高压培养调节E选择素表达及核因子κB信号通路

High Pressure Regulating the Expressions of E-Selectin and Nuclear Factor KappaB Signaling Pathway

目的观察高压培养对内皮细胞E选择素表达的影响及其可能的分子机制。方法在自制的可调压力培养箱中,用高于1个标准大气压的不同压力培养人脐静脉内皮细胞,运用间接免疫荧光技术、流式细胞术检测血管内皮细胞粘附分子E选择素蛋白的表达,Western blot检测核因子κB抑制因子α的表达,间接免疫荧光法观察核因子κB的表达分布。结果高压培养和大气压培养细胞形态无明显差异;高压培养E选择素表达明显增加,180 mmHg培养24 h E选择素的表达升高1.5倍;180 mmHg培养6 h和12 h核因子κB抑制因子α的表达降低2倍多;180 mmHg培养24 h激活核因子κB,使其转位于细胞核。结论高压培养促进E选择素表达,核因子κB信号通路参与调控。

Aim To investigate the effect and possible mechanisms of high pressure on the expression of E-selectin in vascular endothelial cells. Methods Human umbilical vein endothelial cells were cultured under 0, 120, 150, 180, 210 mmHg in a custom-made pressure incubator for 24 h or treated under 180 mmHg of static pressure for 0, 3, 6, 12, 24h. Western blot, flow cytomety and immunofluorescence were used to detect the expression of E-selectin, inhibitor of nuclear factor kappaB alpha （ IκBa） and nuclear factor kappaB （ NF-κB ） respectively. Results High pressure significantly up-regulated the level of E-selectin. E-selectin level was increased about 1.5 times at 180 mmHg for 24 h, IκBa was downregulated more than 2 times following 180 mmHg culturing for 6 h and 12 h, NF-κB was transferred from cytoplasm to nuclear by 180 mmHg culture for 24 h. Condusion High pressure significantly upregulates E-selectin expression by activating the NF-κB signaling pathway.