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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(1μg/m L)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。
Objective To investigate the changes of heat shock protein A12B (HSPA12B) expression in vascular endothelial cells induced by lipopolysaccharide and its effect on vascular endothelial permeability. Methods Human umbilical vein endothelial cells were subcultured in vitro and divided into blank group (without any treatment), lipopolysaccharide (1μg / ml) induction group, lipopolysaccharide + HSPA12B high expression group (HSPA12B transfection plasmid) And lipopolysaccharide + negative plasmid control group (transfected negative control plasmid). The trans-endothelial electrical impedance of umbilical vein endothelial cells was detected by electrical impedance spectroscopy. The expression of VE-cadherin was detected by flow cytometry. The expression of HSPA12B was detected by Western blot and PCR. Results After induced by lipopolysaccharide, the expression of HSPA12B in umbilical vein endothelial cells was upregulated within 0-12 h and peaked at 12 h. HSPA12B could significantly increase the trans-endothelial electrical impedance of human umbilical vein endothelial cells (P <0.001) and completely antagonize the decrease of trans-endothelial electrical impedance induced by lipopolysaccharide (P <0.001). HSPA12B can up-regulate the expression of VE-cadherin on the cell membrane of umbilical vein endothelial cells. Conclusion HSPA12B may reduce the permeability of vascular endothelial cells by up-regulating the expression of VE-cadherin, thereby protecting the barrier function of vascular endothelial cells.