中国东北部汉族人群SLC22A3-LPAL2-LPA基因簇遗传变异及基因表达与冠心病关系的研究

发布时间：2018-04-17 05:03

本文选题：冠心病 + 脂蛋白(a)　；参考：《吉林大学》2015年博士论文

【摘要】：第一部分SLC22A3-LPAL2-LPA基因簇单核苷酸多态性与冠心病发病风险及血脂水平的关系目的：探索SLC22A3-LPAL2-LPA基因簇上的rs9346816、rs2221750、rs3127596、rs9364559、rs1367211、 rs6415085、rs9347438及rs9355296位点与中国东北部汉族人群冠心病发病风险及血脂水平的关系。方法：本研究采用病例对照研究方法。病例组和对照组分别为551例和544例。研究人群均为中国东北部汉族人群，且个体间无血缘关系。病例组人群筛选自2009年~2012年在吉林大学第一医院二部心血管内科确诊并进行住院治疗的冠心病患者。选择同期在吉林大学第一医院体检中心健康体检的正常人群为对照组。采用Sequenom MassArray系统对位于SLC22A3-LPAL2-LPA基因簇上的rs9346816、rs2221750、rs3127596、rs9364559、rs1367211、 rs6415085、rs9347438及rs9355296位点进行基因分型，同时收集研究对象的流行病学资料。采用SPSS16.0统计软件对一般资料、等位基因及基因型与冠心病关系、SNPs和冠心病危险因素的关联分析及不同基因型人群间血脂水平的关系；采用SHEsis软件分析单倍型与冠心病的关系；采用Haploview软件进行连锁不平衡分析。采用多元线性回归分析校正混杂因素对结果的影响。结果： 1.病例组中吸烟人数高于对照组，并且病例组中患有高血压病、糖尿病患者人数均多于对照组，病例组患者血清总胆固醇、低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、脂蛋白(a)水平高于对照组，并且以上差异均具有统计学意义（P0.05）。 2.校正混杂因素后，，SLC22A3-LPAL2-LPA基因簇上rs9364559位点基因型与等位基因在病例组和对照组人群中的分布仍存在显著性差异（X2=7.126, P=0.028；X2=4.436,P=0.039）。 3.单倍体型分析结果显示7个位点组成的单倍型中GAAAGTG和GGAAGTG与冠心病发病风险相关（P0.05），携带GAAAGTG单倍型人群冠心病的发病风险可能降低，但携带GGAAGTG单倍型人群冠心病的发病风险可能增高。由6个SNP位点构成的单倍型中有14种单倍型与冠心病的发病风险相关（P0.05），其中8种单倍型可能增加冠心病的发病风险，6种单倍型可能降低冠心病的发病风险。由5个SNP位点构成的单倍型中有39种单倍型与冠心病的发病风险相关（P0.05），其中21种单倍型可能增加冠心病的发病风险，18种单倍型可能降低冠心病的发病风险。由4个SNP位点构成的单倍型中有49种单倍型与冠心病的发病风险相关（P0.05），其中22种单倍型可能增加冠心病的发病风险，27种单倍型可能降低冠心病的发病风险。由3个SNP位点构成的单倍型中有39种单倍型与冠心病的发病风险相关（P0.05），其中19种单倍型可能增加冠心病的发病风险，20种单倍型可能降低冠心病的发病风险。由2个SNP位点构成的单倍型中有9种单倍型与冠心病的发病风险相关（P0.05），其中3种单倍型可能增加冠心病的发病风险，6种单倍型可能降低冠心病的发病风险。 4.位于SLC22A3基因上的rs9346816位点不同基因型间LDL-C、TC水平存在显著差异（P0.05），LPA基因上的rs6415085位点不同基因型之间Lp(a)、LDL-C水平存在显著差异（P0.05）。结论： 1．吸烟、高血压病、糖尿病、血清总胆固醇、低密度脂蛋白胆固醇、脂蛋白(a)水平可能与冠心病发病风险相关。 2. SLC22A3-LPAL2-LPA基因簇与中国东北部汉族人群冠心病发病风险及血脂水平相关。 3.在中国东北部汉族人群中位于LPA基因上的rs9364559位点可能参与冠心病的发生与发展。 4.多个SNPs位点同时存在可能对冠心病发病风险有不同的影响。第二部分中国东北部汉族人群SLC22A3-LPAL2-LPA基因簇mRNA表达水平与冠心病发病风险的关系目的：分析SLC22A3, LPAL2和LPA基因mRNA表达水平与冠心病发病风险的关系；阐述SLC22A3-LPAL2-LPA基因簇遗传变异对mRNA表达水平的影响。方法：选择2012年在吉林大学第一医院二部心血管内科确诊并进行住院治疗的92例冠心病患者为病例组，选择同期在吉林大学第一医院二部心血管内科进行诊治的非冠心病患者32例为对照组。采用实时荧光定量PCR方法对SLC22A3, LPAL2和LPA基因mRNA的表达水平进行测定。分析病例组与对照组之间及携带不同基因型病例组人群间目的基因mRNA表达水平的差异。以上统计学分析均采用SPSS16.0软件。结果： 1.病例组中吸烟、高血压病人数均高于对照组，病例组患者低密度脂蛋白胆固醇、高密度脂蛋白胆固醇、脂蛋白(a)水平高于对照组，并且以上差异均具有统计学意义（P0.05）。 2.与对照组人群相比，冠心病患者外周血中LPA和SLC22A3基因mRNA的表达水平显著增高（P0.05）；两组间外周血中LPAL2mRNA表达水平无显著差异（P0.05）。rs9346816、rs2221750、rs3127596、rs9364559、rs1367211、 rs6415085和rs9347438位点不同基因型患者外周血SLC22A3、LPAL2和LPAmRNA表达水平均无显著差异（P0.05）。结论： 1.外周血LPA和SLC22A3基因mRNA表达水平升高可能与中国东北部汉族人群冠心病的发病风险相关。 2.染色体6q26-27区域SLC22A3-LPAL2-LPA基因簇上rs9346816、rs2221750、rs3127596、rs9364559、rs1367211、rs6415085和rs9347438位点的多态性与冠心病患者外周血中SLC22A3、LPAL2、LPA基因mRNA的表达水平无关，因此，上述SNPs位点可能不是通过影响基因转录水平参与冠心病的发生。第三部分SLC22A3-LPAL2-LPA基因簇蛋白表达与冠心病发病风险的关系目的：研究SLC22A3-LPAL2-LPA基因簇蛋白表达与冠心病发病风险的关系；阐明SLC22A3-LPAL2-LPA基因簇遗传变异对其相邻基因蛋白表达水平的影响。方法：选择2012年在吉林大学第一医院二部心血管内科确诊并进行住院治疗的55例冠心病患者为病例组，选择同期在吉林大学第一医院二部心血管内科进行健康体检的正常人15例为对照组。采用western blot方法对外周血SLC22A3基因蛋白表达水平进行测定。用Quantity One软件对蛋白表达进行半定量分析。采用SPSS16.0统计软件包进行统计学分析。结果： 1.冠心病患者外周血中SLC22A3蛋白表达水平明显高于对照组，具有显著差异（P0.05）。 2.位于SLC22A3-LPAL2-LPA基因簇上rs9346816、rs2221750、rs3127596、rs9364559、rs1367211、rs6415085和rs9347438位点不同基因型间SLC22A3蛋白表达水平无显著性差异（P0.05）。结论： 1.冠心病患者外周血中SLC22A3蛋白表达水平增高可能是中国东北部汉族人群冠心病发病机制之一。 2. SLC22A3-LPAL2-LPA基因簇可能通过增加冠心病患者外周血中LPA和SLC22A3基因mRNA转录水平及SLC22A3蛋白表达水平影响冠心病发生。
[Abstract]:The relationship between the single nucleotide polymorphism of SLC22A3-LPAL2-LPA gene cluster and the risk of coronary heart disease and the level of blood lipids in the first part
Objective:
Objective to explore the relationship between rs9346816, rs2221750, rs3127596, rs9364559, rs1367211, rs6415085, rs9347438 and rs9355296 loci on the SLC22A3-LPAL2-LPA gene cluster and the risk of coronary heart disease and blood lipids in the Han population of Northeast China.
Method:
This study used a case-control study. The case group and the control group were 551 cases and 544 cases. The study population are Han nationality in Northeast Chinese population, and the individual unrelated cases. Population screening since 2009 ~2012 years in No.1 Hospital of Jilin University Department of cardiovascular medicine and diagnosis of two hospitalized patients with coronary heart disease in the same period. No.1 Hospital of Jilin University medical examination center of normal people as control group. Using Sequenom MassArray system on the SLC22A3-LPAL2-LPA gene cluster on rs9346816, rs2221750, rs3127596, rs9364559, rs1367211, rs6415085, rs9347438 and rs9355296 loci were genotyped and epidemiological data were collected at the same time. The research object of general data using statistical software SPSS16.0, and the allele the relationship between genotype and coronary heart disease, correlation analysis and different base SNPs and risk factors of coronary heart disease The relationship between blood lipid level was analyzed by SHEsis software. The relationship between haplotypes and coronary heart disease was analyzed by Haploview software. Linkage disequilibrium analysis was performed by using software. Multiple linear regression analysis was used to correct the influence of confounding factors on the results.
Result:
1. cases in the number of smokers was higher than the control group, and patients with hypertension, the number of patients with diabetes were more than the control group, serum total cholesterol group cases, low density lipoprotein cholesterol, high density lipoprotein cholesterol, lipoprotein (a) levels higher than the control group, and the above differences were statistically significant (P0.05).
2. after adjusting for confounding factors, there was significant difference in genotype distribution and allele distribution between rs9364559 locus and allele in SLC22A3-LPAL2-LPA group and control group (X2=7.126, P=0.028, X2=4.436, P=0.039).
3. haplotype analysis showed 7 loci haplotype in GAAAGTG and GGAAGTG and the risk for coronary heart disease (P0.05), relative risk of coronary heart disease with GAAAGTG haplotype groups may reduce the incidence of coronary heart disease, but the risk of carrying GGAAGTG haplotype population could increase. Consisting of 6 SNP loci haplotype in risk 14 haplotype and coronary heart disease (P0.05), of which 8 haplotypes may increase the risk of coronary heart disease, 6 haplotypes may reduce the risk of coronary heart disease. Consisting of 5 SNP loci haplotype in risk related 39 haplotypes with coronary heart disease (P0.05), of which 21 haplotypes may increase the risk of coronary heart disease, 18 haplotypes may reduce the risk of coronary heart disease. The risk of coronary heart disease and 49 haplotypes consisting of 4 SNP loci haplotype associated (P0.05) Among them, 22 haplotypes may increase the risk of coronary heart disease, 27 haplotypes may reduce the risk of coronary heart disease. Consisting of 3 SNP loci haplotype in risk related 39 haplotypes with coronary heart disease (P0.05), of which 19 haplotypes may increase the risk of coronary heart disease, 20 haplotypes may reduce the risk of coronary heart disease. Consisting of 2 SNP loci haplotype in risk related 9 haplotypes with coronary heart disease (P0.05), of which 3 haplotypes may increase the risk of coronary heart disease, 6 haplotypes may reduce the risk of coronary heart disease.
4., rs9346816 loci located on the SLC22A3 gene showed significant difference in LDL-C and TC levels among different genotypes (P0.05). Lp (a) and LDL-C level between different rs6415085 genotypes on LPA gene had significant difference (P0.05).
Conclusion:
1. smoking, hypertension, diabetes, serum total cholesterol, low density lipoprotein cholesterol (LDL), and lipoprotein (a) levels may be associated with the risk of coronary heart disease.
The 2. SLC22A3-LPAL2-LPA gene cluster is associated with the risk of coronary heart disease and the level of blood lipid in the Han population in the northeastern part of China.
3. the rs9364559 loci, located in the LPA gene in the Han population in northeastern China, may be involved in the development and development of coronary heart disease.
At the same time, more than 4. SNPs loci may have different effects on the risk of coronary heart disease.
The relationship between the expression level of SLC22A3-LPAL2-LPA gene cluster mRNA and the risk of coronary heart disease in the second part of the Han population in northeastern China
Objective:
Objective to analyze the relationship between mRNA expression level of SLC22A3, LPAL2 and LPA gene and the risk of coronary heart disease, and elaborate the influence of genetic variation of SLC22A3-LPAL2-LPA gene cluster on mRNA expression level.
Method:
In 2012 92 cases of hospitalized patients with coronary heart disease and the treatment of the case group in No.1 Hospital of Jilin University two Department of cardiovascular medicine were selected, were treated in No.1 Hospital of Jilin University Department of cardiovascular medicine two non coronary heart disease patients with 32 cases as control group. Using real-time quantitative PCR method of SLC22A3, the expression level of LPAL2 and LPA genes of mRNA were determined. The analysis between the case group and the control group and carrying different genotype differences in expression levels of mRNA gene groups. These cases statistical analysis was performed using SPSS16.0 software.
Result:
1., the number of smokers and hypertension patients in the case group was higher than that in the control group. The LDL cholesterol, high-density lipoprotein cholesterol and lipoprotein (a) level in the case group were higher than those in the control group, and the above differences were statistically significant (P0.05).
2. compared with the control group, the expression level of LPA and SLC22A3 in the peripheral blood of mRNA gene in patients with coronary heart disease were significantly higher (P0.05) between the two groups; the expression of LPAL2mRNA in peripheral blood level had no significant difference (P0.05).Rs9346816, rs2221750, rs3127596, rs9364559, rs1367211, SLC22A3, rs6415085 and rs9347438 loci in peripheral blood of patients with different genotypes LPAL2, and no significant difference in LPAmRNA expression level (P0.05).
Conclusion:
The increase of the expression level of LPA and SLC22A3 gene mRNA in peripheral blood may be associated with the risk of coronary heart disease in the Han population in northeastern China.
The 2. chromosome 6q26-27 region SLC22A3-LPAL2-LPA gene cluster on rs9346816, rs2221750, rs3127596, rs9364559, rs1367211, rs6415085 and rs9347438 loci polymorphism and coronary heart disease in patients with peripheral blood SLC22A3, LPAL2, LPA gene expression level of mRNA. Therefore, the SNPs locus may not be affected by the gene transcription level in coronary heart disease.
The relationship between the third part of SLC22A3-LPAL2-LPA gene cluster protein expression and the risk of coronary heart disease
Objective:
Objective to study the relationship between the expression of SLC22A3-LPAL2-LPA gene cluster and the risk of coronary heart disease (CHD), and clarify the influence of SLC22A3-LPAL2-LPA gene cluster genetic variation on the expression level of its adjacent gene protein.
Method:
In 2012 55 cases of hospitalized patients with coronary heart disease and the treatment of the case group in No.1 Hospital of Jilin University two Department of cardiovascular medicine diagnosis, normal people choose the same period the health examination in No.1 Hospital of Jilin University two Department of cardiovascular medicine. 15 cases as the control group. Using the Western blot method of peripheral blood SLC22A3 gene expression levels were determined by semi quantitative analysis of the protein. The expression of Quantity by One software. The data were analyzed by SPSS16.0 statistical software.
Result:
1. the expression level of SLC22A3 protein in peripheral blood of patients with coronary heart disease was significantly higher than that in the control group, with significant difference (P0.05).
2., on the SLC22A3-LPAL2-LPA gene cluster, there was no significant difference in the expression level of SLC22A3 protein between different genotypes among rs9346816, rs2221750, rs3127596, rs9364559, rs1367211, rs6415085 and rs9347438 locus (P0.05).
Conclusion:
1. the increase of SLC22A3 protein expression in peripheral blood of patients with coronary heart disease may be one of the pathogenesis of coronary heart disease in the Han population in northeastern China.
2., SLC22A3-LPAL2-LPA gene cluster may affect coronary heart disease by increasing the mRNA transcription level of LPA and SLC22A3 genes in peripheral blood and the expression level of SLC22A3 protein in patients with coronary heart disease.

【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2015
【分类号】：R541.4

【参考文献】