高静水压培养对内皮酯酶表达的影响

Effect of High Hydrostatic Pressure on Endothelial Lipase Expression

探讨高静水压培养对内皮脂酶表达的影响及其机制.分大气压(0mmHg)和高于1个标准大气压的递增压力(120、150、180mmHg)培养人脐静脉内皮细胞,用蛋白酶体抑制剂MG132进行干预.RT-PCR检测内皮脂酶mRNA的表达,间接免疫荧光技术和流式细胞术检测内皮脂酶蛋白的表达.结果显示,高静水压培养明显促进内皮脂酶mRNA和蛋白质的表达,180mmHg培养24h内皮脂酶mRNA的表达较对照组上调2.2倍(P<0.001),内皮脂酶蛋白表达较对照组上调2.54倍(P<0.001).蛋白酶体抑制剂MG132干预明显抑制180mmHg培养诱导的内皮脂酶mRNA的表达,MG132干预组内皮脂酶mRNA表达约为180mmHg培养组的50%(P<0.05).结果说明,高静水压上调内皮脂酶mRNA和蛋白质的表达,其机制可能与核因子-κB活化有关.

Hydrostatic pressure has direct effect on vascular endothelium. Endothelial lipase （EL） is a newly identified member, only secreted by endothelial cells, of the triglycerides lipase family. EL may be one important determinants of HDL-metabolism and inflammation acting at the vessel wall. However, little is known about regulation of EL at the vessel wall. The effect of hydrostatic pressure on EL and its possible mechanism were investigated. In human umbilical vein endothelial cells, increasing pressures （120, 150 and 180 mmHg） treatment in a custom-made pressure incubator increased the mRNA and protein expression level of EL without obvious cells apoptosis.180 mmHg treatment up-regulated EL mRNA and protein about 2.2, 2.54 fold relative to pressure atmosphere, respectively. Inhibitor of nuclear factor-κB MG132 attenuated the high pressure induction of EL expression level, which is half of EL expression in 180 mmHg treatment group. These findings provide new insights into the effect of hydrostatic pressure on EL expression through nuclear factor-κB signal pathways.