中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2009年
36期
2574-2577
,共4页
李连喜%陶征%董雪红%梁文昌%杨志红%牟波%包玉倩%王琛%贾伟平%胡仁明
李連喜%陶徵%董雪紅%樑文昌%楊誌紅%牟波%包玉倩%王琛%賈偉平%鬍仁明
리련희%도정%동설홍%량문창%양지홍%모파%포옥천%왕침%가위평%호인명
糖尿病血管病%分子克隆%基因%计算生物学
糖尿病血管病%分子剋隆%基因%計算生物學
당뇨병혈관병%분자극륭%기인%계산생물학
Diabetic angiopathies%Molecular cloning%Gene%Computational biology
目的 克隆与糖尿病大血管病变相关的新基因C5orf21基因,初步研究C50rf21基因在糖尿病大血管病变中的作用.方法 应用RT-PCR从人主动脉组织中克隆出C5orf21基因并经酶切、测序鉴定;研究C50rf21基因的组织表达谱、血管细胞表达谱以及高糖高脂对巨噬细胞中C5orf21基因表达的影响,并对C5orf21基因及蛋白功能进行分析.结果 成功克隆出CSorf21基因,人主动脉组织中存在5种C5orf21基因剪切体,编码框长度分别为1251、1113、894、810、810 bp,第3、5种剪切体GenBank数据库中尚未有收录,其中第4、第5种剪切体的蛋白编码框相同,均为810 bp,其差别主要在5′非翻译区碱基的不同,生物信息学分析发现C5orf21基因定位于染色体5q15,C5orf21含有Arb2结构域,可能与H3组蛋白第9位赖氨酸的甲基化有关;C5orf21组织表达较为广泛,以脂肪和主动脉组织表达最高;人主动脉内皮细胞、平滑肌细胞及巨噬细胞均有C5orf21基因的表达;高葡萄糖、油酸及LDL均上调巨噬细胞中C5orf21基因的表达.结论 首次在mRNA水平证实了C5orf21基因的存在,C5orf21基因表达的改变与糖尿病大血管病变有关.
目的 剋隆與糖尿病大血管病變相關的新基因C5orf21基因,初步研究C50rf21基因在糖尿病大血管病變中的作用.方法 應用RT-PCR從人主動脈組織中剋隆齣C5orf21基因併經酶切、測序鑒定;研究C50rf21基因的組織錶達譜、血管細胞錶達譜以及高糖高脂對巨噬細胞中C5orf21基因錶達的影響,併對C5orf21基因及蛋白功能進行分析.結果 成功剋隆齣CSorf21基因,人主動脈組織中存在5種C5orf21基因剪切體,編碼框長度分彆為1251、1113、894、810、810 bp,第3、5種剪切體GenBank數據庫中尚未有收錄,其中第4、第5種剪切體的蛋白編碼框相同,均為810 bp,其差彆主要在5′非翻譯區堿基的不同,生物信息學分析髮現C5orf21基因定位于染色體5q15,C5orf21含有Arb2結構域,可能與H3組蛋白第9位賴氨痠的甲基化有關;C5orf21組織錶達較為廣汎,以脂肪和主動脈組織錶達最高;人主動脈內皮細胞、平滑肌細胞及巨噬細胞均有C5orf21基因的錶達;高葡萄糖、油痠及LDL均上調巨噬細胞中C5orf21基因的錶達.結論 首次在mRNA水平證實瞭C5orf21基因的存在,C5orf21基因錶達的改變與糖尿病大血管病變有關.
목적 극륭여당뇨병대혈관병변상관적신기인C5orf21기인,초보연구C50rf21기인재당뇨병대혈관병변중적작용.방법 응용RT-PCR종인주동맥조직중극륭출C5orf21기인병경매절、측서감정;연구C50rf21기인적조직표체보、혈관세포표체보이급고당고지대거서세포중C5orf21기인표체적영향,병대C5orf21기인급단백공능진행분석.결과 성공극륭출CSorf21기인,인주동맥조직중존재5충C5orf21기인전절체,편마광장도분별위1251、1113、894、810、810 bp,제3、5충전절체GenBank수거고중상미유수록,기중제4、제5충전절체적단백편마광상동,균위810 bp,기차별주요재5′비번역구감기적불동,생물신식학분석발현C5orf21기인정위우염색체5q15,C5orf21함유Arb2결구역,가능여H3조단백제9위뢰안산적갑기화유관;C5orf21조직표체교위엄범,이지방화주동맥조직표체최고;인주동맥내피세포、평활기세포급거서세포균유C5orf21기인적표체;고포도당、유산급LDL균상조거서세포중C5orf21기인적표체.결론 수차재mRNA수평증실료C5orf21기인적존재,C5orf21기인표체적개변여당뇨병대혈관병변유관.
Objective To clone a novel diabetic angiopathy related protein gene- C5orf21 and study its roles in diabetic macroangiopathy. Methods The open reading frame (ORF) of C5orf21 gene was cloned into vector from human aortic tissues by a RT-PCR-based approach and identified by enzyme-cutting and sequencing. The structure and function of C5orf21 gene and protein were further analyzed by bioinformatics technology. The mRNA expression of C5orf21 gene in human tissues and in vascular cells was analyzed by RT-PCR. RT-PCR was used to observe the effect of high glucose, low-density lipoprotein (LDL) and free fatty acid (FFA) upon the expression of C5orf21 gene in macrophnges. Results C5orf21 gene was successfully inserted into plh-ive vector and identified for the first time at the level of mRNA. There are five C5orf21 gene splice variants in human aortic tissue and their length of ORF are 1251, 1113, 894, 810 and 810 bp respectively. Two kinds of splice variants have yet to be included in GenBank database. Two kinds of splice variants have the same ORF and their differences are mainly in the bases in the 5' untranslated region. Bioinformatics analysis found that C5orf21 gene was located in chromosome 5q15 and C5orf21 protein contained Arb2 domain associated with histone H3 lysine 9 methylation. C5orf21 gene was normally expressed in many tissues. Fat and aortic tissues had the highest expression. The expression of C5orf21 gene could be detected in human aortic endothelial cell, aortic smooth muscle cell and macrophages. High glucose, LDL and FFA (esp. high glucose) up-regulated the expression of C5orf21 gene in macrophage. Conclusion C5orf21 gene contains five splice variants and it is identified for the first time at the level of mRNA. The changes of C5orf21 gene expression are correlated with diabetic macroangiopathy.
