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目的:研究人类单核细胞系U937经氧化低密度脂蛋白(ox-LDL)诱导成泡沫细胞后,血管内皮生长因子(VEGF)的表达水平,观测反义寡脱氧核苷酸(asODN)对VEGF表达的抑制作用。方法:U937细胞与80mg/L ox-LDL孵育48h后,加入asODN(0,5,10,20μmol/L)。采用基础酶联免疫吸附分析法(ELISA)检测各组培养基中VEGF蛋白分泌含量。用免疫组织化学检测VEGF蛋白表达,采用CA6300型图像分析系统分析切片阳性面积比来衡量蛋白表达水平;用RNA印迹法检测VEGF mRNA水平。结果:培养的U937泡沫细胞中VEGF表达均明显高于U937正常细胞。asODN可显著降低泡沫细胞中VEGF的表达。经asODN 20μmol/L作用后,细胞培养液中VEGF含量降低45.0％,免疫组化显示细胞中VEGF阳性率降低64.9％,VEGF mRNA水平降低了47.1％。结论:泡沫细胞中VEGF表达明显增加,asODN能显著抑制U937泡沫细胞中VEGF的表达。
OBJECTIVE: To study the expression of vascular endothelial growth factor (VEGF) in human monocytic cell line U937 induced by ox-LDL and to observe the effect of asODN on VEGF Inhibition of expression. Methods: After U937 cells incubated with 80 mg / L ox-LDL for 48 h, asODN (0, 5, 10 and 20 μmol / L) was added. The basal enzyme-linked immunosorbent assay (ELISA) was used to detect the secretion of VEGF protein in each group. The expression of VEGF protein was detected by immunohistochemistry. The expression of VEGF protein was detected by CA6300 image analysis system. The VEGF mRNA level was detected by Northern blotting. Results: The expression of VEGF in cultured U937 foam cells was significantly higher than that in U937 normal cells. asODN can significantly reduce the expression of VEGF in foam cells. After treated with asODN 20μmol / L, the VEGF content in the cell culture medium decreased by 45.0%. Immunohistochemistry showed that the VEGF positive rate in the cells decreased by 64.9% and the level of VEGF mRNA decreased by 47.1%. Conclusion: The expression of VEGF in foam cells increased significantly, asODN could significantly inhibit the expression of VEGF in U937 foam cells.