细胞因子对人冠脉平滑肌细胞PAPP-A、MMP-3、TIMP-1基因表达的影响  

The influence of cytokine on PAPP-A,MMP-3,TIMP-1 gene expression in human coronary artery smooth muscle cells

作　　者：夏爱祥[1] 张清华[1] 蒋知新[2] 方效民 姜同学[3] 林虎[3] 梁雪梅[3] 

机构地区：[1]解放军第305医院老年病中心,北京100017 [2]解放军第305医院中心实验室,北京100017 [3]解放军第305医院内科,北京100017

出　　处：《中国老年学杂志》2008年第11期1041-1044,共4页Chinese Journal of Gerontology

基　　金：全军医药卫生“十五”计划重点课题(04LX048);全军医药卫生“十一五”计划科技攻关课题(06G144)

摘　　要：目的探讨细胞因子白细胞介素(IL)-1β、IL-6对人冠脉平滑肌细胞中妊娠相关血浆蛋白-A(PAPP-A)、基质金属蛋白酶-3(MMP-3)、金属蛋白酶组织抑制剂-1(TIMP-1)基因表达的影响。方法应用相同浓度IL-1β(20μg/L)、IL-6(10μg/L)各自刺激人冠脉平滑肌细胞,共同培养0、2、4、8、24、36h后收集细胞。应用不同浓度IL-1β(0、5、20、40μg/L)、IL-6(0、5、10、50μg/L)刺激人冠脉平滑肌细胞,共同培养6h后收集细胞。应用实时定量PCR的方法检测细胞内PAPP-A、MMP-3、TIMP-1基因表达量。结果同剂量IL-1β刺激下,MMP-3和PAPP-A基因表达量在2 h时开始发生上调,8 h左右达高峰,而后开始下降;而TIMP-1表达量在2h时开始发生下降,8h左右达最低,而后开始上升。在不同剂量IL-1β刺激下,MMP-3、PAPP-A基因表达量随着剂量加大呈上升趋势(MMP-3:r=0.907,P=0.000;PAPP-A:r=0.972,P=0.000),TIMP-1呈下降趋势(r=-0.768,P=0.004)。不同剂量组MMP-3、TIMP-1、PAPP-A表达量具有显著性差异(MMP-3:F=24.047,P=0.000;TIMP-1:F=33.737,P=0.000;PAPP-A:F=264.699,P=0.000)。MMP-3在20μg/L和40μg/L组间没有显著性差异(P=0.154);TIMP-1仅40μg/L组和其他组间具有显著性差异,其余组间无显著性差异(P=0.383);PAPP-A在各组间均有显著性差异。在同剂量IL-6的刺激下,MMP-3、PAPP-A表达量随时间变化趋势和IL-1β相似,而TIMP-1则在4 h时就达到最低,随后开始上升。在不同剂量IL-6刺激下,MMP-3、PAPP-A亦有随剂量加大表达量上升趋势(MMP-3:r=0.919,P=0.000;PAPP-A:r=0.941,P=0.000),TIMP-1呈下降趋势(r=-0.799,P=0.002)。不同剂量组MMP-3、TIMP-1、PAPP-A表达量均有显著性差异(MMP-3:F=14.081,P=0.001;TIMP-1:F=5.727,P=0.022;PAPP-A:F=25.128,P=0.000)。MMP-3在5μg/L和10μg/L组间没有显著性差异(P=0.292);TIMP-1对照组与10μg/L和50μg/L组间具有显著性差异,其余各组间没有显著性差异(P=0.253);PAPP-A基因表达量在5μg/L和10μg/L组没有显著性差异(P=0.065)。结论炎症因Objective To explore the effect of IL-1β and IL-6 on pregnancy-associated plasma protein （PAPP）-A, MMP-3, TIMP- 1 gene expression in human coronary artery smooth muscle cells. Methods Human coronary artery smooth muscle cells were stimulated with IL-1β （ 20 μg/L） or IL-6 （ 10 μg/L） to isolate RNA and collect culture media after 0, 2, 4, 8, 24 and 36 h. Then human coronary artery smooth muscle cells were stimulated with IL-1β （0, 5, 20, 40 μg/L） or IL-6（0, 5, 10, 50 μg/L） respectively to isolate RNA and collect culture media after 6 h. Real-time PCR was performed on RNA to detect gene expression. Results In the same concentration of IL-1β, MMP-3, PAPP-A mRNA expressions were up-regulated at 2 hour, at 8 hour the expression reached the peak, then began to descend; TIMP- 1 mRNA expression was down-regulated at 2 hour,the mRNA expression got to the lowest at 8 hour, then began to go up. In different concentration of IL-1β, MMP-3, PAPP-A mRNA expression was up-regulated with the dose （ MMP-3 ： r = 0. 907, P = 0. 000 ; PAPP-A ： r = 0. 972, P =0. 000） , and TIMP-1 mRNA expression was down-regnlated with the dose （r = -0. 768,P =0. 004）. There were significant difference in mRNA expression among different groups （ MMP-3 ： F = 24. 047, P = 0. 000 ; TIMP-1 ： F = 33. 737, P = 0. 000 ; PAPP-A ： F = 264. 699, P = 0. 000）. There were no significant differences of MMP-3 between 20 and 40 μg/L groups （P =0. 154）. But there was significant difference of TIMP-1 mRNA expression only between 40 μg,/L and other groups ; In the same concentration of IL-6, MMP-3, PAPP-A mRNA expression was upegulated at 2 hour, at 8 hour the expression reached peak, then began to descend; TIMP-1 mRNA expression was down-regulated at 2 hour, the mRNA expression got to the lowest at 4 hour, then began to increase. In different concentration of IL-6, MMP-3, PAPP-A mRNA expression was up-regnlated with the dose（MMP-3 ：r =0. 919,P =0. 000;PAPP-A：r =0. 941 ,P =0. 000）, and TIMP-1 mRNA expressio

关 键 词：急性冠脉综合征 白细胞介素-1β 白细胞介素-6 妊娠相关血浆蛋白-A 基质金属蛋白酶-3 基质金属蛋白组织抑制剂-1 

分 类 号：R541.4[医药卫生—心血管疾病]