岳孟孟,于芳苹,赵迎春
YUE Mengmeng,YU Fangping,ZHAO Yingchun

• 1:南京医科大学上海松江临床医学院神经内科
• 2:南京医科大学上海松江临床医学院老年科

摘要(Abstract)：

目的:探讨凝血因子ⅡG20210A(FⅡG20210A)、ⅤG1691A(FⅤG1691A)及ⅦR353Q(FⅦ)基因多态性与上海汉族人群急性脑梗死(ACI)的相关性。方法:选取上海地区汉族ACI确诊患者180人作为ACI组[大动脉粥样硬化型(LAA,n=65)、小动脉硬化性或腔隙性(SAO,n=84)及心源性(CE,n=31)],同期健康体检的汉族人群150例作为对照组,收集2组研究对象的临床资料并比较;分别抽取2组研究对象入院或体检时清晨空腹外周血2 m L,采用聚合酶链式反应-限制性片段长度多态性(PCR-LDR)和多重连接酶检测(Multiple LDR)技术进行FⅡ、FⅤ、FⅦ基因分型检测并比较,采用活化蛋白C抵抗(APCR)试剂盒测定2组被检者外周血APCR阳性表达,采用Hardy-Weinberg平衡法检验样本的群体代表性,采用Logistic回归分析筛查ACI的危险因素。结果:ACI组与对照组研究对象的FⅡG20210A、FⅤG1691A及FⅦR353Q多态性符合Hardy-Weinberg平衡(P> 0. 05); FⅡG20210A的PCR产物经过HindⅢ酶切之后显示基因型为GG型(320 bp),Multi-LDR结果与PCR-RFLP结果一致,仅检测出GG基因型; FⅤG1691A的PCR产物经过MnlⅠ酶切显示基因型为GA型(353bp、205 bp及148 bp)和GG型(205 bp和148 bp); ACI组及对照组被检者均未检测出AA纯合突变基因型; FⅦR353Q的PCR产物经过MspⅠ酶切结果显示基因型为RQ型(291 bp、223 bp和68 bp)、RR型(223 bp和68 bp)和QQ型(291 bp),Multi-LDR结果与PCR-RFLP结果一致; ACI组FⅦR353Q基因型频率和等位基因频率与对照组比较,差别有统计学意义(P <0. 05); SAO组与对照组差异有统计学意义(P <0. 05),ACI组APCR阳性率高于对照组(P <0. 05);二分类Logistic回归分析结果显示,吸烟、高血压、低水平高密度脂蛋白、FⅦR353Q基因型及APCR是ACI的独立危险因素(P <0. 05)。结论:FⅡG20210A及FⅤG1691A基因多态性在上海汉族ACI患者中较少见,FⅦR353Q基因多态性与上海汉族人群ACI有关,其中Q等位基因可能是保护性因素。
Objective: To asses the association of coagulation factors Ⅱ G20210 A( FⅡ G20210 A),Ⅴ G1691 A( FⅤ G1691 A) and Ⅶ R353 Q( FⅦ) gene polymorphisms with acute cerebral infarction( ACI) among ethic Han in Shanghai. Methods: One hundred and eighty ethic Han patients diagnosed with ACI in Shanghai were selected as the ACI group,including aortic atherosclerosis( LAA,n = 65),arteriosclerosis or lacunarity( SAO,n = 84),and cardiogenicity( CE,n = 31),and 150 healthy ethic Han persons in the same period as control group. Clinical data of study subjects were collected and compared. 2 m L of fasting peripheral blood was collected in the early morning of the admission to our hospital or physical examination. Polymerase Chain reaction-restriction fragment length polymorphism( PCR-RFLP) and multiple ligase detection( Multiple LDR) assays were used to genotype FⅡ,FⅤ,FⅦ. Activated protein C resistance( APCR) kit was used to determine plasma APCR expression. The Hardy-Weinberg equilibrium was used to test the representativeness of the samples. Logistic regression analysis was used to screen risk factors for ACI. Results: The FⅡ G20210 A,FⅤ G1691 A and FⅦR353 Q gene polymorphisms were in accordance with Hardy-Weinberg equilibrium( P > 0. 05).Agarose gel electrophoresis analysis showed that the PCR product of FⅡ G20210 A digested with HindⅢ had only GG type( 320 bp),consistent with multi-LDR result. The PCR product of FⅤ G1691 A digested with Mnl I had GA type( 353 bp,205 bp and 148 bp) and GG type( 205 bp and 148 bp).AA type was not detected. The PCR product of F Ⅶ R353 Q digested with Msp Ⅰ had RQ type( 291 bp,223 bp and 68 bp),RR type( 223 bp and 68 bp) and QQ type( 291 bp),consistent with multi-LDR results. The genotype frequency and allele frequency of F Ⅶ R353 Q in ACI group were different from those in the control group( P < 0. 05),and the difference between the SAO group and the control group was statistically significant( P < 0. 05). The positive rate of APCR was higher in the ACI group than that in the control group( P < 0. 05). Binary logistic regression analysis showed that smoking,hypertension,low-level high-density lipoprotein,F Ⅶ R353 Q genotype and APCR were independent risk factors for ACI( P < 0. 05). Conclusion: F Ⅱ G20210 A and F Ⅴ G1691 A gene polymorphisms are rare in ACI patients among ethics Han in Shanghai. FⅦ R353 Q gene polymorphism is related to ACI among ethic Han in Shanghai,and Q allele may be a protective factor.

关键词(KeyWords)： 血液凝固因子;活化蛋白C抵抗;急性脑梗死;基因多态性;凝血因子ⅡG20210A;凝血因子Ⅴ G1691A;凝血因子ⅦR353Q
blood coagulation factors;activated protein C cresistance(APCR);acute cerebral infarction(ACI);gene polymorphism;coagulation factor Ⅱ G20210A(F Ⅱ G20210A);coagulation factor Ⅴ G1691A(FⅤ G1691A);coagulation factor Ⅶ R353Q(FⅦ R353Q)

Abstract:

Keywords:

基金项目(Foundation): 上海市松江区科学技术攻关项目(2017sjkjgg46);; 江苏省研究生科研与实践创新计划项目(KYCX18-1506)

作者(Author): 岳孟孟,于芳苹,赵迎春
YUE Mengmeng,YU Fangping,ZHAO Yingchun

DOI: 10.19367/j.cnki.2096-8388.2020.07.009

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