张营,吴晟,鲍海龙,刘兴德
ZHANG Ying,WU Cheng,BAO Hailong,LIU Xingde

• 1:贵州医科大学附院心内科
• 2:贵州中医药大学

摘要(Abstract)：

目的:探讨前蛋白转化酶枯草溶菌素9(PCSK9)基因5端非编码区(5′UTR)rs45448095基因多态性与血浆低密度脂蛋白(LDL)水平及冠心病风险的相关性。方法:使用千人基因组计划数据库分析中国汉族人群PCSK9基因5′UTR区基因多态性,利用荧光报告实验验证候选SNP位点的功能;收集96例健康对照人群(对照组)和96例冠心病患者(疾病组)的外周血,利用Sanger法完成rs45448095位点测序,应用Logistic回归分析对照组人群中rs45448095不同基因型与血脂水平的关系,分析对照组和疾病组人群rs45448095不同基因型与冠心病风险的相关性。结果:中国汉族人群中PCSK9基因5′UTR区存在rs45448095(MAF>0.05),荧光报告基因实验显示在多个细胞系中(293T/L02)rs45448095-T比rs45448095-C表达显著下降(P<0.000 1);对照组人群中携带rs45448095-T基因多态性有更低水平的LDL水平[β(SE)=-0.353(0.158),t=-2.237,P=0.028],rs45448095不影响冠心病的发病风险。结论:PCSK9基因5′UTR区上一个有功能的rs45448095突变可影响血浆LDL水平。
Objective: To investigate the correlation of a PCSK9 5′-untranslated region(5′ UTR) rs45448095 polymorphism with plasma LDL-C levels and coronary heart disease(CHD). Methods: A process of bioinformatic analysis was used to analyze the PCSK9 5' UTR polymorphism in Chinese Han Group. Luciferase report assays were used to verify the function of variant. 96 controls and 96 people with Chinese Han CHD were collected. The logistic regression was used to analyze the correlation of a PCSK9 5′ UTR rs45448095 polymorphism with plasma LDL-C levels and CHD. Results: There was a common SNP rs45448095 in PCSK9 5' UTR. Both in L02 and 293 T cells, the luciferase reporter activity of the PGL3-T vector(containing T allele of rs45448095) decreased significantly(P<0.000 1) when compared with the PGL3-C construct(containing c allele of 45448095). Furthermore, the T allele was significantly related with the decreased LDL-C levels [β(SE)=-0.353(0.158),t=-2.237, P=0.028]. Finally, the case-control study suggested that the rs45448095 polymorphism is related with CHD risk. Conclusions: The potential functional variant in 5' UTR of PCSK9, rs45448095 can effect Chinese Han plasma LDL-C levels.

关键词(KeyWords)： 前蛋白转化酶枯草溶菌素9基因;5端非编码区;基因多态性;血浆低密度脂蛋白;冠心病
PCSK9,5'UTR;polymorphism;plasma LDL-C;coronary heart disease(CHD)

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金地区项目(31760294);; 贵州省科技厅项目[黔科合基础(2016)1120];贵州省科技厅项目[黔科合平台人才(2018)5608];; 贵阳市科技局项目[筑科合同(2017)5-14]

作者(Author): 张营,吴晟,鲍海龙,刘兴德
ZHANG Ying,WU Cheng,BAO Hailong,LIU Xingde

DOI: 10.19367/j.cnki.1000-2707.2020.01.008

参考文献(References)：

• [1] EMERIGN RISK FACTORS COLLABORATION.Major lipids,apolipoproteins,and risk of vascular disease[J].JAMA,2009,302(18):1993-2000.
• [2] CHOLESTEROL TREATMENT TRIALISTS COLLABORATION.Efficacy and safety of more intensive lowering of LDL cholesterol:a meta-analysis of data from 170 000 participants in 26 randomised trials[J].Lancet,2010,376(9753):1670-1681.
• [3] CANNON C P,BLAZING M A,GIUGLIANO R P,et al.Ezetimibe added to statin therapy after acute coronary syndromes[J].N Engl J Med,2015,372(25):2387-2397.
• [4] ALBERTI K G,ECKEL R H,GRUNDY S M,et al.Harmonizing the metabolic syndrome.A joint interim statement of the international diabetes federation task force on epidemiology and prevention;national heart,lung,and blood institute;American heart association;world heart federation;international atherosclerosis society;and international association for the study of obesity[J].Circulation,2009,120(16):1640-1645.
• [5] VENKETASUBRAMANIAN N,YOON B W,PANDIAN J,et al.Stroke epidemiology in South,East,and South-East Asia:a review[J],Journal of Stroke,2017,19(3):286-294.
• [6] ABIFADEL M,VARRET M,RABES J P,et al.Mutations in PCSK9 cause autosomal dominant hypercholesterolemia[J].Nat Genet,2003,34(2):154-156.
• [7] SABATINE M S,GIUGLIANO R P,WIVIOTT S D,et al.Efficacy and safety of evolocumab in reducing lipids and cardiovascular events[J].N Engl J Med,2015,372(16):1500-1509.
• [8] ROBINSON J G,FARNIER M,KREMPF M,et al.Efficacy and safety of alirocumab in reducing lipids and cardiovascular events[J].N Engl J Med,2015,372(16):1489-1499.
• [9] ZHANG Y,HU S L,LIU X D,et al.MiR-1322 regulates ChREBP expression by via binding a 3′-UTR variant (rs1051943)[J].J Cell Mol Med,2018,22(11):5322-5332.
• [10]COHEN J,PERTSEMLIDIS A,KOTOWSKI K,et al.Low LDL cholesterol in individuals of african descent resulting from frequent nonsense mutations in PCSK9[J].Nat Genet,2005,37(2):161-165.
• [11]CHEN S N,BALLANTYNE C M,GOTTO A M,et al.A common PCSK9 haplotype,encompassing the E670G coding single nucleotide polymorphism,is a novel genetic marker for plasma low-density lipoprotein cholesterol levels and severity of coronary atherosclerosis[J].J Am Coll Cardiol,2005,45(10):1611-1619.
• [12]COHEN J C,BOERWINKLE E,MOSLEY T H,et al.Sequence variations in PCSK9,low LDL,and protection against coronary heart disease[J].N Engl J Med,2006,354(12):1264-1272.
• [13]LAMBERT G,PETRIDES F,CHATELAIS M,et al.Elevated plasma PCSK9 level is equally detrimental for patients with nonfamilial hypercholesterolemia and heterozygous familial hypercholesterolemia,irrespective of low-density lipoprotein receptor defects[J].J Am Coll Cardiol,2014,63(22):2365-2373.
• [14]BERNELOT S J,NEELE A E,KROON J,et al.PCSK9 monoclonal antibodies reverse the pro-inflammatory profile of monocytes in familial hypercholesterolaemia[J].Eur Heart J,2017,38(20):1584-1593.
• [15]REYES G,PAVLYHA M,NGAI C,et al.effects of pcsk9 inhibition with alirocumab on lipoprotein metabolism in healthy humans[J].Circulation,2017,135(4):352-362.
• [16]HOPEWELL J C,MALIK R,VALDES E,et al.Differential effects of PCSK9 variants on risk of coronary disease and ischaemic stroke[J].Eur Heart J,2018,39(5):354-359.
• [17]BENN M,NORDESTGAARD B G,GRANDE P,et al.PCSK9 R46L,low-density lipoprotein cholesterol levels,and risk of ischemic heart disease:3 independent studies and meta-analyses[J].J Am Coll Cardiol,2010,55(25):2833-2842.
• [18]DING H,WU B,WANG H,et al.A novel loss-of-function DDAH1 promoter polymorphism is associated with increasedsusceptibility to thrombosis stroke and coronary heart disease[J].Circ Res,2010,106(6):1145-1152.
• [19]CUI G,LI Z,LI R,et al.A functional variant in APOA5/A4/C3/A1 gene cluster contributes to elevated triglycerides and severity of CAD by interfering with microRNA 3201 binding efficiency[J].J Am Coll Cardiol,2014,64(3):267-277.
• [20]KENT S T,ROSENSON R S,AVERY C L,et al.PCSK9 loss-of-function variants,low-density lipoprotein cholesterol,and risk of coronary heart disease and stroke:data from 9 studies of blacks and whites[J].Circ Cardiovasc Genet,2017,10(4):e001632.

文章评论(Comment)：