氯吡格雷抗血小板效应及药动学的全基因组关联性研究
发布时间：2018-10-19 09:31
【摘要】：氯吡格雷是冠心病治疗的一线抗血小板药物,能有效降低经皮冠状动脉介入治疗(PCI)后冠心病患者的死亡率和再缺血事件的发生。但其疗效存在较大个体差异,而目前大部分个体差异仍然无法解释。氯吡格雷需在体内代谢转化为活性代谢物H4才能发挥抗血小板聚集作用,所以涉及其代谢活化的基因多态性可能对不同个体间疗效的差异有重要影响。因此,本研究拟结合抗血小板效应和药动学过程进行全基因组关联性分析(GWAS),系统发现影响中国冠心病患者氯吡格雷疗效的新功能性基因变异,并评价其对主要不良心血管事件(MACE)发生的预测作用。首先,在115名患者中检测服药后血小板集聚率(PRU),进行GWAS分析。同时考察H4血药浓度及特别关注药动学通路的基因,筛选出18个可能影响氯吡格雷抗血小板效应和药动学的候选SNP位点。进一步研究候选SNP位点与氯吡格雷药动学的相关性。入选31名患者作为研究对象,给予氯吡格雷后进行药动学考察及候选位点分型检测,分析候选位点基因型与氯吡格雷和H4药动学参数(Cmax、Tmax、AUC0-4h)的关联性。除了CYP2C19*2外,N6AMT1 rs2254638、SLC14A2 rs12456693和ABC 1 rs2487032也与氯吡格雷的体内药动学过程显著相关。接着考察候选位点是否对氯吡格雷的活化代谢有潜在影响。收集32名患者正常肝组织,制备肝S9组分。建立氯吡格雷在人肝S9的体外孵育体系,分析在各个体S9中代谢物的生成量。检测候选位点基因分型,分析不同基因型对氯吡格雷代谢的影响。发现CYP2C19*2及N6AMT1rs2254638与H4的生成显著相关。结合上述结果发现,N6AMT1 rs2254638和ABCA1 rs2487032是PRU的独立影响因素,可极大提高对PRU变异的解释程度至37.7%,而目前文献报道只有约20%;N6AMT1 rs2254638和SLC14A2rs12456693是H4血药浓度的独立预测因子。最后,评价候选位点对接受氯吡格雷治疗的冠心病患者MACE事件发生的预测作用,探寻基因多态性是否可作为与终点事件相关的遗传标记。入选患者为前瞻性研究队列,并进行定期随访。将PCI术后1.5年内发生MACE事件的91名患者,作为病例组;另外随机抽取208名患者,作为对照组。检测候选位点分型,运用Logistic回归模型分析,发现N6AMT1 rs2254638、SLC14A2 rs12456693和ATP10Ars12913988可增加MACE事件发生的风险。推测位于N6AMT1和SLC14A2的位点是通过影响氯毗格雷及H4在体内的药动学过程从而影响其抗血小板效应。综上所述,本研究系统发现了新的影响氯吡格雷疗效的基因变异,极大提高对其反应性差异的解释程度,以及影响其疗效的吸收、代谢机制的理解;有助于氯吡格雷治疗的个体化用药。
[Abstract]:Clopidogrel is a first-line antiplatelet drug for coronary heart disease, which can effectively reduce the mortality and reischemic events in patients with coronary heart disease after percutaneous coronary intervention (PCI). However, there are large individual differences in the efficacy, and most of the individual differences are still unexplained. Clopidogrel needs to be metabolized into active metabolite H4 in vivo in order to play an anti-platelet aggregation effect, so the gene polymorphism involved in its metabolic activation may have an important effect on the difference of efficacy among individuals. Therefore, this study was designed to combine the antiplatelet effect and pharmacokinetic process to analyze the genome-wide association of Chinese coronary heart disease patients with clopidogrel by (GWAS), system and to identify new functional gene mutations that affect the efficacy of clopidogrel in Chinese patients with coronary heart disease. To evaluate its predictive effect on the occurrence of major adverse cardiovascular events (MACE). First, the platelet aggregation rate (PRU),) was analyzed by GWAS in 115 patients. At the same time, the blood concentration of H4 and the genes of pharmacokinetic pathway were studied. Eighteen candidate SNP sites were screened out which may affect the antiplatelet effect and pharmacokinetics of clopidogrel. To further study the correlation between candidate SNP sites and clopidogrel pharmacokinetics. The pharmacokinetics and candidate locus typing of clopidogrel were investigated in 31 patients. The correlation between candidate locus genotypes and clopidogrel and H4 pharmacokinetic parameters (Cmax,Tmax,AUC0-4h) was analyzed. In addition to CYP2C19*2, N6AMT1 rs2254638,SLC14A2 rs12456693 and ABC 1 rs2487032 were also significantly associated with clopidogrel's pharmacokinetics in vivo. Then we investigated whether the candidate sites had a potential effect on the activation and metabolism of clopidogrel. The normal liver tissues of 32 patients were collected and liver S 9 components were prepared. The in vitro incubation system of clopidogrel in human liver S9 was established and the amount of metabolites produced in each individual S9 was analyzed. The genotypes of candidate loci were detected and the effects of different genotypes on clopidogrel metabolism were analyzed. It was found that CYP2C19*2 and N6AMT1rs2254638 were significantly correlated with H4 production. Combined with the above results, it was found that N6AMT1 rs2254638 and ABCA1 rs2487032 were independent influencing factors of PRU and could greatly improve the degree of interpretation of PRU variation to 37.70.AMT 1 rs2254638 and SLC14A2rs12456693 were only reported to be independent predictors of serum concentration of H4 by about 20% N6AMT1 rs2254638 and SLC14A2rs12456693. Finally, we evaluated the predictive effect of candidate sites on the occurrence of MACE events in patients with coronary heart disease treated with clopidogrel, and explored whether gene polymorphism could be used as a genetic marker associated with endpoint events. The patients were enrolled in a prospective study cohort and were followed up regularly. 91 patients with MACE events within 1.5 years after PCI were selected as case group, and another 208 patients were randomly selected as control group. By using Logistic regression model, we found that N6AMT1 rs2254638,SLC14A2 rs12456693 and ATP10Ars12913988 can increase the risk of MACE events. It is assumed that the sites located in N6AMT1 and SLC14A2 affect the antiplatelet effect by affecting the pharmacokinetic process of clopidogrel and H4 in vivo. To sum up, we found new gene variations that affect the efficacy of clopidogrel, greatly improve the degree of explanation of the difference in its reactivity, and influence the absorption and metabolic mechanism of clopidogrel. It is helpful for the treatment of clopidogrel.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R969
本文编号：2280746
[Abstract]:Clopidogrel is a first-line antiplatelet drug for coronary heart disease, which can effectively reduce the mortality and reischemic events in patients with coronary heart disease after percutaneous coronary intervention (PCI). However, there are large individual differences in the efficacy, and most of the individual differences are still unexplained. Clopidogrel needs to be metabolized into active metabolite H4 in vivo in order to play an anti-platelet aggregation effect, so the gene polymorphism involved in its metabolic activation may have an important effect on the difference of efficacy among individuals. Therefore, this study was designed to combine the antiplatelet effect and pharmacokinetic process to analyze the genome-wide association of Chinese coronary heart disease patients with clopidogrel by (GWAS), system and to identify new functional gene mutations that affect the efficacy of clopidogrel in Chinese patients with coronary heart disease. To evaluate its predictive effect on the occurrence of major adverse cardiovascular events (MACE). First, the platelet aggregation rate (PRU),) was analyzed by GWAS in 115 patients. At the same time, the blood concentration of H4 and the genes of pharmacokinetic pathway were studied. Eighteen candidate SNP sites were screened out which may affect the antiplatelet effect and pharmacokinetics of clopidogrel. To further study the correlation between candidate SNP sites and clopidogrel pharmacokinetics. The pharmacokinetics and candidate locus typing of clopidogrel were investigated in 31 patients. The correlation between candidate locus genotypes and clopidogrel and H4 pharmacokinetic parameters (Cmax,Tmax,AUC0-4h) was analyzed. In addition to CYP2C19*2, N6AMT1 rs2254638,SLC14A2 rs12456693 and ABC 1 rs2487032 were also significantly associated with clopidogrel's pharmacokinetics in vivo. Then we investigated whether the candidate sites had a potential effect on the activation and metabolism of clopidogrel. The normal liver tissues of 32 patients were collected and liver S 9 components were prepared. The in vitro incubation system of clopidogrel in human liver S9 was established and the amount of metabolites produced in each individual S9 was analyzed. The genotypes of candidate loci were detected and the effects of different genotypes on clopidogrel metabolism were analyzed. It was found that CYP2C19*2 and N6AMT1rs2254638 were significantly correlated with H4 production. Combined with the above results, it was found that N6AMT1 rs2254638 and ABCA1 rs2487032 were independent influencing factors of PRU and could greatly improve the degree of interpretation of PRU variation to 37.70.AMT 1 rs2254638 and SLC14A2rs12456693 were only reported to be independent predictors of serum concentration of H4 by about 20% N6AMT1 rs2254638 and SLC14A2rs12456693. Finally, we evaluated the predictive effect of candidate sites on the occurrence of MACE events in patients with coronary heart disease treated with clopidogrel, and explored whether gene polymorphism could be used as a genetic marker associated with endpoint events. The patients were enrolled in a prospective study cohort and were followed up regularly. 91 patients with MACE events within 1.5 years after PCI were selected as case group, and another 208 patients were randomly selected as control group. By using Logistic regression model, we found that N6AMT1 rs2254638,SLC14A2 rs12456693 and ATP10Ars12913988 can increase the risk of MACE events. It is assumed that the sites located in N6AMT1 and SLC14A2 affect the antiplatelet effect by affecting the pharmacokinetic process of clopidogrel and H4 in vivo. To sum up, we found new gene variations that affect the efficacy of clopidogrel, greatly improve the degree of explanation of the difference in its reactivity, and influence the absorption and metabolic mechanism of clopidogrel. It is helpful for the treatment of clopidogrel.
【学位授予单位】：南方医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R969
【参考文献】
相关期刊论文 前2条
1 钟诗龙;韩雅玲;陈纪言;袁晋青;孙艺红;王效增;李毅;;氯吡格雷抗血小板治疗个体化用药基因型检测指南解读[J];中国实用内科杂志;2015年01期
2 康彦红;劳海燕;余细勇;陈纪言;钟诗龙;;经皮冠状动脉介入治疗术后支架内再狭窄的遗传学与表观遗传学研究进展[J];中华医学遗传学杂志;2012年01期
，本文编号：2280746
本文链接：https://www.wllwen.com/yixuelunwen/yiyaoxuelunwen/2280746.html
最近更新
教材专著
