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目的探讨不同浓度转化生长因子(TGF)-β_1对人主动脉平滑肌细胞(HA-VSMCs)增殖及迁移的影响。方法体外培养人主动脉平滑肌细胞,用0、0.05、0.1、0.5、1、2、5、10ng/ml浓度的重组人TGF-β_1进行干预,分别在干预后第0、6、12、24、36小时5个不同时间点进行CCK-8试验,或24小时后进行Transwell迁移试验,观察细胞增殖及迁移情况。结果当TGF-β_1浓度<5ng/ml时,随着刺激浓度增加,人主动脉VSMCs增殖水平逐渐升高,当TGF-β_1浓度>5 ng/ml时,细胞增殖水平降低。不同浓度TGF-β_1(0、0.05、0.1、0.5、1、2、5、10 ng/ml)干预细胞,随着作用时间延长,细胞增殖水平逐渐升高,在24小时时达到高峰,之后逐渐降低。TGF-β_1浓度为5 ng/ml且培养时间为24小时时细胞增殖水平最高(OD值:0.843±0.016)(P<0.01)。当TGF-β_1浓度≤5ng/ml(0、0.05、0.1、0.5、1、2、5 ng/ml)且作用时间为24小时时,随着TGF-β_1浓度增加,人主动脉VSMCs迁移水平不断升高,其中以TGF-β_1浓度为5 ng/ml时细胞迁移水平最高,迁移细胞数为84.667±0.577;TGF-β_1浓度>5 ng/ml时,细胞迁移水平降低,迁移细胞数为69.667±1.528。与其余各组TGF-β_1浓度作用下的迁移细胞数比较,差异有统计学意义(P<0.01)。结论 5ng/ml的TGF-β_1作用24小时更能刺激人主动脉平滑肌细胞发生增殖及迁移。
Objective To investigate the effects of different concentrations of transforming growth factor-β 1 on the proliferation and migration of human aortic smooth muscle cells (HA-VSMCs). Methods Human aortic smooth muscle cells were cultured in vitro and were treated with recombinant human TGF-β 1 at the concentrations of 0, 0.05, 0.1, 0.5, 1, 2, 5 and 10ng / ml. CCK-8 assay was performed at 36 hours and 5 different time points, or Transwell migration assay was performed 24 hours later to observe cell proliferation and migration. Results When the concentration of TGF-β 1 was lower than 5ng / ml, the proliferation of VSMCs increased gradually with the increase of stimulation concentration. When the concentration of TGF-β_1 was> 5ng / ml, the proliferation of VSMCs was decreased. Different concentrations of TGF-β 1 (0,0.05,0.1,0.5,1,2,5,10 ng / ml) intervention cells, with the extension of time, cell proliferation increased gradually reached its peak at 24 hours, and then gradually reduce. TGF-β 1 concentration of 5 ng / ml and cultured for 24 hours, the highest level of cell proliferation (OD value: 0.843 ± 0.016) (P <0.01). When the concentration of TGF-β 1 was lower than 5ng / ml (0,0.05,0.1,0.5,1,2,5 ng / ml) and the action time was 24 hours, the migration of VSMCs in the aorta continued to increase with the increase of TGF- The highest level of cell migration was obtained when the concentration of TGF-β 1 was 5 ng / ml, the number of migrating cells was 84.667 ± 0.577. When the concentration of TGF-β 1 was above 5 ng / ml, the cell migration was decreased and the number of migrating cells was 69.667 ± 1.528. Compared with other groups, the number of migrating cells under the action of TGF-β 1 was significantly different (P <0.01). Conclusion TGF-β 1 at 5ng / ml can stimulate the proliferation and migration of human aortic smooth muscle cells for 24 hours.