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目的探讨炎症因子白介素-6(IL-6)对人冠脉平滑肌细胞表达妊娠相关血浆蛋白A的影响。方法实验于2005年5月至2006年8月在解放军305医院老年病中心实验室完成。人冠脉平滑肌细胞37℃、5%CO2条件下常规培养。实验用5~7代的细胞,按2.5×105/mL密度接种于25cm2的培养瓶中,24h后换液,生长接近80%融合时进行如下处理:(1)应用质量浓度为10ng.mL-1的IL-6刺激人冠脉平滑肌细胞,分别在共同培养0,2,4,8,24,36h后收集细胞。(2)应用不同质量浓度的IL-6(0,5,10,50ng.mL-1)刺激人冠脉平滑肌细胞,共同培养6h后收集细胞。应用实时荧光定量聚合酶链反应的方法检测细胞内妊娠相关血浆蛋白A基因的表达量。应用SPSS10.0软件进行统计学描述与分析,所得数据用x-±s表示,多组间比较用onewayANOVA检验,两两比较应用SNK法检验,并进行pearson相关分析,P<0.05为差异有显著性意义。结果同剂量白介素-6刺激下,妊娠相关血浆蛋白A的表达量在2h时就开始发生上调,8h达高峰,而后开始下降;在不同剂量白介素-6刺激下,妊娠相关血浆蛋白A的表达量在实验剂量范围内随着白介素-6的剂量加大呈上升趋势。除5ng.mL-1和10ng.mL-1组之间差异无显著性意义(P=0.065),其余组间差异均有显著性意义(P<0.05)。结论炎症因子白介素-6能促进冠脉平滑肌细胞中斑块稳定相关标志物妊娠相关血浆蛋白A的表达,可能是炎症在急性冠脉综合征的发生发展中起重要作用的机制之一。
Objective To investigate the effect of interleukin-6 (IL-6) on the expression of pregnancy associated plasma protein A in human coronary artery smooth muscle cells. Methods The experiment was performed in the laboratory of Geriatrics Center of 305 Hospital of People’s Liberation Army from May 2005 to August 2006. Human coronary artery smooth muscle cells were cultured routinely at 37 ° C under 5% CO2. The cells were passaged at a density of 2.5 × 10 5 / mL in 25 cm 2 culture flasks for 5 to 7 days after the experiment. The medium was changed after 24 hours and the growth was approached to 80%. The following treatments were performed: (1) 1 IL-6 stimulated human coronary artery smooth muscle cells, were co-cultured 0,2,4,8,24,36 h after collecting cells. (2) Human coronary artery smooth muscle cells were stimulated with different concentrations of IL-6 (0, 5, 10 and 50 ng.mL-1) and cultured for 6 hours. Real-time fluorescence quantitative polymerase chain reaction method was used to detect the expression of intracellular pregnancy related plasma protein A gene. SPSS10.0 software was used for statistical analysis and analysis. The data was expressed as x- ± s. The multiple groups were compared by oneway ANOVA, and the pairwise comparison was performed by SNK test and pearson correlation analysis. P <0.05 was considered as significant difference Sexual meaning. Results With the same dose of interleukin-6 stimulation, the expression of pregnancy-related plasma protein A began to rise at 2h, peaked at 8h, and then began to decline. Under different doses of interleukin-6 stimulation, the expression of pregnancy-related plasma protein A In the experimental dose range with the dose of interleukin -6 increased upward trend. There was no significant difference between the 5ng.mL-1 and 10ng.mL-1 groups (P = 0.065), the other groups had significant difference (P <0.05). Conclusion Interleukin-6 (IL-6) can promote the expression of pregnancy-related plasma protein A in coronary smooth muscle cells. It may be one of the mechanisms that inflammation plays an important role in the development of acute coronary syndrome.