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目的探讨乌司他丁对脂多糖(LPS)诱发的单层人脐静脉内皮细胞(HUVECs)高通透性的影响及其可能机制。方法体外培养的单层HUVECs被分为对照组、LPS组(LPS 1μg/ml刺激4h)、乌司他丁复合LPS组(乌司他丁3000U/ml孵育1h+LPS 1μg/ml刺激4h)和血管内皮钙黏蛋白单克隆抗体(VE-cadherin mAb)组(VE-cadherin mAb 50μg/ml孵育6h+乌司他丁3000U/ml孵育1h+LPS 1μg/ml刺激4h)。采用双腔系统Transwell法检测单层HUVECs的通透性,Western blot法检测HUVECs中VE-cadherin的表达水平。结果与对照组相比,LPS组单层HUVECs渗透性增高,VE-cadherin表达降低(P<0.01)。与LPS组相比,乌司他丁复合LPS组单层HUVECs渗透性降低,VE-cadherin表达增高(P<0.01)。与乌司他丁复合LPS组相比,VE-cadherin mAb组单层HUVECs渗透性增高,VE-cadherin表达降低(P<0.01)。结论乌司他丁预处理可改善LPS引起的单层HUVECs高通透性状况;其可能机制与VE-cadherin表达增多有关。
Objective To investigate the effect of ulinastatin on the high permeability of monolayer human umbilical vein endothelial cells (HUVECs) induced by lipopolysaccharide (LPS) and its possible mechanism. Methods Monolayers of HUVECs cultured in vitro were divided into control group, LPS group (stimulated by LPS 1μg / ml for 4 hours), ulinastatin combined LPS group (stimulated by ulinastatin 3000U / ml for 1 hour + LPS 1μg / ml for 4 hours) and VE-cadherin mAb group was incubated with 50μg / ml VE-cadherin mAb for 6h + ulinastatin 3000U / ml for 1h + LPS 1μg / ml for 4h. The permeability of monolayer HUVECs was detected by dual-chamber system Transwell method. The expression of VE-cadherin in HUVECs was detected by Western blot. Results Compared with the control group, the permeability of monolayer HUVECs in LPS group was increased and the expression of VE-cadherin was decreased (P <0.01). Compared with LPS group, the permeability of monolayer HUVECs in ulinastatin LPS group decreased and the expression of VE-cadherin increased (P <0.01). Compared with ulinastatin complex LPS group, monolayer HUVECs in VE-cadherin mAb group had higher permeability and lower expression of VE-cadherin (P <0.01). Conclusion Ulinastatin pretreatment can improve the high permeability of monolayer HUVECs induced by LPS. The possible mechanism is related to the increased expression of VE-cadherin.