|
- 2016
苏子油干预血管内皮细胞miR-21靶向调控MMP-9的机制
|
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
[1] | 李毅,孙瑞红,肖玲,等. MMP-9及NF-κB对人颅内动脉粥样硬化斑块稳定性的影响[J]. 中风与神经疾病杂志, 2009, 26(4):396-398. |
[2] | 张天. MMP-9、TIMP-1、CD147在人粥样硬化冠状动脉中表达的相关性研究[D]. 大连医科大学,2012. |
[3] | 张政. MicroRNA靶向PTEN/PI3K信号途径调控早期糖尿病肾病的研究[D]. 重庆医科大学,2009. |
[4] | 李英霞,张岩. 苏子油复方制剂对高脂血症模型大鼠血栓素B2和6-酮-前列腺素F1A的影响[J]. 中医药导报, 2007, 13(9):17-18. |
[5] | 袁梅. MMP-9基因3??UTR多态性与动脉粥样硬化性脑梗死相关性及miRNA-491介导的多态调控机制研究[D]. 中南大学湘雅学院,2012. |
[6] | 于晓红,王小明,李瑶,等. NADHP氧化酶在TNF-α诱导HUVEC HO-1表达中的作用[J]. 基础医学与临床,2006, 26(5):461-465. |
[7] | 姚义安,张抒扬. 内皮细胞与动脉粥样硬化[J]. 中华内科杂志,2008, (1):63-64. |
[8] | 鲁其良,毕立志,乔涛. 阿托伐他汀对急性冠脉综合征患者血清基质金属蛋白酶-9及其组织抑制因子-1的影响[J]. 中国基层医药, 2010, 17(19):2653-2654. |
[9] | 杨春海,代全德,张建平,等. 依那普利对肾性高血压大鼠脑缺血再灌注后脑组织基质金属蛋白酶-2、-9表达的影响[J]. 中国基层医药, 2010, 17(15):2081-2083. |
[10] | 周文阳. 三七皂苷对内皮细胞MMP-活性、巨噬细胞炎症因子表达及血小板凝集抑制作用的实验研究[D]. 中国医科大学, 2012. |
[11] | 陈君君. MicroRNA-21在ox-LDL诱导的血管内皮细胞中的功能及丹皮酚的干预作用[D]. 安徽中医药大学,2013. |
[12] | GHODKE Y, CHOPRA A, SHINTRE P, et al. Profiling single nucleotide polymorphisms (SNPs) across intracellular folate metabolic pathway in healthy Indians[J]. Indian J Med Res, 2011, 133(3):274-279. |
[13] | 熊雁. 血管内皮细胞microRNA-21在高糖环境及冠心病病人中表达的差异及其功能研究[D]. 遵义医学院, 2012. |
[14] | 赵峰. miR-335与胃癌侵袭转移的相关性及其机制的研究[D]. 中国医科大学,2012. |
[15] | 孙恕. 他汀类干预血管内皮细胞基质金属蛋白酶-1的表达[D]. 江西医学院,2004. |
[16] | 王子栋. miR-21在大鼠血管平滑肌细胞中对MMP2、MMP9表达的影响[D]. 哈尔滨医科大学,2012. |