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目的探讨纤维蛋白诱导的自噬对内皮细胞增殖和血管腔样结构形成的作用。方法 MTT法观察在不同浓度和时间的纤维蛋白作用下,人脐静脉内皮细胞(HUVECs)形成血管腔样结构和细胞增殖的改变;采用RT-PCR检测不同浓度(1.5、3.0和6.0mg/ml)和时间(6、12和24h)的纤维蛋白作用下,微管相关蛋白1轻链3(MAP1LC3)和血管内皮生长因子(VEGF)的表达。用自噬抑制剂3-甲基腺嘌呤(3-MA)干预作用最显著的纤维蛋白浓度和时间,观察细胞增殖、血管形成和基因表达变化。结果 HUVECs在纤维蛋白作用下,可见管腔样结构形成,并呈浓度和时间依赖性;细胞增殖也较对照组明显增加。LC3和VEGF的表达均随着纤维蛋白浓度和时间的增加而增加。但在自噬抑制剂3-MA的作用下,细胞增殖降低,未见明显的新生血管形成;并且LC3、VEGF的mRNA表达也明显减少。结论纤维蛋白激活自噬,参与了细胞增殖和血管新生的过程。
Objective To investigate the effect of fibrin-induced autophagy on endothelial cell proliferation and vascular cavity-like structure formation. Methods MTT assay was used to observe the changes of vascular cavity-like structure and cell proliferation in human umbilical vein endothelial cells (HUVECs) under the action of different concentrations and time of fibrin. ) And time (6, 12 and 24h) fibrin, microtubule-associated protein 1 light chain 3 (MAP1LC3) and vascular endothelial growth factor (VEGF) expression. The autophagy inhibitor 3-methyladenine (3-MA) was used to intervene in the most significant fibrin concentration and time, and the cell proliferation, angiogenesis and gene expression were observed. Results Under the action of fibrin, HUVECs showed lumen-like structure formation in a concentration- and time-dependent manner. The proliferation of HUVECs was also significantly increased compared with that of the control group. Both LC3 and VEGF expression increased with increasing fibrin concentration and time. However, under the action of autophagy inhibitor 3-MA, cell proliferation decreased, no obvious neovascularization was found, and mRNA expressions of LC3 and VEGF were also significantly decreased. Conclusion Fibrin activation of autophagy, involved in cell proliferation and angiogenesis.