| 192 | 8 | 63 |
| 下载次数 | 被引频次 | 阅读次数 |
为探究mi R-181b-5p靶向调控TIMP3对apo E-/-鼠动脉粥样硬化炎症的影响,本研究基于生物信息学预测筛选,并采用双荧光素酶报告基因系统验证mi R-181b-5p靶基因。同时在ox-LDL刺激THP-1巨噬细胞及apo E-/-动脉粥样硬化模型小鼠中检测mi R-181b-5p靶基因m RNA及蛋白水平的表达情况,并且检测转染mi R-181b-5p后细胞水平和动物体内炎症因子TNF-α、INF-γ、IL-1β、IL-18、CCL2的表达变化。此外,通过病理切片方法检测转染mi R-181b-5p后小鼠主动脉窦处病变情况。研究通过双荧光素酶报告基因系统验证TIMP3为mi R-181b-5p靶基因,并在ox-LDL刺激THP-1巨噬细胞及apo E-/-动脉粥样硬化模型小鼠中证明了mi R-181b-5p可以降低TIMP3转录水平从而降低其蛋白水平表达,同时研究发现mi R-181b-5p在细胞水平和动物体内均会增强炎症因子TNF-α、INF-γ、IL-1β、IL-18、CCL2的表达,并可以增加小鼠主动脉窦处病变面积。研究认为在动脉粥样硬化发生中,mi R-181b-5p可以增加病变面积并且提升相关炎症因子的表达、分泌,其作用机理很可能是通过抑制靶基因TIMP3表达,进而降低TIMP3蛋白水平实现的,为研究动脉粥样硬化机制提供了理论依据。
Abstract：To explore effect of miR-181 b-5 p targeted regulating on TIMP3 in apo E-/-arterial atherosclerosis inflammation,the target gene of mi R-181 b-5 p was validated by bioinformatics prediction screening,double luciferase reporter gene system was applied.After that,mi R-181 b-5 p target gene m RNA and protein expression levels were detected in ox-LDL-stimulated THP-1 macrophages and apo E-/-atherosclerotic mice model.The expression variation of TNF-α,INF-γ,IL-1β,IL-18 and CCL2 in cellular level and in vivo were detected after mi R-181 b-5 p transfection.What's more,the lesion in aortic sinus was detected by pathological section after mi R-181 b-5 p transfection.The dual-luciferase reporter system verified mi R-181 b-5 p target gene-TIMP3.It was also demonstrated that mi R-181 b-5 p could degrade TIMP3 mRNA and then reduce protein expression level in ox-LDL-stimulated THP-1 macrophages and apo E-/-atherosclerotic mice model.miR-181 b-5 p can enhance the expression of TNF-α,INF-γ,IL-1β,IL-18 and CCL2 in the cellular level and in vivo.In addition,mi R-181 b-5 p can enhance the lesion area in mice aortic sinus significantly.In the development of atherosclerosis,mi R-181 b-5 p can increase the lesion area and enhance the secretion of related inflammatory factors.The functional mechanism of mi R-181 b-5 p is likely to be TIMP3 m RNA degradation and TIMP3 protein reduction,which provide theoretical basis for atherosclerosis.
Bhela S.,Mulik S.,Gimenez F.,Reddy P.B.J.,Richardson R.L.,Varanasi S.K.,Jaggi U.,Xu J.,Lu P.Y.,and Rouse B.T.,2015,Role of mi R-155 in the pathogenesis of herpetic stromal keratitis,Am.J.Pathol.,185(4):1073-1084
Dieter B.P.,and Tuttle K.R.,2016,Dietary strategies for cardiovascular health,Trends.Cardiovasc.Med.,12(7):1-16
Fan J.Y.,Mi X.N.,Chen Y.,He S.J.,Wang L.,Hui R.T.,and Zhang W.L.,2014,Regulatory role of micro RNA-181b in the angiogensis of senescent endothelial cells,Zhongguo Fenzi Xinzangbingxue Zazhi(Molecular Cardiology of China),14(1):826-829(范婧尧,米雪楠,陈宇,贺淑君,王雷,惠汝太,张伟丽,2014,Micro RNA-181b对衰老内皮细胞血管新生功能的影响,中国分子心脏病学杂志,14(1):826-829)
Giral H.,Kratzer A.,and Landmesser U.,2016,micro RNAs in lipid metabolism and atherosclerosis,Best Practice Research Clinical Endocrinology Metabolism,30:665-676
Karasawa T.,and Takahashi M.,2017,Role of NLRP3 inflammasomes in atherosclerosis,J.Atheroscler.Thromb.,24:1-9
Laffont B.,and Rayner K.J.,2017,Micro RNAs in the pathobiology and therapy of atherosclerosis,Canadian Journal of Cardiology,33(3):313-324
Lee Y.T.,Lin H.Y.,Chan Y.W.F.,Li K.H.C.,To O.T.L.,Yan B.P.,Liu T.,Li G.P.,Wong W.T.,Keung W.,and Tse G.,2017,Mouse models of atherosclerosis:a historical perspective and recent advances,Lipids.Health Dis.,16(1):1-11
Long G.W.,Wang F.,Li H.P.,Yin Z.W.,Sandip C.,Lou Y.,Wang Y.,Chen C.,and Wang D.W.,2013,Circulating mi R-30a,mi R-126 and let-7b as biomarker for ischemic stroke in humans,BMC Neurology,13(1):1-10
Luo Y.,Xiong W.,Yang C.Y.,and Liang X.,2015,The correlation between mi RNA-126 and plaque stability in the patients with carotid atherosclerosis,Zhongfeng Yu Shenjing Jibing Zazhi(Journal of Apoplexy and Nervous Diseases),32(8):705-707(骆瑜,熊玮,杨纯玉,梁旭,2015,mi R-NA-126与颈动脉粥样硬化患者斑块稳定性的关系,中风与神经疾病杂志,32(8):705-707)
Oguz E.G.,Akoglu H.,Okyay G.U.,Gursoy G.K.,Yildirim T.,Merhametsiz O.,Cimen T.,Canbakan B.,Yeter E.,and Ayli M.D.,2017,Increased serum renalase in peritoneal dialysis patients:Is it related to cardiovascular disease risk?Nefrologia,314:1-6
Psaltopoulou T.,Hatzis G.,Papageorgiou N.,Androulakis E.,Briasoulis A.,and Tousoulis D.,2017,Socioeconomic status and risk factors for cardiovascular disease:impact of dietary mediators,Hellenic J.Cardiol.,132:1-27
Shao J.L.,Li Z.Z.,Wang L.,Jiao G.L.,Zhou Z.G.,and Sun G.D.,2016,micro RNA-181b promotes migration and invasion of osteosarcoma cells by targeting N-myc downstream regulated gene 2,Journal of Southern Medical University,36(3):321-326
Stefania L.M.,Raluca I.M.,Carmen M.M.,Mircea C.,and Dragos V.,2015,Atherosclerosis in systemic sclerosis:a modern controversy,Maedica,10(3):248-256
Tian H.,Huang M.L.,Liu K.Y.,Jia Z.B.,Sun L.,Jiang S.L.,Liu W.,Mc Donald Kinkaid H.Y.,Wu J.,and Li R.K.,2012,Inhibiting matrix metalloproteinase by cell-based timp-3 gene transfer effectively treats acute and chronic ischemic cardiomyopathy,Cell Transplant.,21(5):1039-1053
Yang J.,Fan Z.X.,Yang J.,Ding J.W.,Yang C.J.,and Chen L.H.,2016,micro RNA-22 attenuates myocardial is chemia-reperfusion injury via an anti-inflammatory mechanism in rats,Exp.Ther.Med.,12(5):3249-3255
Zhang H.N.,2014,Increased expression of mi RNA 142-3p in the oxidative process of endothelial cells in atherosclerosis,Thesis for M.S.,Dalian Medical University,Supervisor:Liu S.M.,pp.1-30(张红娜,2014,mi RNA 142-3P在内皮细胞氧化导致动脉粥样硬化过程中的表达,硕士学位论文,大连医科大学,导师:刘双梅,pp.1-30)
Zhong W.,Yang J.,Cao Q.,and Huan X.,2015,Association between mi R-181b and PKG 1 in myocardial hypertrophy and its clinical implications,Exp.Ther.Med.,10(3):857-862
基本信息:
DOI：10.13417/j.gab.037.002200
中图分类号:R543.5
引用信息:
[1]郑莉,谭成宇.miR-181b-5p调控TIMP3对apoE~(-/-)鼠动脉粥样硬化炎症影响及相关因子检验[J].基因组学与应用生物学,2018,37(05):2200-2206.DOI:10.13417/j.gab.037.002200.
基金信息:
国家863项目(2014AA022301)资助
2018-05-25
2018-05-25