中国汉族人群MVP、AQP4、KCNJ10基因多态与部分性癫痫耐药的关联研究

发布时间：2018-03-28 20:18

本文选题：部分性癫痫　切入点：耐药　出处：《中南大学》2014年博士论文

【摘要】：研究目的：癫痫(epilepsy, EP)是神经系统常见的慢性疾病之一,其中有30%左右的癫痫患者在经过合理选用的抗癫痫药物(antiepileptic drugs, AEDs)治疗之后,仍不能有效控制发作,发展成为耐药性癫痫,给患者及家人带来沉重负担。耐药性癫痫的形成被认为是环境与遗传因素共同作用的结果。近年来研究提示,主穹窿蛋白(major vault protein, MVP)、水通道蛋白4(aquaporin4, AQP4)以及内向整流K+离子通道Kir4.1可能参与了癫痫的耐药过程。癫痫的耐药可能与基因单核苷酸多态(single nucleotide polymorphisms, SNPs)在不同地区的种族人群中存在关联。本课题拟在中国汉族人群中探讨MVP、AOP4、KCNJ10基因SNPs与部分性癫痫耐药性是否相关,为部分性癫痫的药物个体化治疗提供新的思路。研究方法：依据国际抗癫痫联盟(International League Against Epilepsy, ILAE)对癫痫分类和耐药性癫痫的诊断标准,于中南大学湘雅医院神经内科癫痫专科门诊收集部分性癫痫病例510例,其中222例为耐药组,288例为非耐药组,配对选取健康人群对照206例,研究对象均为汉族,获得知情同意后详细询问癫痫病史并进行临床评估,抽取外周静脉血提取全基因组DNA。从国际人类基因组单体型图Hapmap数据库通过Haploview (version4.2)软件对中国汉族人群MVP、AOP4\KCNJ10基因SNP位点进行筛选。应用Sequenom MassARRAY SNP检测方法对候选SNP位点进行基因分型。比较MVP、KCNJ10、AQP4基因SNP位点基因型频率、等位基因频率、连锁不平衡(linkage disequilibrium, LD)及单倍型频率在部分性癫痫耐药组与非耐药组、部分性癫痫与健康对照组间的差异,分析MVP、AOP4、KCNJ10基因SNPs是否在中国汉族人群中与部分性癫痫耐药存在关联。研究结果： 1.部分性癫痫耐药组相对于非耐药组,发病起始年龄较早,服药前的癫痫发作频率相对较高,癫痫病程相对较长(P0.05)； 2.MVP基因4个SNP位点基因型频率、等位基因频率在部分性癫痫耐药组与非耐药组、部分性癫痫组与健康对照组之间未见明显差异(P0.05)； 3.AQP4基因3个SNP位点、KCNJ10基因9个SNP位点的基因型频率、等位基因频率在部分性癫痫耐药组与非耐药组间未见明显差异(P0.05),在特发部分性癫痫组与健康对照组间,KCNJ10rs1890532CG基因型在健康人群中频率高于癫痫人群(OR=0.636,95%CI (0.409-0.989), P=0.045);但经Bofferoni方法进行多重检验校正后P=0.405,组间比较未见明显差异； 4)部分性癫痫耐药组与非耐药组在SNP位点LD分析中,MVP基因rs12149746、 rs1057451、rs9938630,AQP4基因rs1058424、 rs3763043、rs35931以及KCNJ10基因rs12122979、rs1186685.rs6690889位点间分别存在LD并形成单倍域(D’0.7且r20.4),组间单倍型频率分析未见明显差异(P0.05) 研究结论： 1.部分性癫痫发病年龄、服药前发作频率、病程长短可作为耐药的风险评估指标； 2.MVP基因SNPs在中国汉族人群中可能与部分性癫痫的耐药及易感性不相关； 3.AQP4基因和KCNJ10基因SNPs在中国汉族人群中可能与部分性癫痫的耐药及易感性不相关。 5. MVP、AQP4、KCNJ10基因SNP位点单倍型在中国汉族人群中可能与部分性癫痫的耐药不相关。
[Abstract]:Objective: epilepsy (epilepsy, EP) is one of the common chronic diseases of the nervous system, which has about 30% patients with epilepsy after antiepileptic drug selection (antiepileptic drugs, AEDs) after treatment, still can not effectively control the seizures, become resistant epilepsy, bring heavy burden to patients and their families. The formation of drug resistance epilepsy is considered the interaction of environmental and genetic factors. Recent studies suggest that major vault protein (major vault protein, MVP), aquaporin 4 (aquaporin4, AQP4) and inward rectifier K+ channels may be involved in the process of Kir4.1 resistant epilepsy. Epilepsy drug resistance and gene single nucleotide polymorphisms (single nucleotide polymorphisms, SNPs) association exists in different regions of the ethnic groups. In this study Chinese Han population of MVP, AOP4, SNPs and partial KCNJ10 gene Whether the drug resistance is associated with epilepsy provides a new way of thinking for the individualized treatment of drugs in partial epilepsy.
Research methods: according to the International League Against Epilepsy (International League Against Epilepsy, ILAE) on seizure classification and drug resistance epilepsy diagnosis standard in Xiangya Hospital of Central South University Department of neurology clinic collected partial epilepsy 510 cases, including 222 cases of drug resistant group, 288 cases of non resistance group, 206 normal subjects were chosen the object of study, all were Han nationality, after obtaining informed consent and asked in detail about the history of epilepsy and clinical assessment, extraction of genomic DNA. from the International HapMap database through the Hapmap Haploview peripheral venous blood samples (version4.2) software to China Han population MVP AOP4KCNJ10 SNP gene were selected. Using Sequenom MassARRAY SNP detection method. Genotyping of the candidate SNP sites. MVP, KCNJ10, AQP4 gene SNP genotype, allele frequencies, linkage The difference between linkage disequilibrium (LD) and haplotype frequency in partial epilepsy drug resistant group and non drug resistant group was between partial epilepsy and healthy control group. Whether MVP, AOP4 and KCNJ10 gene SNPs were associated with partial epilepsy drug resistance in Chinese Han population were analyzed.
The results of the study:
In the 1. part, the onset age of the epileptic drug resistance group was earlier than that of the non drug resistant group. The frequency of epileptic seizures before treatment was relatively high, and the duration of epilepsy was relatively long (P0.05).
The genotype frequencies and allele frequencies of 4 SNP loci in 2.MVP gene were not significantly different between the partial epilepsy group and the non drug resistant group. There was no significant difference between the partial epilepsy group and the healthy control group (P0.05).
3 SNP loci of 3.AQP4 gene, the genotype frequencies of 9 SNP loci of KCNJ10 gene, allele frequency in partial epilepsy resistant group and non resistance group, there was no significant difference between (P0.05), in idiopathic partial epilepsy group and the healthy control group, KCNJ10rs1890532CG genotype in the healthy population is higher than the frequency of epileptic patients (OR=0.636,95%CI (0.409-0.989), P=0.045); but the Bonferroni correction by P=0.405 Bofferoni method, comparison between groups showed no significant difference;
4) partial epilepsy resistant group and non resistance group analysis in the SNP locus in LD gene, MVP rs12149746, rs1057451, rs9938630, AQP4, rs3763043, rs35931, rs1058424 gene, KCNJ10 gene rs12122979, rs1186685.rs6690889 locus exists respectively and the formation of LD single domain (D 0.7 and r20.4), there was no significant difference between the analysis of haplotype frequencies between groups (P0.05)
The conclusions are as follows:
The age of the 1. sexual epilepsy, the frequency of the attack before taking medicine, the length of the disease can be used as the risk assessment index of the drug resistance.
The 2.MVP gene SNPs may not be related to the resistance and susceptibility of partial epilepsy in Chinese Han population.
The 3.AQP4 gene and the KCNJ10 gene SNPs may not be associated with resistance and susceptibility to partial epilepsy in Chinese Han population.
The haplotype of 5. MVP, AQP4, and KCNJ10 gene SNP loci may not be related to the resistance of partial epilepsy in Chinese Han population.

【学位授予单位】：中南大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R742.1

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