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目的:观察搏动信号对人脐静脉内皮细胞中反映内皮细胞活性的内皮型一氧化氮合酶mRNA表达的影响。方法:实验于2004-06/2005-03在湘雅二医院心胸外科生物材料研究室和上海生物工程有限公司进行。将取自剖宫产产妇自愿捐献的无菌脐带的人脐静脉内皮细胞培养、传代,传4代后分为两组:①搏动培养组:先静放6～8h待细胞贴壁后,用自行设计的细胞搏动培养装置,进行搏动培养。搏动参数:搏动频率150次/min,搏出量0.3mL/次,搏动产生的压力30/9mmHg(1mmHg=0.133kPa)。②静态培养组:不进行搏动,其余条件同搏动培养组。每组每个时间点5个样本。用反转录-聚合酶链反应方法,分别在第1,2,4,6,9,12,15,18天检测两组的人脐静脉内皮细胞的内皮型一氧化氮合酶mRNA的表达水平(相对吸光度值)。结果:人脐静脉内皮细胞的内皮型一氧化氮合酶mRNA表达:①培养第1,4天,静态培养组明显高于搏动培养组(1.0±0.55比1.10±0.35;1.17±0.23比1.42±0.44,P<0.05)。②培养第2天两组比较差异不显著(1.16±0.18比1.25±0.35,P>0.05)。③培养第6,9,12,15,18天,搏动培养组明显高于静态培养组(1.52±0.14比1.27±0.30;1.60±0.19比1.16±0.33;1.68±0.44比0.99±0.31;1.7±0.24比1.1±0.94;1.7±0.16比1.0±0.28,P<0.05)。结论:搏动信号能提高人脐静脉内皮细胞的内皮型一氧化氮合酶的表达。提示搏动信号在维持细胞发挥正常的生理功能方面起到重要的作用,能增强细胞的生物活性。
Objective: To observe the effect of beating signal on the expression of endothelial nitric oxide synthase mRNA in human umbilical vein endothelial cells. METHODS: The experiment was performed at the Department of Cardiothoracic Surgery, Shanghai Second Xiangya Hospital from June 2004 to March 2005 with Shanghai Bioengineering Co., Ltd. Cultured umbilical vein endothelial cells from sterile umbilical cord donated by cesarean mothers voluntarily were passaged and passaged for 4 generations and then divided into two groups: ① beating culture group: first resting for 6 ~ 8 hours, Self-designed cell pulsation device for pulsatile culture. Beating parameters: pulsatile frequency 150 beats / min, stroke volume 0.3mL / time, beating the pressure generated 30 / 9mmHg (1mmHg = 0.133kPa). ② static culture group: no pulsation, the remaining conditions with pulsatile culture group. Each group of five samples at each time point. The expression of endothelial nitric oxide synthase mRNA in human umbilical vein endothelial cells was detected on the 1st, 2nd, 4th, 6th, 9th, 12th, 15th and 18th day by reverse transcription polymerase chain reaction Horizontal (relative absorbance value). Results: The expression of endothelial nitric oxide synthase mRNA in human umbilical vein endothelial cells: ①In the first and the fourth day of culture, the static culture group was significantly higher than the pulsatile culture group (1.0 ± 0.55 vs 1.10 ± 0.35; 1.17 ± 0.23 vs 1.42 ± 0.44, P <0.05). ② On the second day, there was no significant difference between the two groups (1.16 ± 0.18 vs 1.25 ± 0.35, P> 0.05). ③ On the 6th, 9th, 12th, 15th and 18th day, the pulsatile culture group was significantly higher than the static culture group (1.52 ± 0.14 vs 1.27 ± 0.30; 1.60 ± 0.19 vs 1.16 ± 0.33; 1.68 ± 0.44 vs 0.99 ± 0.31; 1.7 ± 0.24 vs. 1.1 ± 0.94; 1.7 ± 0.16 vs. 1.0 ± 0.28, P <0.05). Conclusion: Beating signals can increase the expression of endothelial nitric oxide synthase in human umbilical vein endothelial cells. It is suggested that the beating signal plays an important role in maintaining the normal physiological functions of cells and enhancing the biological activity of the cells.