普通肝素改善高迁移率族蛋白1介导的血管内皮细胞屏障通透性增加的实验研究
发布时间：2018-10-14 18:37
【摘要】：目的 观察普通肝素(UFH)对高迁移率族蛋白1(HMGB 1)介导的人脐静脉内皮细胞屏障通透性损伤的保护作用,探讨UFH对HMGB 1介导的胞质紧密粘连蛋白-1(ZO-1)表达缺失的保护机制。方法 将人脐静脉内皮细胞进行体外培养,人脐静脉内皮细胞株传代培养后分为4组(n=5):空白对照组(加入等量PBS)、HMGB 1处理组(100 ng/ml)、UFH对照组(UFH 10 U/ml)、HMGB 1及UFH处理组(100 ng/ml HMGB 1+UFH 10 U/ml)。MTT法测定内皮细胞存活率,用Transwell小室法测定单层内皮屏障通透性,用免疫荧光染色法测定ZO-1的表达分布,蛋白免疫印迹(Western blot)法检测ZO-1蛋白及核因子-κB(NF-κB)的表达。结果 HMGB 1(100 ng/ml)对内皮细胞活性无抑制作用(P0.05)。UFH预处理后可减少HMGB1所致的内皮细胞通透性增加(P0.05)。UFH预处理后可降低HMGB 1所致内皮细胞ZO-1密闭环的减少和破坏,使ZO-1荧光强度增强,ZO-1蛋白表达增加,并使NF-κB的核易位减少。结论 UFH可保护HMGB 1介导的内皮细胞ZO-1的表达缺失,进而改善内皮细胞屏障通透性,其机制与减少NF-κB核易位相关。
[Abstract]:Objective to investigate the protective effect of common heparin (UFH) on barrier permeability injury of human umbilical vein endothelial cells mediated by high mobility group protein 1 (HMGB 1), and to explore the protective mechanism of UFH against HMGB 1 mediated loss of cytoplasmic compact adhesion protein-1 (ZO-1) expression. Methods Human umbilical vein endothelial cells were cultured in vitro. Human umbilical vein endothelial cells (HUVEC) were divided into 4 groups after subculture: blank control group (UFH 10 U/ml), HMGB 1) and UFH treated group (100 ng/ml HMGB 1 UFH 10 U/ml). MTT). The permeability of endothelial barrier was measured by Transwell chamber method, the expression of ZO-1 was detected by immunofluorescence staining, and the expression of ZO-1 protein and nuclear factor- 魏 B (NF- 魏 B) were detected by Western blot (Western blot). Results HMGB 1 (100 ng/ml) had no inhibitory effect on endothelial cell activity (P0.05). UFH pretreatment decreased the increase of endothelial cell permeability induced by HMGB1 (P0.05). UFH pretreatment decreased the decrease and destruction of ZO-1 closed loop induced by HMGB 1. The fluorescence intensity of ZO-1 was increased, the expression of ZO-1 protein was increased, and the nuclear translocation of NF- 魏 B was decreased. Conclusion UFH can protect HMGB 1 mediated ZO-1 expression deletion in endothelial cells and improve the permeability of endothelial cell barrier. The mechanism is related to the reduction of NF- 魏 B translocation.
【作者单位】： 中国医科大学附属第一医院重症医学科;
[Abstract]:Objective to investigate the protective effect of common heparin (UFH) on barrier permeability injury of human umbilical vein endothelial cells mediated by high mobility group protein 1 (HMGB 1), and to explore the protective mechanism of UFH against HMGB 1 mediated loss of cytoplasmic compact adhesion protein-1 (ZO-1) expression. Methods Human umbilical vein endothelial cells were cultured in vitro. Human umbilical vein endothelial cells (HUVEC) were divided into 4 groups after subculture: blank control group (UFH 10 U/ml), HMGB 1) and UFH treated group (100 ng/ml HMGB 1 UFH 10 U/ml). MTT). The permeability of endothelial barrier was measured by Transwell chamber method, the expression of ZO-1 was detected by immunofluorescence staining, and the expression of ZO-1 protein and nuclear factor- 魏 B (NF- 魏 B) were detected by Western blot (Western blot). Results HMGB 1 (100 ng/ml) had no inhibitory effect on endothelial cell activity (P0.05). UFH pretreatment decreased the increase of endothelial cell permeability induced by HMGB1 (P0.05). UFH pretreatment decreased the decrease and destruction of ZO-1 closed loop induced by HMGB 1. The fluorescence intensity of ZO-1 was increased, the expression of ZO-1 protein was increased, and the nuclear translocation of NF- 魏 B was decreased. Conclusion UFH can protect HMGB 1 mediated ZO-1 expression deletion in endothelial cells and improve the permeability of endothelial cell barrier. The mechanism is related to the reduction of NF- 魏 B translocation.
【作者单位】： 中国医科大学附属第一医院重症医学科;
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