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目的研究熊果酸(Ursolic acid,UA)对人脐静脉血管内皮细胞(Human umbilical vein endothelial cells,HUVECs)增殖及ERK1、c-Jun、c-Myc、Cyclin D1基因mRNA和蛋白表达的影响,探讨熊果酸抑制血管生长的机制。方法体外培养HU-VECs,用不同浓度熊果酸(31.5、62.5、125、250、500μg/ml)处理不同时间(12、24、48 h),MTT法检测细胞增殖抑制率。用125μg/ml熊果酸和100μmol/L PD98059(ERK抑制剂)处理HUVECs 48 h;不同浓度熊果酸处理24 h;125μg/ml熊果酸处理不同时间,RT-PCR和Western blot分别检测HUVECs中ERK1、c-Jun、c-Myc、Cyclin D1基因mRNA的转录水平和蛋白的表达水平。结果不同浓度的熊果酸对HUVECs的增殖均有明显的抑制作用,且呈剂量和时间依赖性;经熊果酸和PD98059处理的HUVECs中ERK1、c-Jun、c-Myc、Cyclin D1基因mRNA和蛋白的表达水平均明显降低,且呈剂量和时间依赖性。结论熊果酸可通过抑制ERK信号转导通路而抑制血管内皮细胞增殖。
Objective To investigate the effects of Ursolic acid (UA) on the proliferation of human umbilical vein endothelial cells (HUVECs) and the mRNA and protein expressions of ERK1, c-Jun, c-Myc and Cyclin D1 Mechanism of ursolic acid inhibiting blood vessel growth. Methods HU-VECs were cultured in vitro. UA (31.5, 62.5, 125, 250 and 500μg / ml) was used to treat HU-VECs at different times (12,24,48 h) HUVECs were treated with 125μg / ml ursolic acid and 100μmol / L PD98059 (ERK inhibitor) for 48 h. UA at different concentrations for 24 h and 125 μg / ml UA at different times. HUVECs were detected by RT-PCR and Western blot respectively The mRNA and protein expression levels of ERK1, c-Jun, c-Myc, and Cyclin D1 were detected by Western blot. Results UA at different concentrations significantly inhibited the proliferation of HUVECs in a dose and time dependent manner. The expressions of ERK1, c-Jun, c-Myc and Cyclin D1 mRNA in UUVECs treated with ursolic acid and PD98059 And protein expression levels were significantly reduced, and in a dose and time-dependent manner. Conclusion Ursolic acid can inhibit the proliferation of vascular endothelial cells by inhibiting ERK signal transduction pathway.