天津医药

天津医药 ›› 2019, Vol. 47 ›› Issue (8): 842-847.doi: 10.11958/20191585

• 临床研究 • 上一篇    下一篇

载脂蛋白E启动子区域-427T/C多态性与冠心病的关系及其对转录活性的影响

张蕊1，2，赵辉3，赵福梅4，任珉4，刘婷4，刘珊4，丛洪良2，4△   

  1. 1天津医科大学胸科临床学院（邮编300222）；2天津市胸科医院心内科；3天津市天津医院心内科；4天津市心血管病研究所
  • 收稿日期:2019-05-28 修回日期:2019-07-11 出版日期:2019-08-15 发布日期:2019-08-16
  • 通讯作者: 张蕊 E-mail:drzr2013@163.com
  • 作者简介:张蕊（1979），女，博士在读，主治医师，主要从事心血管疾病的相关研究
  • 基金资助:
    天津市科技计划项目

Relationship between apolipoprotein E promoter region-427T/C polymorphism and coronary heart disease and its effect on transcriptional activity

ZHANG Rui1,2, ZHAO Hui3, ZHAO Fu-mei4, REN Min4, LIU Ting4, LIU Shan4, CONG Hong-liang2, 4△   

  1. 1 Thoracic Clinical College, Tianjin Medical University, Tianjin 300222, China; 2 Department of Cardiology, Tianjin Chest Hospital; 3 Department of Cardiology, Tianjin Hospital; 4 Tianjin Institute of Cardiovascular Diseases
  • Received:2019-05-28 Revised:2019-07-11 Published:2019-08-15 Online:2019-08-16
  • Contact: Rui ZHANG E-mail:drzr2013@163.com

PDF (PC)

5617

摘要： 摘要：目的探讨载脂蛋白E（APOE）启动子区域-427T/C基因多态性与冠心病（CHD）发病的关系及其对APOE转录活性的影响。方法选取2016年10月—2017年9月在天津市胸科医院心内科住院的患者627例，分为CHD组415例，非CHD组212例。入院后检测总胆固醇（TC）、三酰甘油（TG）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）、脂蛋白a［Lp（a）］、载脂蛋白AI（APOAI）、载脂蛋白B（APOB）、尿酸（UA）水平；应用酶联免疫吸附法 （ELISA）测定血清APOE浓度；聚合酶链反应-限制性内切酶片段长度多态性（PCR-RFLP）技术检测-427T/C位点基因多态性，比较2组内TC+CC型和TT型患者血脂及载脂蛋白水平的差异；Logistic回归分析影响冠心病发病的危险因素。利用双荧光素酶报告基因表达载体pGL2构建携带APOE-427TT野生型重组质粒（TT组）、APOE/-427CC变异纯合型重组质粒（CC组），通过阳离子脂质体法将TT组、CC组、pGL2-Basic空载体质粒（对照组）分别和pRL-CMV内参质粒共转染至293T细胞内，检测3组细胞荧光素酶报告基因的荧光强度，以反映APOE的转录活性。结果（1）与非CHD组相比，CHD组的年龄、男性比例、吸烟比例、有糖尿病史、TG、Lp（a）、HCY、UA均出现升高，APOAI、HDL-C水平则降低，差异均有统计学意义（P＜0.01）。2组基因型分布符合Hardy-Weinberg遗传平衡（χ2=1.698，P＞0.05）， CHD组C等位基因频率较非CHD组增高（19.8% vs. 12.4%，χ2=8.959，P＜0.05）。（2）CHD组中TC+CC型患者APOB高 于TT组，APOA/B低于TT组（P＜0.05），非CHD组2种基因型间血脂及载脂蛋白水平差异无统计学意义（P＞0.05）。 多元Logistic回归分析结果表明，吸烟（OR=2.065，95%CI：1.369~3.115）、糖尿病（OR=3.355，95%CI：1.935~5.815）、UA升高（OR=1.008，95%CI：1.005~1.010）、携带APOE-427 C等位基因（OR=1.876，95%CI：1.185~2.969）是CHD发生的危险因素。（3）体外实验结果显示，CC组APOE转录活性明显低于TT组（P＜0.05）。结论APOE-427 C等位基因携带者是冠心病发病的危险因素，该位点通过影响APOE的转录活性及合成水平，进而影响血浆脂蛋白浓度及冠心病的发病。

关键词: 冠心病, 载脂蛋白E类, 多态性, 单核苷酸, 转录启动子, APOE-427T/C

Abstract: Abstract：Objective To investigate the relationship between apolipoprotein E (APOE) promoter region-427T/C gene polymorphism and the incidence of coronary heart disease (CHD) and its effect on the transcriptional activity of APOE.Methods A total of 627 patients admitted to the department of cardiology, Tianjin chest hospital from October 2016 toSeptember 2017 were selected and divided into CHD group (n=415) and non-CHD group (n=212). The total cholesterol (TC)，triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), lipoprotein a[Lp (a)], apolipoprotein AI (APOAI), apolipoprotein B (APOB) and uric acid (UA) levels were detected after admission. The serum concentration of APOE was determined by enzyme-linked immunosorbent assay (ELISA). Polymerase chain reactionrestriction enzyme fragment length polymorphism (PCR-RFLP) was used to detect gene polymorphism at the -427T/C site.Logistic regression analysis was used to evaluate risk factors of coronary heart disease. The dual luciferase report gene expression vector pGL2 was used to build carry APOE -427TT wild type recombinant plasmid (TT group) and APOE-427CC variation homozygous type recombinant plasmid (CC group). The TT group, CC group, pGL2-basic no-load plasmid (controlgroup) and pRL-CMV internal reference plasmid were co-transfected into 293T cells by cationic liposome method. The fluorescence intensity of luciferase reporter gene was detected in the three groups to reflect the transcriptional activity of APOE. Results (1) Compared with the non-CHD group, data of age, male ratio, smoking ratio, history of diabetes, TG,Lp(a), HCY and UA were significantly increased in the CHD group, while the levels of APOAI and HDL-C were significantly decreased (P＜0.01). The genotype distribution in the two groups was consistent with the Hardy-Weinberg genetic balance(χ2=1.698，P＞0.05), and the C allele frequency was increased in CHD group than that of non-CHD group (19.8% vs. 12.4%,χ2=8.959, P＜0.05). (2) The level of APOB of TC+CC patients was higher in the CHD group than that in the TT group (P＜0.05), and APOA / APOB was lower in the TT group (P＜0.05). There were no significant differences in lipid and apolipoprotein levels between the two genotypes in the non-CHD group (P＞0.05). Multivariate Logistic regression analysis showed that smoking (OR=2.065, 95%CI：1.369-3.115), diabetes (OR=3.355, 95%CI：1.935-5.815), higher level of UA (OR=1.008, 95%CI: 1.005-1.010), and carrying -427C allele (OR=1.876, 95% CI：1.185-2.969) were risk factors for CHD. (3)The results of in vitro experiments showed that the transcriptional activity of APOE was significantly lower in CC group than that in TT group (P＜0.05). Conclusion Carriers of APOE-427C gene are risk factors for the incidence of coronary heart disease, and this site affects the plasma lipoprotein concentration and the incidence of coronary heart disease by affecting the transcriptional activity and synthesis level of APOE.

Key words: coronary disease, apolipoproteins E, polymorphism, single nucleotide, transcription initiation site, APOE-427T/C