into the control group and the ox-LDL(10

50 and 100 μg/mL) intervention groups (

=3) for the examination of transendothelial electrical resistance (TER). HUVECs cultured in slides were randomly divided into the control group

the ox-LDL 24 h group and the ox-LDL 48 h group. The distribution of F-actin and VE-cadherin were detected by immunofluorescence

and the protein content of VE-cadherin was detected by Western blotting. Incubated with calcium fluorescence probe (Fluo-3/AM)

intracellular free Ca

concentration (control group

ox-LDLgroup

ox-LDL+2-APB group) and SOCE (control group

ox-LDL 24 h group

ox-LDL 48 h group) were detected by laser confocal microscopy.

Results

Low dose of ox-LDL(10

25 μg/mL) exerted no effect on TER. High dose of ox-LDL (50

100 μg/mL) also had no effect on TER within 6 h

but after 6 h showed no effect on TER. However

high dose of ox-LDL(50

100 μg/mL) reduced the decrease of TER in 12 h (

0.05)

and the decrease was more significant in 24 h and 48 h. After the incubation with ox-LDL(100 μg/mL) 24 h and 48 h

the expression level of VE-cadherin decreased(

0.05)

while the form and distribution of F-action changed; Intracel

lular free Ca

concentration increased obviously which was interrupted by 2-APB the SOCE inhibitor (

0.05)

and the 2-APB can decrease the effect of ox-LDL on TER. After incubation with ox-LDL(100 μg/mL) 48 h

the amplitude of SOCE decreased (

0.05). The descending artery flow velocity of SOCE showed a slightly decreased trend after incubation with ox-LDL 24 h and a more significant decrease in 48 h(

0.05).

Conclusion

Ox-LDL up-regulate the intracellular free Ca

concentration of vascular endothelial cells by extending the duration of SOCE to effect the arrangement of cytoskeleton and cell junction

which increases the cell permeability.

关键词

Keywords

references

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