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目的:研究溶血磷脂酰胆碱(LPC)对大鼠主动脉内皮细胞(RAEC)中血管内皮生长因子(VEGF)表达的影响以及银杏叶提取物(Ginkgo bilobaextract,GbE)对RAEC的保护作用.方法:MTT法和LDH的释放检测RAEC的损伤;用基础酶联免疫吸附实验(ELISA)检测VEGF蛋白含量;用RNA点杂交(Dot blot)法和原位杂交法检测细胞中 VEGF mRNA的表达.结果:LPC 5 mg/L明显抑制RAEC的生长,加入 GbE 0.01-1 μg/L后,LPC对RAEC生长抑制率明显降低。LPC可使RAEC条件培养基中VEGF蛋白表达明显增加,GbE可剂量依赖性地降低VEGF的蛋白含量.原位杂交及点杂交结果显示正常培养的RAEC未见明显的 VEGF mRNA表达,LPC刺激后可见其高表达,同时加入GbE后,VEGF mRNA的表达明显降低.结论:LPC能诱导RAEC表达高水平的VEGF,GbE可明显降低其含量.
Objective: To study the effect of lysophosphatidylcholine (LPC) on the expression of vascular endothelial growth factor (VEGF) in rat aortic endothelial cells (RAEC) and the protective effect of Ginkgo biloba extract (GbE) on RAEC. : MTT assay and LDH release detect RAEC damage; use basic enzyme-linked immunosorbent assay (ELISA) to detect VEGF protein content; use RNA dot blot (Dot blot) and in situ hybridization to detect VEGF mRNA expression in cells. :LPC 5 mg/L significantly inhibited the growth of RAEC. After addition of GbE 0.01-1 μg/L, LPC inhibited the growth of RAEC significantly. LPC can increase the expression of VEGF protein in RAEC conditioned media, and GbE can decrease the protein content of VEGF in a dose-dependent manner. The results of in situ hybridization and dot blot showed that there was no obvious VEGF mRNA expression in normal cultured RAEC, which could be seen after LPC stimulation. The expression of VEGF mRNA was significantly decreased after adding GbE. Conclusion: LPC can induce high expression of VEGF in RAEC, and GbE can significantly reduce its content.