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目的观察藏红花素(crocin)对人脐静脉内皮细胞(HUVECs)体外增殖、细胞外调节蛋白激酶(ERK)1/2表达及活性的影响。方法用藏红花素及MAPK/ERK激酶(MEK)抑制剂PD98059分别处理HUVECs,Ed U细胞增殖法检测细胞体外增殖活力,Western blotting法检测crocin对磷酸化细胞外调节蛋白激酶1/2(ERK1/2)和总ERK1/2表达的影响,激光扫描共焦显微镜检测细胞内Ca~(2+)浓度。结果在1μmol/L和10μmol/L浓度水平,藏红花素可明显促进细胞的增殖能力,提高磷酸化ERK1/2和总ERK1/2蛋白的表达水平,并增加细胞内Ca~(2+)浓度;给予MEK抑制剂PD98059后,细胞的增殖能力下降,磷酸化ERK1/2和总ERK1/2的表达减弱,细胞内Ca~(2+)浓度降低。结论藏红花素通过活化ERK1/2信号途径,提高细胞内Ca~(2+)浓度,促进人脐静脉内皮细胞的体外增殖能力。
Objective To observe the effects of crocin on the proliferation and the expression of extracellular regulated protein kinase (ERK) 1/2 in human umbilical vein endothelial cells (HUVECs). Methods HUVECs were treated with crocetin and MAPK / ERK kinase (MEK) inhibitor PD98059 respectively. Proliferation assay was used to detect the proliferative activity of cells in vitro by Ed U cell proliferation assay. The effect of crocin on phosphorylated extracellular regulated protein kinase 1/2 (ERK1 / 2) ) And total ERK1 / 2 expression. Laser scanning confocal microscopy was used to detect intracellular Ca2 + concentration. Results Crocetin obviously enhanced the proliferation of cells, increased the expression of phosphorylated ERK1 / 2 and total ERK1 / 2 protein and increased the intracellular Ca2 + concentration at the concentration of 1μmol / L and 10μmol / L. After PD98059, a MEK inhibitor, the proliferation of cells decreased, the expression of phosphorylated ERK1 / 2 and total ERK1 / 2 decreased, and the intracellular Ca2 + concentration decreased. Conclusion Crocin can promote the proliferation of human umbilical vein endothelial cells by activating the ERK1 / 2 signaling pathway and increasing intracellular Ca2 + concentration.