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目的:探讨孟鲁司特对人单核细胞株(THP-1)源泡沫细胞的形成及基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)蛋白表达和分泌的影响,为该药防治动脉粥样硬化提供实验依据。方法:用佛波酯(phorbol myristate acetate,PMA)诱导THP-1分化为巨噬细胞,然后给予氧化型低密度脂蛋白(oxidized low density liprotein,OX-LDL)孵育48 h,建立泡沫细胞模型。以不同浓度的孟鲁司特(0.1,0.01μmol·L-1)及MMP-2/9抑制剂(20μmol·L-1)预处理巨噬细胞4 h后再加入OX-LDL孵育48 h。油红O染色观察泡沫细胞的形成,并通过Western blot和明胶酶谱Gelatin zy-mography方法分别检测细胞中MMP-2/9蛋白的表达及上清中MMP-2/9的活性。结果:巨噬细胞未被油红O染色,泡沫细胞组大量细胞胞质被油红O染成红色,而孟鲁司特及MMP-2/9抑制剂处理组细胞胞质染色明显浅于泡沫细胞组。THP-1细胞向巨噬细胞、泡沫细胞分化过程中,细胞中MMP-2/9蛋白的表达(P<0.05)和上清中MMP-2/9的活性均逐步增加(P<0.05);而孟鲁司特显著下调泡沫细胞中MMP-2/9蛋白的表达(P<0.05)和上清中MMP-2/9的活性(P<0.05)。结论:孟鲁司特可抑制泡沫细胞的形成及MMP-2/9的生成,为防治动脉粥样硬化及抗动脉粥样硬化新药的筛选提供新的理论依据。
Objective: To investigate the effects of montelukast on the formation of THP-1-derived foam cells and the expression of matrix metalloproteinase-2 (MMP-2), matrix metalloproteinase-9 , MMP-9) protein expression and secretion of the drug for the prevention and treatment of atherosclerosis provide experimental evidence. METHODS: THP-1 cells were induced to differentiate into macrophages with phorbol myristate acetate (PMA), then incubated with oxidized low density lipoprotein (OX-LDL) for 48 h to establish a foam cell model. Macrophages were pretreated with different concentrations of montelukast (0.1, 0.01 μmol·L-1) and MMP-2/9 inhibitor (20 μmol·L-1) for 4 h and then incubated with OX-LDL for 48 h. The formation of foam cells was observed by oil red O staining. The expression of MMP-2/9 protein and the activity of MMP-2/9 in the supernatant were detected by Western blot and gelatin zy-mography. RESULTS: Macrophages were not stained with Oil Red O, and a large number of cytoplasm of foam cells were stained red with Oil Red O. Cytoplasmic staining of cells treated with montelukast and MMP-2/9 inhibitors was significantly lighter than that of foam Cell group. The expression of MMP-2/9 (P <0.05) and the activity of MMP-2/9 in the supernatant of THP-1 cells gradually increased (P <0.05) during the differentiation of the macrophages and foam cells. While montelukast significantly downregulated the expression of MMP-2/9 protein (P <0.05) and the activity of MMP-2/9 (P <0.05) in the supernatant. Conclusion: Montelukast can inhibit the formation of foam cells and the production of MMP-2/9, providing a new theoretical basis for the prevention and treatment of atherosclerosis and screening of new anti-atherosclerotic drugs.