弹力蛋白原基因单核苷酸多态性与主动脉夹层的关系  

Association between single nucleotide polymorphisms of tropoelastin gene and aortic dissection

作　　者：戚悠飞[1] 舒畅[2] 刘飒华[1] 陈浩[1] 张文波[1] QI Youfei;SHU Chang;LIU Sahua;CHEN Hao;ZHANGWenbo(Department ofVascularSurgery,Hainan GeneralHospital,Haikou 570311;Department ofVascularSurgery,Second XiangyaHospital,Central SouthUniversity,Changsha 410011,China)

机构地区：[1]海南省人民医院血管外科,海口570311 [2]中南大学湘雅二医院血管外科,长沙410011

出　　处：《中南大学学报（医学版）》2021年第5期458-466,共9页Journal of Central South University ：Medical Science

基　　金：海南省自然科学基金(813205);海南省社会发展科技专项(2015SF04)。

摘　　要：目的:通过检测弹力蛋白原(tropoelastin)基因单核苷酸多态性(single nucleotide polymorphisms,SNPs)位点表达、tropoelastin mRNA和弹性蛋白与弹力纤维水平,探讨tropoelastin基因SNPs与主动脉夹层(aortic dissection,AD)的关系。方法:留取96例AD患者(AD组)和95例对照组患者的外周血和主动脉组织标本,提取外周血DNA,应用MassARRAY技术进行tropoelastin基因SNPs检测,并构建单体型;采用real-time PCR和蛋白质印迹法分别检测主动脉中膜层tropoelastin mRNA和弹性蛋白的表达;采用EVG(Elastin Van Gieson)染色观察主动脉管形态和壁弹力纤维的分布。比较tropoelastin基因各SNPs位点基因型和单体型在两组间频率的分布,以及tropoelastin m RNA、弹性蛋白和弹力纤维在两组中表达的差异。结果:两组共检测tropoelastin基因SNPs位点7个,5个位点呈多态性,其中rs2071307(G/A)、rs34945509(C/T)和rs17855988(G/C)位点频率分布在两组之间差异均有统计学意义(均P<0.05)。3个SNP位点[rs2071307(G/A)、rs34945509(C/T)和rs17855988(G/C)]的单体型频率分布,在两组之间差异均有统计学意义(均P<0.01)。Real-time PCR和蛋白质印迹法结果显示AD组主动脉中膜层tropoelastin m RNA和弹性蛋白的表达量均显著低于对照组(均P<0.05)。EVG染色可见AD组主动脉中膜层撕裂,弹力纤维破碎、断裂,且排列不规则;而对照组主动脉中膜层结构完整、层次分明,弹力纤维排列紧密、有序。结论:Tropoelastin基因rs2071307(G/A)、rs34945509(C/T)、rs17855988(G/C)位点的多态性改变可能最终会影响弹力纤维的合成,并有可能在AD的发生中起重要作用。Objective:To evaluate the relation between single nucleotide polymorphisms(SNPs)of tropoelastin gene and aortic dissection(AD)via identifying SNPs in the tropoelastin gene,and to detect the level of tropoelastin mRNA,elastin and elastic fibers.Methods:The specimens of the AD group(n=96)and the control group(n=95),including their blood and aortic wall tissues,were collected.DNA was extracted from the blood samples in the 2 groups,and the SNPs in the tropoelastin gene were examined by the MassARRAY genotyping technique,and their haplotypes were constructed by PHASE software.The expression of tropoelastin mRNA and elastin in the aortic tunica media was respectively detected by real-time PCR or Western blotting.Elastin Van Gieson(EVG)staining was used to observe the shape of aortic tunica media and clarify the distribution of elastic fibers.The frequency of genotypes and haplotypes of SNP loci in the tropoelastin gene was analyzed and compared between the 2 groups,and the expression of tropoelastin mRNA,elastin and elastic fibers were also compared.Results:Seven SNP loci of the tropoelastin gene were detected in these samples.Among them,5 SNP loci were polymorphic.The frequency of 3 SNP loci[rs2071307(G/A),rs34945509(C/T)and rs17855988(G/C)]was significantly different between the AD group and the control group(all P<0.05).There were significantly different in the haplotypes frequency of rs2071307(G/A),rs34945509(C/T)and rs17855988(G/C)between the 2 groups(all P<0.01).Real-time PCR and Western blotting showed that the relative expression of tropoelastin mRNA and elastin in the aortic tunica media in the AD group was significantly lower than that in the control group(P<0.05).EVG staining showed that the aortic tunica media was torn,the morphology and structure of elastic fibers were broken,cracked,and disordered in the AD group,while the aortic tunica media was in complete structure and well arrangement.The elastic fibers were presented closely and orderly in the control group.Conclusion:The polymorphisms of rs2071307(

关 键 词：弹力蛋白原基因 主动脉夹层 单核苷酸多态性 单体型 弹性蛋白 

分 类 号：R543.1[医药卫生—心血管疾病]