本文選題：冠心病 + 單核苷酸多態(tài)性　； 參考：《北京協(xié)和醫(yī)學(xué)院》2015年博士論文

【摘要】：背景：冠心病是危害人類健康影響壽命的重要公共衛(wèi)生問題之一。已有大量分子流行病學(xué)和遺傳學(xué)研究顯示,冠心病是一種復(fù)雜的多基因疾病,多種遺傳因素和環(huán)境因素與冠心病有關(guān),尤其是與脂質(zhì)代謝相關(guān)的多個基因在冠心病發(fā)生發(fā)展中起重要作用。其中,從2008年至2013年,全基因組關(guān)聯(lián)性研究(genome-wide association studies, GWAS)報道了近200個與脂代謝相關(guān)位點,得到了豐富的研究成果。本研究根據(jù)GWAS在脂代謝和冠心病中的研究結(jié)果,同時結(jié)合本實驗室的前期研究工作,共篩選了脂蛋白a (Lipoprotein(a), LPA)基因、凝集素(clusterin, CLU)基因、磷脂轉(zhuǎn)運蛋白(phospholipid transfer protein, PLTP)基因和乳鐵蛋白(lactoferrin, LTF)基因這四個候選基因進(jìn)行其基因單核苷酸多態(tài)性研究。目的：探討LPA、CLU、PLTP、LTF基因單核苷酸多態(tài)性與冠心病的相關(guān)性及相關(guān)代謝指標(biāo)的關(guān)系。方法：1、研究對象(1)非冠心病組：選擇367名冠狀動脈造影所示所有冠狀動脈主要分支直徑狹窄均不大于25%的人群入選至非冠心病組。(2)冠心病組：選擇496名冠狀動脈造影所示至少一支心外膜下冠狀動脈主要分支直徑狹窄大于75%的人群入選至冠心病組。2、候選基因和SNPs位點的選擇：本文根據(jù)本實驗室前期工作的研究成果,同時結(jié)合GWAS在冠心病中的研究結(jié)果,確定LPA、CLU、PLTP和LTF這四個候選基因進(jìn)行單核苷酸多態(tài)性研究。SNPs位點選擇上,首先選取基因功能區(qū)可能對蛋白質(zhì)水平產(chǎn)生影響的所有SNPs位點,再綜合該位點在中國北方漢族人群中最小等位基因頻率(Minor Allele Frequency, MAF)和文獻(xiàn)檢索對該位點其他人群驗證結(jié)果報道等進(jìn)行篩選,最終篩選出了10個有意義的SNPs位點。3、SNPs位點的檢測方法：使用基質(zhì)輔助激光解吸附電離飛行時間質(zhì)譜分析質(zhì)譜技術(shù)(matrix-assisted laser desorption/ionization-time of flight, MALDI-TOF)進(jìn)行分型檢測。4、統(tǒng)計分析方法：應(yīng)用SPSS 20.0軟件對計數(shù)資料進(jìn)行卡方檢驗,對正態(tài)分布的計量資料進(jìn)行單因素方差分析,對非正態(tài)分布的計量資料進(jìn)行秩和檢驗,當(dāng)P值小于0.05認(rèn)為具有顯著的統(tǒng)計學(xué)差異。結(jié)果：1、Hardy-Winberg平衡檢驗：9個SNPs位點符合H-W平衡。2、冠心病組與非冠心病組間關(guān)聯(lián)分析結(jié)果：(1)組間等位基因頻率的關(guān)聯(lián)分析結(jié)果：位于LPA基因上的rs3798220位點(C/T)在等位基因頻率方面具有顯著統(tǒng)計學(xué)差異(P=0.049),余位點未見顯著性差異。(2)組間基因型頻率的關(guān)聯(lián)分析結(jié)果：9個SNPs位點均未見顯著差異(P0.05)。(3)組間基因模型中基因型頻率的關(guān)聯(lián)分析結(jié)果：顯性模型中LPA基因的rs3798220位點(CC+CT型vs TT型)具有顯著的統(tǒng)計學(xué)差異(p=0.043),余位點未見顯著差異。隱性模型中CLU基因上的rs3216167位點(DEL型vs A. DEL+AA型)在兩組受試者間具有顯著的統(tǒng)計學(xué)差異P=0.047,余位點未見顯著性差異(P0.05)。(4)單倍體分析：rs3798220和rs1801693位點、rs3216167、rs7982、rs1532278位點之間以及rs378114和rs394643位點之間為連鎖不平衡區(qū)域,各連鎖不平衡區(qū)域均與冠心病不相關(guān)。3、不同基因型間計量指標(biāo)的比較：(1)LPA基因：rs3798220位點中血清LPA具有顯著的統(tǒng)計學(xué)差異(P=0.001);rs1801693位點中未見有顯著性差異的血清代謝指標(biāo)；rs7765803位點中血清LPA具有顯著的統(tǒng)計學(xué)差異[P=0]；(2)CLU基因：rs3216167位點中血清LPA和HbAlc具有顯著的統(tǒng)計學(xué)差異(P值分別為0.032,0.029)；rs7982位點中血清TC、LDL具有顯著的統(tǒng)計學(xué)差異(P值分別為0.069,0.051);rsl532278位點中血清TC水平、LDL、APOB具有顯著的統(tǒng)計學(xué)差異(P值分別為0.048,0.039和0.052)；(3)LTF基因：rs1126477位點血清TG具有顯著的統(tǒng)計學(xué)差異(P=0.011)；(4) PLTP基因：rs378114位點中未見有顯著差異的血清代謝指標(biāo)；rs394643位點中血清APOB具有顯著的統(tǒng)計學(xué)差異(P=0.011)4、SNP位點等位基因變化引起冠心病風(fēng)險變化程度rs3798220位點中,CT+CC型發(fā)生冠心病的風(fēng)險比TT增加了1.5倍。該位點能夠解釋0.7%的冠心病病因(Nagelkerke R2=0.07)。5、血LPA和冠心病關(guān)系的關(guān)聯(lián)指標(biāo)根據(jù)rs3798220基因型分組,比較基因型-疾病,基因型-血清LPA水平,推導(dǎo)出血清LPA水平和冠心病關(guān)系的關(guān)聯(lián)指標(biāo)OELPA-CAD=1.503K 14.64,即血清LPA變化K個單位時發(fā)生冠心病的效應(yīng)值。結(jié)論：1、LPA基因：rs3798220位點可能通過影響血清LPA水平影響冠心病發(fā)生；rs7765803位點可能影響血LPA水平；2、CLU基因：rs3216167位點可能影響血清LPA水平,并與冠心病發(fā)生相關(guān)；rs7982位點可能影響血TC、LDL水平；rs1532278位點可能影響血TC、LDL、APOB水平；3、LTF基因：rs1126477位點可能影響血TG水平4、PLTP基因：rs394643位點可能影響血APOB水平5、血清LPA變化K個單位史冠心病風(fēng)險ORLPA-CAD=1.503K/14.64
[Abstract]:Background: coronary heart disease is one of the important public health problems which harm the life of human health. A large number of molecular epidemiology and genetic studies have shown that coronary heart disease is a complex polygenic disease. Many genetic and environmental factors are associated with coronary heart disease, especially the multiple genes associated with lipid metabolism in coronary heart disease. Development plays an important role. From 2008 to 2013, the genome-wide association studies (GWAS) reported nearly 200 lipid metabolism related sites and obtained abundant research results. This study, based on the results of GWAS in lipid metabolism and coronary heart disease, combined with previous research work in the laboratory. A total of four candidate genes, such as lipoprotein a (Lipoprotein (a), LPA) gene, clusterin (CLU) gene, phospholipid transporter (phospholipid transfer protein, PLTP) gene and lactoferrin (lactoferrin, LTF) gene, were screened to study the single nucleotide polymorphisms of the genes. Association of acid polymorphism with coronary heart disease and related metabolic indicators. Methods: 1, subjects (1) non coronary heart disease group: selected 367 coronary arteriography of all the main branches of coronary artery stenosis not more than 25% of the population were selected to non coronary heart disease group. (2) coronary heart disease group: select 496 coronary arteriography at least to show at least The selection of.2, candidate genes and SNPs loci in a group of coronary artery disease with a diameter of more than 75% of the main branch of the epicardium coronary artery was selected. According to the results of earlier work in this laboratory, and combined with the results of GWAS in coronary heart disease, the four candidate genes of LPA, CLU, PLTP and LTF were identified for single nucleotide polymorphisms. In the selection of.SNPs loci, first select all SNPs loci which may affect the protein level of the gene function region, and then select the minimum allele frequency (Minor Allele Frequency, MAF) in the Han population in northern China and the literature retrieval to screen the results of the other populations of the loci. 10 meaningful SNPs loci,.3, SNPs loci were selected for detection of SNPs loci: using matrix assisted laser desorption ionization time of flight mass spectrometry analysis mass spectrometry (matrix-assisted laser desorption/ionization-time of flight, MALDI-TOF) for typing detection.4, statistical analysis method: using SPSS 20 software to carry out counting data Chi square test, a single factor variance analysis was carried out on the measurement data of the normal distribution. The rank sum test of the non normal distribution data was carried out. When the P value was less than 0.05, there were significant statistical differences. Results: 1, Hardy-Winberg balance test: 9 SNPs loci were conformed to H-W balance.2, the correlation analysis between coronary heart disease group and non coronary heart disease group was found. (1) correlation analysis of allele frequencies between groups: the rs3798220 loci (C/T) located on the LPA gene had significant statistical differences (P=0.049), and there was no significant difference in the residual loci. (2) the correlation analysis of genotype frequencies between groups had no significant difference between 9 SNPs loci (P0.05). (3) gene modes among groups. The correlation analysis results of genotype frequency in the model: the rs3798220 locus (CC+CT type vs TT type) of the LPA gene in the dominant model had significant statistical differences (p=0.043), and there was no significant difference in the residual loci. The rs3216167 loci on the CLU gene (DEL vs A. DEL+AA type) in the recessive model had significant statistical differences between the two groups. There was no significant difference in the residual loci (P0.05). (4) haploid analysis: rs3798220 and rs1801693 loci, rs3216167, rs7982, rs1532278 sites and rs378114 and rs394643 loci were interlinked unbalance regions, and the interlinked unbalance regions were not associated with coronary heart disease, and the comparison between different genotypes was compared: (1) LPA gene: RS3 There were significant statistical differences in serum LPA in the 798220 loci (P=0.001); there was no significant difference in serum metabolism index in the rs1801693 locus, and there was a significant statistical difference between the serum LPA and [P=0] in the rs7765803 locus. (2) the serum LPA and HbAlc of the rs3216167 site were statistically significant (P value was 0.032,0.02). 9): the serum TC and LDL in the rs7982 site had significant statistical differences (P values were 0.069,0.051), and the serum TC level, LDL, and APOB in the rsl532278 locus had significant statistical differences (P values were 0.048,0.039 and 0.052), and (3) the LTF genes were statistically significant; (4) There was no significant difference in serum metabolic index in the 114 loci. The serum APOB in the rs394643 site had significant statistical difference (P=0.011) 4. The change of SNP allele caused the risk of coronary heart disease in rs3798220 site, and the risk of coronary heart disease of type CT+CC was 1.5 times higher than that of TT. This site could explain 0.7% of coronary heart disease. The correlation index of the relationship between (Nagelkerke R2=0.07).5, blood LPA and coronary heart disease is based on the grouping of rs3798220 genotypes, comparing genotype - disease, genotype - serum LPA level, and deducing the correlation index of the relationship between LPA level and coronary heart disease, OELPA-CAD=1.503K 14.64, that is, the effect of coronary heart disease in serum LPA change K units. Conclusion: 1, LPA Gene: rs3798220 locus may affect coronary heart disease by affecting serum LPA level; rs7765803 locus may affect blood LPA level; 2, CLU gene: rs3216167 locus may affect serum LPA level and may be associated with coronary heart disease; rs7982 site may affect TC, LDL level, and rs1532278 locus may affect blood TC, levels, levels 3, LTF gene: rs1126477 locus may affect blood TG level 4, PLTP gene: rs394643 site may affect blood APOB level 5, serum LPA changes K unit history of coronary heart disease risk ORLPA-CAD=1.503K/14.64

【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：R541.4

【共引文獻(xiàn)】

相關(guān)期刊論文 前10條

1 祝小霞;曾勝煌;丁e醵，

本文編號：1853776