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目的:研究溶血磷脂酰胆碱(LPC)对内皮细胞(EC)中血管内皮生长因子(VEGF)表达的影响。方法:在人脐静脉内皮细胞株ECV04培养基中加入不同浓度的LPC,培养不同时间,用基础酶联免疫吸附实验(ELISA)检测各组EC条件培养基中VEGF蛋白含量;用免疫组织化学法检测EC中VEGF蛋白及其受体的表达;用原位杂交检测VEGF信使核糖核酸(VEGF mRNA)的表达。结果:培养的ECV304能表达VEGF受体,在胞浆内呈棕色颗粒,当LPC刺激后,阳性增强。原位杂交结果显示,培养的ECV304中未见VEGF mRNA的表达,当LPC刺激后可见VEGF mRNA的高表达,在胞浆内呈棕色颗粒。ELISA结果显示LPC可使ECV304条件培养基中VEGF蛋白含量明显增加,且具有时间和剂量依赖性。结论:LPC能诱导ECV304表达高水平的VEGF。 (责任编辑 吕静)
Objective: To investigate the effect of lysophosphatidylcholine (LPC) on the expression of vascular endothelial growth factor (VEGF) in endothelial cells (ECs). Methods: Different concentrations of LPC were added to ECV04 medium of human umbilical vein endothelial cells and cultured for different time. The content of VEGF protein in EC conditioned medium was detected by enzyme-linked immunosorbent assay (ELISA). Immunohistochemistry The expression of VEGF protein and its receptor was detected in EC. The expression of VEGF mRNA was detected by in situ hybridization. Results: The cultured ECV304 could express VEGF receptor and showed brown granules in the cytoplasm. When stimulated by LPC, it was positive. In situ hybridization results showed that the expression of VEGF mRNA was not observed in cultured ECV304 cells. When stimulated with LPC, the expression of VEGF mRNA was high and brown granules were found in the cytoplasm. ELISA results showed that LPC can significantly increase the VEGF protein content of ECV304 conditioned medium in a time-and dose-dependent manner. Conclusion: LPC can induce ECV304 to express high level of VEGF. (Editor LV Jing)