miRNA－143在同型半胱氨酸致血管平滑肌细胞增殖中作用的研究

摘要

Aim To explore the effect of miRNA-143 ( miR-1 4 3 ) on homocysteine ( Hcy ) induced-vascular smooth muscle cells ( VSMCs ) proliferation and the mechanism .Methods VSMCs were cultured and in-cubated with Hcy by using primary cultured method . Then, cells were treated with different concentrations of Hcy and folate .VSMCs proliferation was determined with MTT assay , miR-143 was measured by qRT-PCR, and methylation of miR-143 was determined with meth-ylated PCR.Results After cells were treated with dif-ferent concentrations of Hcy , the proliferation of VSMCs was significantly increased , mRNA expression of miR-143 was decreased and methylation of miR-143 was increased .The proliferation of VSMCs was signifi-cantly decreased when transfected VSMCs with miR-143 precursor , and cell proliferation was increased by using miR-143 inhibitor transfection .Conclusion Hy-pomethylation of miR-143 may inhibit VSMCs prolifera-tion.%目的：探讨 miRNA－143在同型半胱氨酸（ homocys－teine， Hcy）致血管平滑肌细胞增殖中的作用。方法原代培养血管平滑肌细胞（ vascular smooth muscle cells ， VSMCs），用不同浓度的 Hcy 和叶酸干预， MTT 法检测VSMCs增殖情况，qRT－PCR法检测miR－143的表达，巢式降落式特异性甲基化PCR法分析miR－143启动子区的甲基化状态，分别用miR－143的前体和抑制物转染细胞检测细胞增殖。结果不同浓度的Hcy均可以引起VSMCs增殖活性增强，miR－143的mRNA表达降低，miR－143启动子区甲基化程度升高，以100μmol・ L－1 Hcy最为明显（ P＜0．01 ）， miR－143前体作用VSMCs后，细胞的增殖活性减弱（ P＜0．01 ），细胞转染miR－143的抑制物后细胞增殖活性明显增强（ P＜0．01）。结论 miR－143可以抑制VSMCs增殖，其机制可能与miR－143启动子区高甲基化有关。