摘要
目的探讨miRNA-143在同型半胱氨酸(homocysteine,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。
Aim To explore the effect of miRNA-143 （miR-143 ） on homocysteine （ Hey ） induced - vascular smooth muscle cells （ VSMCs ） proliferation and themechanism. Methods VSMCs were cultured and incubatedwith Hey by using primary cultured method.T h e n, cells were treated with different concentrationsof Hey and folate. VSMCs proliferation was determinedwith M T T assay,miR -143 was measured by qRT-PCR ,and methylation of miR -143 was determined with methylatedP CR . Results After cells were treated with differentconcentrations of Hey , the proliferation ofVSMCs was significantly increased, mRNA expressionof miR -143 was decreased and methylation of miR -143was increased. The proliferation of VSMCs was significantlydecreased when transfected VSMCs with miR -143 precursor,and cell proliferation was increased byusing miR -143 inhibitor transfection. Conclusion H y -pomethylation of miR -143 may inhibit VSMCs proliferation.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2016年第8期1097-1101,共5页
Chinese Pharmacological Bulletin
基金
宁夏自然科学基金资助项目(No NZ15209)
国家自然科学基金资助项目(No 81360027
81360052
81360053)
