内皮细胞表达血管内皮细胞生长因子的研究

Expression of vascular endothelial growth factor in endothelial cells: an in vitro study

目的 观察体外培养的内皮细胞表达和分泌血管内皮细胞生长因子 (VEGF)的情况。方法 以脐静脉内皮细胞系ECV 30 4为研究对象 ,采用RT PCR、流式细胞仪、ELISA法分别观察佛波脂 (PMA)对ECV 30 4合成与分泌VEGF的作用。结果 10 0ng/mlPMA能明显上调VEGFmRNA的表达、VEGF蛋白的合成及分泌。其对VEGFmRNA表达及细胞内VEGF合成的作用在 12h最为明显 ;而分泌至上清液中的VEGF量则在 18h达到高峰。PMA的上述作用能够被放线菌素D所抑制。结论 PMA能够诱导内皮细胞ECV 30 4表达、合成及分泌VEGF ,该作用呈剂量和时间依赖性。其对VEGFmRNA的作用涉及核苷酸的从头合成途径。

Objective To observe the expression and production of vascular endothelial growth factor (VEGF) induced by PMA in a human umbilical vein endothelial derived cell line ECV-304 according to American Tissue and Cell Collection(ATCC) so as to set up a convenient and stable model for the study of endothelial function. Methods RT-PCR was used to detect VEGF mRNA expression, while flow cytometry combined with specific anti-VEGF antibody to measure intracellular VEGF protein production. ELISA was utilized to determine the VEGF concentration secreted by ECV-304. Results It was found that 100 ng/ml PMA, the optimal concentration determined in the present study, increased markedly the VEGF mRNA expression after 2 h incubation. This effect lasted for at least 12 hours and reached the maximum by 12 h Actinomycin D depressed the effect of PMA. The ELISA data demonstrated that the secreted VEGF protein level reached the maximum by 18 h. Conclusions PMA can induce VEGF expression and production in ECV-304 deposeding on dose-and time involving the whole synthetic processing of the VEGF mRNA.