中华心血管病杂志
中華心血管病雜誌
중화심혈관병잡지
Chinese Journal of Cardiology
2014年
4期
284-289
,共6页
郑璐%张俊%牟琴峰%张晓膺%罗光华
鄭璐%張俊%牟琴峰%張曉膺%囉光華
정로%장준%모금봉%장효응%라광화
载脂蛋白类%冠状动脉疾病%多态性,单核苷酸%启动区(遗传学)
載脂蛋白類%冠狀動脈疾病%多態性,單覈苷痠%啟動區(遺傳學)
재지단백류%관상동맥질병%다태성,단핵감산%계동구(유전학)
Apolipoproteins%Coronary disease%Polymorphism,single nucleotide%Promoter regions (genetics)
目的 检测载脂蛋白M(apoM)基因启动子区域的单核苷酸多态性(SNP),探讨多态性位点对转录活性的影响以及与冠心病易感性的关系.方法 将受试者分为2组:冠心病组,经冠状动脉造影证实1或1支以上血管直径狭窄50％以上者206例,男165例,女41例,平均年龄(61.9±9.2)岁;对照组,冠状动脉造影无病变者209例,男157例,女性52例,平均年龄(60.4±9.1)岁.设计2对引物用以扩增apoM启动子区序列,采用PCR产物直接测序法进行序列分析.比较冠心病组和正常组间各基因型及等位基因频率分布的差异.进一步利用基因重组和定点突变技术体外观察SNP对apoM基因启动子转录活性的影响.结果 在apoM启动子区-724位发现1个新的胞嘧啶核苷酸缺失突变(-724delC).冠心病组-724del等位基因频率高于对照组(8.0％比4.1％,OR=2.054,95％ CI 1.125～3.749,P=0.017),-724del等位基因携带者血浆apoM的表达水平降低,总胆固醇水平明显高于C/C基因型[(6.04±0.90) mmol/L比(4.95±1.00) mmol/L,P＜0.01].与apoM野生型启动子比较,-724C位点发生缺失突变后,apoM启动子活性明显下降(相对荧光素酶活性比值:1.13±0.25比2.11±0.15,P=0.009).结论 新发现的-724delC缺失突变可降低apoM启动子活性,下调apoM蛋白表达水平,增加冠心病的发病风险.
目的 檢測載脂蛋白M(apoM)基因啟動子區域的單覈苷痠多態性(SNP),探討多態性位點對轉錄活性的影響以及與冠心病易感性的關繫.方法 將受試者分為2組:冠心病組,經冠狀動脈造影證實1或1支以上血管直徑狹窄50％以上者206例,男165例,女41例,平均年齡(61.9±9.2)歲;對照組,冠狀動脈造影無病變者209例,男157例,女性52例,平均年齡(60.4±9.1)歲.設計2對引物用以擴增apoM啟動子區序列,採用PCR產物直接測序法進行序列分析.比較冠心病組和正常組間各基因型及等位基因頻率分佈的差異.進一步利用基因重組和定點突變技術體外觀察SNP對apoM基因啟動子轉錄活性的影響.結果 在apoM啟動子區-724位髮現1箇新的胞嘧啶覈苷痠缺失突變(-724delC).冠心病組-724del等位基因頻率高于對照組(8.0％比4.1％,OR=2.054,95％ CI 1.125～3.749,P=0.017),-724del等位基因攜帶者血漿apoM的錶達水平降低,總膽固醇水平明顯高于C/C基因型[(6.04±0.90) mmol/L比(4.95±1.00) mmol/L,P＜0.01].與apoM野生型啟動子比較,-724C位點髮生缺失突變後,apoM啟動子活性明顯下降(相對熒光素酶活性比值:1.13±0.25比2.11±0.15,P=0.009).結論 新髮現的-724delC缺失突變可降低apoM啟動子活性,下調apoM蛋白錶達水平,增加冠心病的髮病風險.
목적 검측재지단백M(apoM)기인계동자구역적단핵감산다태성(SNP),탐토다태성위점대전록활성적영향이급여관심병역감성적관계.방법 장수시자분위2조:관심병조,경관상동맥조영증실1혹1지이상혈관직경협착50％이상자206례,남165례,녀41례,평균년령(61.9±9.2)세;대조조,관상동맥조영무병변자209례,남157례,녀성52례,평균년령(60.4±9.1)세.설계2대인물용이확증apoM계동자구서렬,채용PCR산물직접측서법진행서렬분석.비교관심병조화정상조간각기인형급등위기인빈솔분포적차이.진일보이용기인중조화정점돌변기술체외관찰SNP대apoM기인계동자전록활성적영향.결과 재apoM계동자구-724위발현1개신적포밀정핵감산결실돌변(-724delC).관심병조-724del등위기인빈솔고우대조조(8.0％비4.1％,OR=2.054,95％ CI 1.125～3.749,P=0.017),-724del등위기인휴대자혈장apoM적표체수평강저,총담고순수평명현고우C/C기인형[(6.04±0.90) mmol/L비(4.95±1.00) mmol/L,P＜0.01].여apoM야생형계동자비교,-724C위점발생결실돌변후,apoM계동자활성명현하강(상대형광소매활성비치:1.13±0.25비2.11±0.15,P=0.009).결론 신발현적-724delC결실돌변가강저apoM계동자활성,하조apoM단백표체수평,증가관심병적발병풍험.
Objective To investigate the association between genetic polymorphisms of proximal promoter region of apolipoprotein M (apoM) gene and susceptibility of coronary artery diseases (CAD) in Han Chinese population.Methods Two pairs of primers were designed according to the sequence (GenBank accession nos.EU030444.1) and the PCR products of apoM proximal promoter region were directly sequenced.Two hundred and six patients [165 males,mean age (61.9 ± 9.2) years old] diagnosed with CAD according to the results of angiography (a lesion was classed as being significant when stenosis was more than 50％) were enrolled in the present study,209 age-and gender-matched patients[157 males,mean age (60.4 ± 9.1) years old] without CAD according to the results of angiography were selected as the control group.The allelic frequencies and genotype distributions of polymorphism in CAD and nonCAD patients were analyzed.Furthermore the wide-type and mutant promoter region of apoM were cloned into the luciferase expression vector pGL3,respectively.Luciferase reporter assay was used to detect the activity of apoM promoter.Results A new deletion mutation-724delC in apoM promoter was found.The frequency of Del C allele was 8.0％ in CAD patients and only 4.1％ in the non-CAD controls (OR =2.054,95％CI 1.125-3.749,P =0.017).The mean TC level was lower in groups with wide-type homozygotes compared to the mutant allele carriers [(6.04 ± 0.90) mmol/L vs.(4.95 ± 1.00) mmol/L,P ＜ 0.01].-724delC mutant showed obvious decreased luciferase activities (1.13 ±0.25 vs.2.11 ±0.15,P =0.009).Conclusion It is reasonable to speculate that-724delC could affect the activity of the apoM promoter and downregulate apoM expressions,therefore,influence the susceptibility of CAD in this patient cohort.
