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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。
Objective To observe the effect of high-pressure culture on endothelial cell E-selectin expression and its possible molecular mechanism. Methods Human umbilical vein endothelial cells (HUVECs) were cultured in a self-made adjustable pressure incubator at different pressures of 1 standard atmospheric pressure. The expression of E-selectin in vascular endothelial cells was detected by indirect immunofluorescence The expression of NF-κB inhibitor α was detected by Western blot. The expression of NF-κB was detected by indirect immunofluorescence staining. Results There was no significant difference in cell morphology between high-pressure culture and atmospheric pressure culture. The expression of E-selectin increased significantly in high-pressure culture, and the expression of E-selectin increased by 1.5-fold at 180 mmHg in 24 h. The expression of E-selectin at 180 mmHg for 6 h and 12 h The expression of NF-κB decreased by more than 2 folds. Activation of nuclear factor κB at 180 mmHg for 24 h reversed the nuclear translocation. Conclusion High-pressure culture can promote the expression of E-selectin and the regulation of NF-κB signaling pathway.