[1]胡小平,王志维,邓宏平,等.还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶-1基因重组真核表达质粒的构建及其对血管平滑肌细胞凋亡的影响[J].新乡医学院学报,2016,33(11):937-940.[doi:10.7683/xxyxyxb.2016.11.001]
　HU Xiao-ping,WANG Zhi-wei,DENG Hong-ping,et al.Construction of recombinant eukaryotic expression plasmid of nicotinamide adenine dinucleotide phosphate oxidase-1 gene and its effect on apoptosis of vascular smooth muscle cell[J].Journal of Xinxiang Medical University,2016,33(11):937-940.[doi:10.7683/xxyxyxb.2016.11.001]
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还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶-1基因重组真核表达质粒的构建及其对血管平滑肌细胞凋亡的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
33
期数:
2016年11
页码:
937-940
栏目:
基础研究
出版日期:
2016-11-05

文章信息/Info

Title:
Construction of recombinant eukaryotic expression plasmid of nicotinamide adenine dinucleotide phosphate oxidase-1 gene and its effect on apoptosis of vascular smooth muscle cell
作者:
胡小平; 王志维; 邓宏平; 吴红兵; 任宗力; 胡　锐
（武汉大学人民医院心血管外科，湖北　武汉　430060）
Author(s):
HU Xiao-ping; WANG Zhi-wei; DENG Hong-ping; WU Hong-bing; REN Zong-li; HU Rui
(Department of Cardiovascular Surgery,Renmin Hospital of Wuhan University,Wuhan 430060,Hubei Province,China)
关键词:
主动脉夹层; 氧化应激; 还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶-1; 质粒; 血管平滑肌细胞; 细胞凋亡
Keywords:
aortic dissection; oxidative stress; nicotinamide adenine dinucleotide phosphate oxidase-1; plasmid; vascular smooth muscle cell; apoptosis
分类号:
R543.1Q343.3+7
DOI:
10.7683/xxyxyxb.2016.11.001
文献标志码:
A
摘要:
目的　构建含人还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶-1（NOX1）基因重组真核表达质粒，探讨其对血管平滑肌细胞（VSMC）凋亡的影响。方法　通过美国国家生物技术信息中心（NCBI）数据库查找NOX1基因的mRNA序列，针对蛋白质编码区（CDS）设计引物，以人cDNA为模板扩增出NOX1基因全长CDS序列，经双酶切鉴定后，将NOX1基因与真核表达载体pcDNA3.1(+)连接，再经双酶切、测序鉴定。运用脂质体转染重组体pcDNA3.1(+)-NOX1至VSMC中，观察NOX1表达及VSMC凋亡情况。结果　重组体中NOX1基因片段和载体DNA大小与预期一致。经BLAST（the basic local alignment search tool）比对，与NCBI数据库中的NOX1基因序列具有高度的同源性。pcDNA3.1(+)-NOX1能促进VSMC中NOX1蛋白高表达，并促使VSMC的凋亡。结论　NOX1基因重组真核表达质粒能促使VSMC凋亡，为阐明NOX1在主动脉夹层中的作用及分子机制研究奠定了基础。
Abstract:
Objective　To construct the nicotinamide recombinant eukaryotic expression plasmid of nicotinamide adenine dinucleotide phosphate oxidase-1(NOX1) gene,and study its effect on apoptosis of vascular smooth muscle cell(VSMC).Methods　NOX1 gene was cloned using specific primers designed according to the mRNA sequence of human NOX1 gene from the National Center for Biotechnology Information (NCBI) database.The NOX1 gene was cloned to the pcDNA3.1(+) vector,and was identified using restriction digestion test and gene sequencing.The recombinant pcDNA3.1(+)-NOX1 was transfected into VSMC by lipofection,then the expression of NOX1 and apoptosis of VSMC in vitro were detected.Results　The obtained DNA fragments from recombinant pcDNA3.1(+)-NOX1 were in line with the NOX1 gene and pcDNA3.1(+),respectively.Sequence analysis using BLAST（the basic local alignment search tool） showed that its nucleotide sequence had high identity to the human NOX1 gene from NCBI database.pcDNA3.1(+)-NOX1 could induce over-expression of NOX1 in VSMC,and remarkably increase the apoptosis of VSMC.Conclusion　The recombinant eukaryotic expression plasmid of NOX1 gene can promote the apoptosis of VSMC,which lays the foundations for elucidating the role of NOX1 in aortic dissection.

参考文献/References:

［1］　孙立忠.主动脉外科学［M］.北京：人民卫生出版社，2012：127-150.
［2］　PACINI D,DI MARCO L,FORTUNA D,et al.Acute aortic dissection:epidemiology and outcomes［J］.Int J Cardiol,2013,167(6):2806-2812.
［3］　FU X J,PENG Y B,HU Y P,et al.NADPH oxidase 1 and its derived reactive oxygen species mediated tissue injury and repair［J］.Oxid Med Cell Longev,2014,2014:282854.
［4］　GAETAN G,CHRISTINE D,CANDICE T,et al.NOX1 deficiency protects from aortic dissection in response to angiotensin Ⅱ［J］.Hypertension,2007,50(1):189-196.
［5］　GAO F,GUO Z,SAKAMOTO M,et al.Fluid-structure interaction within a layered aortic arch model［J］.J Biol Phys,2006,32(5):435-454.
［6］　NAKASHIMA Y.Pathogenesis of aortic dissection:elastic fiber abnormalities and aortic medial weakness［J］.Ann Vasc Dis,2010,3(1):28-36.
［7］　LASSGUE B,GRIENDLING K K.NADPH oxidases:functions and pathologies in the vasculature［J］.Arterioscler Thromb Vasc Biol,2010,30(4):653-661.
［8］　GAVAZZI G,DEFFERT C,TROCME C,et al.NOX1 deficiency protects from aortic dissection in response to angiotensin Ⅱ［J］.Hypertension,2007,50(1):189-196.
［9］　ACAMPORA K B,NAGATOMI J,LANGAN E R,et al.Increased synthetic phenotype behavior of smooth muscle cells in response to in vitro balloon angioplasty injury model［J］.Ann Vasc Surg,2010,24(1):116-126.
［10］　ZHANG J,WANG L,FU W,et al.Smooth muscle cell phenotypic diversity between dissected and unaffected thoracic aortic media［J］.J Cardiovasc Surg (Torino),2013,54(4):511-521.
［11］　AILAWADI G,ELIASON J L,UPCHURCH G R J R.Current concepts in the pathogenesis of abdominal aortic aneurysm［J］.J Vasc Surg,2003,38(3):584-588.
［12］　DURDU S,DENIZ G C,BALCI D,et al.Apoptotic vascular smooth muscle cell depletion via bcl2 family of proteins in human ascending aortic aneurysm and dissection［J］.Cardiovasc Ther,2012,30(6):308-316.

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更新日期/Last Update: 2016-11-05