| 摘要: |
| 目的 探讨脂多糖诱导下血管内皮细胞热休克蛋白A12B(HSPA12B)表达的改变及其对血管内皮通透性的影响。 方法 体外传代培养人脐静脉内皮细胞, 并将其分为空白组(未进行任何处理)、脂多糖(1 μg/mL)诱导组、脂多糖+HSPA12B 高表达组(转染HSPA12B过表达质粒)和脂多糖+阴性质粒对照组(转染阴性对照质粒)。采用电阻抗检测仪检测脐静脉内皮细胞的跨内皮电阻抗, 流式细胞仪检测血管内皮钙黏素(VE-cadherin)的表达, 蛋白质印迹、PCR检测 HSPA12B 的表达改变。结果 脐静脉内皮细胞经脂多糖诱导后, 细胞中HSPA12B表达在0~12 h内上调, 12 h时达高峰。HSPA12B能使脐静脉内皮细胞的跨内皮电阻抗值明显升高(P<0.001), 并能完全对抗脂多糖诱导造成的跨内皮电阻抗值的下降(P<0.001)。HSPA12B能使脐静脉内皮细胞细胞膜上VE-cadherin的表达上调。结论 HSPA12B可能通过上调 VE-cadherin 表达降低血管内皮细胞的通透性, 从而保护血管内皮的屏障功能。 |
| 关键词: 热休克蛋白A12B 脂多糖类 人脐静脉内皮细胞 跨内皮电阻抗 |
| DOI：10.3724/SP.J.1008.2015.00483 |
| 投稿时间：2014-12-18修订日期：2015-03-02 |
| 基金项目:国家自然科学基金(81270128). |
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| Heat shock protein A12B decreases lipopolysaccharide-induced permeability of human umbilical vein endothelial cells |
| KANG Qiu-xiang1,CHEN Yi1,YU Gui-fang2,ZHANG Xu1*,ZHU Ke-ming1* |
(1. Department of Anesthesiology (Intensive Care Unit), Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Department of Anesthesiology, the 3rd People's Hospital of Shanghai, Shanghai Jiaotong University School of Medicine, Shanghai 201999, China
*Corresponding authors) |
| Abstract: |
| Objective To explore the lipopolysaccharide (LPS)-induced changes of heat shock protein A12B (HSPA12B) expression and its effect on the permeability of human umbilical vein endothelial cells (HUVECs). Methods The HUVECs cultured in vitro were divided into four groups: control group (without any treatment); LPS group (1 μg/mL LPS); LPS +pIRES2-EGFP-HSPA12B-3Flag group(The HSPA12B gene overexpression plasmid was transiently transfected into HUVECs and then LPS of 1 μg/mL was added); and LPS+pIRES2-EGFP-3Flag group (The negative control plasmid was transiently transfected into HUVECs and then LPS of 1 μg/mL was added). The transendothelial electrical resistance (TEER) of HUVECs was measured by MERSST×01 Electrode. The expression of VE-cadherin was studied by flow cytometry. RT-PCR and Western blotting analysis were used to examine the expression changes of HSPA12B mRNA and protein in HUVECs stimulated with LPS at various time points. Results It was showed that after stimulated with LPS, HSPA12B mRNA and protein were gradually upregulated, and peaked at 12 h. HSPA12B significantly increased the TEER value of HUVECs (P<0.001), and it could completely offset the decline of TEER value induced by LPS(P<0.001). Besides, HSPA12B could cause upregulation of VE-cadherin expression. Conclusion HSPA12B can reduce the permeability of vascular endothelial cells by up-regulating the expression of VE-cadherin, thus protecting the vascular endothelial barrier function of endothelial cells. |
| Key words: heat-shock protein A12B lipopolysaccharides human umbilical vein endothelial cells transendothelial electrical resistance |
