摘要
目的:探讨血管紧张素(Ang)Ⅱ在诱导巨噬细胞表达细胞外基质金属蛋白酶诱导因子(EMMPRIN)中的作用及机制。方法:以5ng/ml佛波醇诱导人类单核细胞株细胞成巨噬细胞后,倒置相差显微镜观察诱导情况;四甲基偶氮唑盐微量酶反应比色法监测不同AngⅡ刺激下的细胞活性;逆转录聚合酶链式反应及蛋白质印迹检测不同时间及不同浓度的AngⅡ对EMMPRIN基因及蛋白表达的影响;进一步用AngⅠ受体拮抗剂(10μM)及AngⅡ受体拮抗剂(10μM)探讨信号传导机制,检测AngⅡ对EMMPRIN的诱导情况。结果:在5ng/ml佛波醇诱导下可很好的建立巨噬细胞模型。四甲基偶氮唑盐微量酶反应比色法结果示AngⅡ在0.01～10μM浓度下,细胞的活性不受影响(P>0.05),50μMAngⅡ刺激下细胞活性改变(P<0.05),差异有统计学意义。不同浓度AngⅡ刺激下巨噬细胞内EMMPRIN基因及蛋白的表达成时间依赖性,6h后开始增高(P<0.05),12h后达峰值(P<0.01),24h后开始下降(P<0.05),差异有统计学意义;AngⅡ对EMMPRIN的刺激作用亦呈剂量依赖型,随着浓度(0μM、0.01μM、0.1μM和1μM)的增加,EMMPRIN基因及蛋白的表达亦明显增加(P<0.05),差异有统计学意义;进一步的信号通路研究实验结果示AngⅠ受体拮抗剂可抑制AngⅡ的诱导作用(P<0.05),AngⅡ受体拮抗剂则无影响。结论:在佛波醇诱导的人类单核细胞株细胞中,AngⅡ通过AngⅠ受体上调巨噬细胞中EMMPRIN的表达,氯沙坦可抑制其上调作用。
Objective： To explore angiotensin Ⅱ （Ang Ⅱ） induced human extra cellular matrix metaUoproteinase inducer （EMMPRIN） expression in macrophages with its underlying mechanisms. Methods： The macrophage model was established by 5ng/ml phorbol 12-myristate 13-acetate （PMA） induction on human monocyte cell line THP-1, the cells were observed with inverted phase contrast microscope. Cyto-activity stimulated by different concentrations of Ang Ⅱ was measured by methy thiazolyl tetrazolium（MTT） enzyme method. EMMPRIN gene and its protein expression were examined by RT-PCR and Western blot analysis. In order to further study the signal transduction mechanism of Ang Ⅱ ,receptor antagonist AT, （10 μM） and antagonist AT2 （10 μM） were used to inhibit the effect of AngⅡ and to observe the expression change of EMMPRIN. Results ： MTT enzyme method demonstrated that cyto-activity was not influenced by stimulation of 0. 01 μM-10 μM Ang Ⅱ（P 〉 0. 05 ）, but cyto-activity was changed by stimulation of 50 μM Ang Ⅱ （ P 〈 0. 05 ）. The stimulation effect of Ang Ⅱwas time-dependent, EMMPRIN gene and its protein expression began to increase after 6 h （P 〈 0. 05 ）of stimulation, peaked at 12 h （P 〈 0. 01 ）,and declined after 24 h（P 〈0. 05）. The stimulation effect of Ang Ⅱ was also dose-dependent, with the elevation of Ang Ⅱ concentration ,the expression of EMMPRIN was increased（P 〈0. 05 ）. Further signal transduction study demonstrated that ATI -receptor antagonist could inhibit the stimulation effect of Ang Ⅱ（P 〈0. 05） but AT2-receptor antagonist could not do that（P 〉0. 05）. Conclusion： Ang Ⅱ up-regulated EMMPRIN expression in macrophages, and this effect could be inhibited by Losartan.
出处
《中国循环杂志》
CSCD
北大核心
2010年第1期58-61,共4页
Chinese Circulation Journal
基金
云南省社会发展科技计划(2008C150M)
