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目的探讨氧化低密度脂蛋白(ox-LDL)对人冠状动脉内皮细胞表达B7相关蛋白-1(B7RP-1)的影响及其临床意义。方法体外培养人冠状动脉内皮细胞,通过间接免疫荧光法、RT-PCR和Western blotting等方法,观察B7RP-1在人冠状动脉内皮细胞表达及ox-LDL的干预作用。结果(1)B7RP-1表达于人冠状动脉内皮细胞,荧光信号呈点状,散在分布于细胞表面;RT-PCR显示B7RP-1的mRNA扩增产物片段位于相当于Marker 469bp的位置,Western blotting检测B7RP-1的相对分子质量大小约为70000。(2)ox-LDL刺激组B7RP-1的平均光密度OD值和吸光度A值均分别高于空白对照组,ox-LDL可增加B7RP-1在人冠状动脉内皮细胞中的mRNA和蛋白表达,并具有统计学意义(均P<0.05)。结论B7RP-1表达于人冠状动脉内皮细胞表面,具有慢性致炎症作用的ox-LDL可上调共刺激分子B7RP-1 mRNA和蛋白表达,这可能是动脉粥样硬化的免疫学发病机制之一。
Objective To investigate the effect of ox-LDL on the expression of B7-related protein-1 (B7RP-1) in human coronary artery endothelial cells and its clinical significance. Methods Human coronary artery endothelial cells were cultured in vitro. The expression of B7RP-1 in human coronary artery endothelial cells and the intervention of ox-LDL were detected by indirect immunofluorescence, RT-PCR and Western blotting. Results (1) B7RP-1 was expressed in human coronary artery endothelial cells, the fluorescence signal was dotted and scattered on the cell surface; RT-PCR showed that the fragment of mRNA amplification product of B7RP-1 was located at the position corresponding to 469bp of Marker, The relative molecular mass of B7RP-1 was about 70,000. (2) The OD value and absorbance A value of B7RP-1 in ox-LDL stimulation group were higher than that in blank control group, ox-LDL increased the mRNA and protein expression of B7RP-1 in human coronary artery endothelial cells, And had statistical significance (all P <0.05). Conclusions B7RP-1 is expressed on the surface of human coronary artery endothelial cells. Ox-LDL with chronic inflammation may up-regulate costimulatory molecules B7RP-1 mRNA and protein expression, which may be one of the immunological pathogenesis of atherosclerosis.