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Store-operated Ca2+ entry plays a role in HMGB1-induced vascular endothelial cell hyperpermeability.

Zou M, Dong H, Meng X, Cai C, Li C, Cai S, Xue Y - PLoS ONE (2015)

Bottom Line: We have shown that human vascular endothelial cell permeability is increased, while transendothelial electrical resistance and VE-cadherin expression were reduced by HMGB1 treatment.Two SOCE inhibitors and knockdown of stromal interaction molecule 1 (STIM1), a Ca2+ sensor mediating SOCE, inhibited the HMGB1-induced influx of Ca2+ and Src activation followed by significant suppression of endothelial permeability.Moreover, knockdown of Orai1, an essential pore-subunit of SOCE channels, decreased HMGB1-induced endothelial hyperpermeability.

View Article: PubMed Central - PubMed

Affiliation: Department of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China.

ABSTRACT

Aims: Endothelial dysfunction, including increased endothelial permeability, is considered an early marker for atherosclerosis. High-mobility group box 1 protein (HMGB1) and extracellular Ca2+ entry, primarily mediated through store-operated Ca2+ entry (SOCE), are known to be involved in increasing endothelial permeability. The aim of this study was to clarify how HMGB1 could lead to endothelia hyperpermeability.

Methods and results: We have shown that human vascular endothelial cell permeability is increased, while transendothelial electrical resistance and VE-cadherin expression were reduced by HMGB1 treatment. Two SOCE inhibitors and knockdown of stromal interaction molecule 1 (STIM1), a Ca2+ sensor mediating SOCE, inhibited the HMGB1-induced influx of Ca2+ and Src activation followed by significant suppression of endothelial permeability. Moreover, knockdown of Orai1, an essential pore-subunit of SOCE channels, decreased HMGB1-induced endothelial hyperpermeability.

Conclusions: These data suggest that SOCE, acting via STIM1, might be the predominant mechanism of Ca2+ entry in the modulation of endothelial cell permeability. STIM1 may thus represent a possible new therapeutic target against atherosclerosis.

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Related in: MedlinePlus

HMGB1 increases endothelial cell permeability.A. HMGB1-induced TEER decrease. EA.hy926 cells cultured on transwell filters were incubated for 3, 6, 12, 24, 36 and 48h, respectively, with or without 50, 100, 200 and 400 ng/ml HMGB1. The integrity of the tight junctions was assessed by measuring the TER. B. Cell viability in the cells treated by HMGB1. EA.hy926 cells were treated with 50, 100, 200, 400 and 800ng/ml HMGB1, respectively, for 24 h. The cell viability was measured by CCK-8 assay. Data are presented as mean ± SD of three independent experiments. *Indicates significant difference compared with the control group (P<0.05).
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pone.0123432.g001: HMGB1 increases endothelial cell permeability.A. HMGB1-induced TEER decrease. EA.hy926 cells cultured on transwell filters were incubated for 3, 6, 12, 24, 36 and 48h, respectively, with or without 50, 100, 200 and 400 ng/ml HMGB1. The integrity of the tight junctions was assessed by measuring the TER. B. Cell viability in the cells treated by HMGB1. EA.hy926 cells were treated with 50, 100, 200, 400 and 800ng/ml HMGB1, respectively, for 24 h. The cell viability was measured by CCK-8 assay. Data are presented as mean ± SD of three independent experiments. *Indicates significant difference compared with the control group (P<0.05).

Mentions: HMGB1 has been implicated in the progression of atherosclerosis [29]. HMGB1-treated vascular endothelial cells were used to investigate the mechanisms underlying atherosclerosis. We measured transendothelial electric resistance (TER) to clarify the influence of HMGB1 on permeability. 50 and 100ng/ml of HMGB1 has no effect, however, 200 ng/ml of HMGB1 caused a decrease in TER at 6 h of treatment and a significant drop at 12–48 h of treatment, in a time-dependent manner. 400 ng/ml of HMGB1 decreased TER starting from 6h (Fig 1A). Exposing the cells to 50–800 ng/ml of HMGB1 for 24h had no impact on cell viability, implying that the increase in permeability was not due to inhibition of cell viability (Fig 1B).


Store-operated Ca2+ entry plays a role in HMGB1-induced vascular endothelial cell hyperpermeability.

Zou M, Dong H, Meng X, Cai C, Li C, Cai S, Xue Y - PLoS ONE (2015)

HMGB1 increases endothelial cell permeability.A. HMGB1-induced TEER decrease. EA.hy926 cells cultured on transwell filters were incubated for 3, 6, 12, 24, 36 and 48h, respectively, with or without 50, 100, 200 and 400 ng/ml HMGB1. The integrity of the tight junctions was assessed by measuring the TER. B. Cell viability in the cells treated by HMGB1. EA.hy926 cells were treated with 50, 100, 200, 400 and 800ng/ml HMGB1, respectively, for 24 h. The cell viability was measured by CCK-8 assay. Data are presented as mean ± SD of three independent experiments. *Indicates significant difference compared with the control group (P<0.05).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4401536&req=5

pone.0123432.g001: HMGB1 increases endothelial cell permeability.A. HMGB1-induced TEER decrease. EA.hy926 cells cultured on transwell filters were incubated for 3, 6, 12, 24, 36 and 48h, respectively, with or without 50, 100, 200 and 400 ng/ml HMGB1. The integrity of the tight junctions was assessed by measuring the TER. B. Cell viability in the cells treated by HMGB1. EA.hy926 cells were treated with 50, 100, 200, 400 and 800ng/ml HMGB1, respectively, for 24 h. The cell viability was measured by CCK-8 assay. Data are presented as mean ± SD of three independent experiments. *Indicates significant difference compared with the control group (P<0.05).
Mentions: HMGB1 has been implicated in the progression of atherosclerosis [29]. HMGB1-treated vascular endothelial cells were used to investigate the mechanisms underlying atherosclerosis. We measured transendothelial electric resistance (TER) to clarify the influence of HMGB1 on permeability. 50 and 100ng/ml of HMGB1 has no effect, however, 200 ng/ml of HMGB1 caused a decrease in TER at 6 h of treatment and a significant drop at 12–48 h of treatment, in a time-dependent manner. 400 ng/ml of HMGB1 decreased TER starting from 6h (Fig 1A). Exposing the cells to 50–800 ng/ml of HMGB1 for 24h had no impact on cell viability, implying that the increase in permeability was not due to inhibition of cell viability (Fig 1B).

Bottom Line: We have shown that human vascular endothelial cell permeability is increased, while transendothelial electrical resistance and VE-cadherin expression were reduced by HMGB1 treatment.Two SOCE inhibitors and knockdown of stromal interaction molecule 1 (STIM1), a Ca2+ sensor mediating SOCE, inhibited the HMGB1-induced influx of Ca2+ and Src activation followed by significant suppression of endothelial permeability.Moreover, knockdown of Orai1, an essential pore-subunit of SOCE channels, decreased HMGB1-induced endothelial hyperpermeability.

View Article: PubMed Central - PubMed

Affiliation: Department of Endocrinology and Metabolism, Nanfang Hospital, Southern Medical University, Guangzhou, 510515, China.

ABSTRACT

Aims: Endothelial dysfunction, including increased endothelial permeability, is considered an early marker for atherosclerosis. High-mobility group box 1 protein (HMGB1) and extracellular Ca2+ entry, primarily mediated through store-operated Ca2+ entry (SOCE), are known to be involved in increasing endothelial permeability. The aim of this study was to clarify how HMGB1 could lead to endothelia hyperpermeability.

Methods and results: We have shown that human vascular endothelial cell permeability is increased, while transendothelial electrical resistance and VE-cadherin expression were reduced by HMGB1 treatment. Two SOCE inhibitors and knockdown of stromal interaction molecule 1 (STIM1), a Ca2+ sensor mediating SOCE, inhibited the HMGB1-induced influx of Ca2+ and Src activation followed by significant suppression of endothelial permeability. Moreover, knockdown of Orai1, an essential pore-subunit of SOCE channels, decreased HMGB1-induced endothelial hyperpermeability.

Conclusions: These data suggest that SOCE, acting via STIM1, might be the predominant mechanism of Ca2+ entry in the modulation of endothelial cell permeability. STIM1 may thus represent a possible new therapeutic target against atherosclerosis.

Show MeSH
Related in: MedlinePlus