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-  2016 

苏子油干预血管内皮细胞miR-21靶向调控MMP-9的机制
Mechanism of Perilla oil’s intervention in vascular endothelial cell miR-21 targeted regulation of MMP-9

DOI: 10.7652/jdyxb201601024

Keywords: 苏子油,微小RNA-21,基质金属蛋白酶-9,靶向调控,抗动脉粥样硬化
Perilla oil
,miR-21,MMP-9,targeted control,anti-atherosclerosis

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Abstract:

摘要:目的 探讨不同浓度苏子油干预人脐静脉血管内皮细胞(HUVEC) miR-21和基质金属蛋白酶MMP-9的表达,为苏子油改善动脉粥样硬化提供依据。方法 体外培养HUVEC,以TNF-α诱导细胞进行造模,将细胞分为空白组、模型组及苏子油高、中、低剂量组。MMT法检测各组细胞A值;PT-PCR检测细胞miR-21、MMP-9基因水平;脂质体转染法将miR-21模拟物转入HUVEC细胞,Western blot检测转染后MMP-9蛋白的表达。结果 苏子油高、中、低剂量能明显改善TNF-α诱导的HUVEC损伤,且苏子油对内皮细胞的保护作用呈剂量依赖关系;RT-PCR结果显示,模型组miR-21及MMP-9的表达高,苏子油干预后,miR-21及MMP-9基因的表达逐渐减弱,且二者分别与模型组比较,差异有统计学意义(P<0.05);细胞转染miR-21后,MMP-9蛋白的表达显著增强;miR-21与MMP-9 mRNA 3′UTR区经生物信息学软件预测存在靶向结合位点。结论 苏子油可能通过干预miR-21靶向调控MMP-9的表达对血管内皮细胞发挥保护作用,从而发挥抗动脉粥样硬化作用。
ABSTRACT: Objective To explore the intervention of different concentrations of Perilla oil in miR-21 and matrix metalloproteinase 9 (MMP-9) expressions of human umbilical vein endothelial cells (HUVECs) to provide the basis for Perilla oil??s improvement of atherosclerosis. Methods In vitro umbilical vein endothelial cells were induced by TNF-α, and then divided into blank group, model group as well as Perilla oil high-, medium- and low-dose groups. A value was detected by MMT method, and gene levels of cell miR-21 and MMP-9 were detected by PT-PCR. Liposome transfection method was used to put the miR-21 analog into HUVEC cells; the expression of MMP-9 protein after transfection was detected by Western blot. Results Perilla oil of three doses could obviously improve HUVEC injury induced by TNF-α, and Perilla oil??s protection of endothelial cells was dose-dependent. miR-21 and MMP-9 gene expressions detected by RT-PCR showed that both of them were higher in model group. After Perilla oil intervention, the expression of miR-21 and MMP-9 gene gradually decreased with the dose of Perilla oil, both of which differed significantly from those in model group (P<0.05). After cell transfection of miR-21, the expression MMP-9 protein was significantly increased. MiR-21 and MMP-9 mRNA 3??UTR region detected by bioinformatics software could predict target binding site. Conclusion Perilla oil can protect vascular endothelial cells by intervening in miR-21??s regulation of MMP-9 expression, thus having anti-atherosclerosis effect

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