Gap Junction Channel Modulates Pulmonary Vascular Permeability through Calcium in Acute Lung Injury: An Experimental Study
Gap Junction Channel Modulates Pulmonary Vascular Permeability through Calcium in Acute Lung Injury: An Experimental Study
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DOI：
10.1159/000274384
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发表时间：
2010-01-01
期刊：
影响因子：
3.7
通讯作者：
Yi, Dinghua
中科院分区：
文献类型：
--
作者：
Zhang, Jinzhou;Wang, Wen;Yi, Dinghua
背景：肺血管通透性增加是急性肺损伤（ALI）的标志。间隙连接通道 (GJC) 连接相邻细胞并促进离子交换。目前尚不清楚 GJC 是否通过细胞内钙调节 ALI 的肺通透性。目的：本研究旨在验证肺微血管内皮细胞 (PMVEC) 中的 GJC 是否通过细胞内钙调节 ALI 中的肺血管通透性。方法：首先，使用连接蛋白 40 (Cx40) 免疫组织化学方法研究肺中埃文斯蓝 (EB) 渗漏的 ALI 动物模型。然后将培养的PMVEC分为三组：G（对照）、G（血清）和G（阻断剂）。血清取自枪伤后患有 ALI 的动物（受伤血清）。最初，用GJCs阻断剂处理G（阻断剂），然后分别用受损血清刺激G（血清）和G（阻断剂）。评估 GJC、细胞单层的渗透性和细胞内 Ca2+。结果：Cx40 随时间下降，而 EB 渗漏增加。 Cx40 和 EB 泄漏表现出很强的负相关性 (rho = -0.934, p < 0.05)。受伤的血清减少了 GJC 和 Cx40 的表达，而阻断剂则加剧了这种效应。同样，当用受损血清处理 PMVEC 单层时，通透性和细胞内 Ca2+ 均增加。阻滞剂也加剧了这些影响。结论：ALI 时 PMVEC 的 GJC 抑制增加了肺血管通透性；这种效应可能是由细胞内钙超载介导的。版权所有 (C) 2010 S. Karger AG，巴塞尔
Background: Increased pulmonary vascular permeability is a hallmark of acute lung injury (ALI). Gap junction channels (GJCs) connect adjacent cells and facilitate ion exchange. It remained unclear whether GJCs modulate pulmonary permeability in ALI through intracellular calcium. Objectives: This study aimed to verify if GJCs in pulmonary microvessel endothelial cells (PMVECs) modulate pulmonary vascular permeability in ALI via intracellular calcium. Methods: Firstly, an animal model of ALI was studied using connexin 40 (Cx40) inmunohistochemistry in the lung with Evans' blue (EB) leakage. Then cultured PMVECs were divided into three groups: G(control), G(serum) and G(blocker). Serum was obtained from animals with ALI following gunshot injury (injured serum). Initially, G(blocker) was treated with the blocker of GJCs, and then G(serum) and G(blocker) were stimulated with the injured serum, respectively. GJCs, the permeability of cell monolayers and intracellular Ca2+ were assessed. Results: Cx40 time-dependently decreased, whereas EB leakage increased. Cx40 and EB leakage exhibited a strong inverse correlation (rho = -0.934, p < 0.05). Injured serum decreased GJCs and expression of Cx40, whereas the blocker aggravated this effect. Similarly, when PMVEC monolayer was treated with injured serum, both permeability and intracellular Ca2+ increased. These effects were also aggravated with the blocker. Conclusions: Depression of GJCs of PMVECs increased pulmonary vascular permeability in ALI; this effect may be mediated by the overload of intracellular calcium. Copyright (C) 2010 S. Karger AG, Basel