精神分裂症候选基因、临床症状及认知功能的相关性研究

发布时间：2018-07-26 19:54

【摘要】：精神分裂症是最常见、最严重的精神性疾病，多发于青壮年，世界人口中患病率约为1%；主要表现为精神活动与现实环境想脱离、认知、情感、意志及个性特征等各个方面不协调、相互分裂的疾病。精神分裂症发病主要受环境和遗传两大因素影响。家系、双生子及寄养子研究结果均表明遗传因素在其发病中具有重要作用。精神分裂症基因组扫描和候选基因研究，虽获得不少阳性结果，但重复性较差，结果争议很大。精神分裂症临床症状和认知功能受诸多因素影响，其遗传因素将会备受关注。目的利用生物信息学、分子遗传学技术及生物统计学方法，一方面探讨DBH、ACE、COMT、DRD2、CHRNA5、IL-10、IL-18、COX-2、TCF4、CACNA1C、MTHFR及GNB1L候选基因上SNPs与精神分裂症关联性，另一方面检验这些SNPs是否影响受试者临床症状和认知功能，试图揭示精神分裂症易感基因及影响临床症状和认知功能的遗传机制。方法本研究以350例首发精神分裂症，567例慢性精神分裂症和421例健康对照为研究对象。精神分裂症患者来自北京回龙观医院住院部，正常健康人群来自当地社区。所有研究对象均为中国汉族人，并且精神分裂症患者要符合ICD-10和CCMD-II-R的诊断标准。在征得受试者知情同意的情况下，采集外周静脉血，采用Promega (USA)液体纯化提取DNA试剂盒，提取基因组DNA。利用PCR-AFLP和SequenomMassArray技术，检测12个候选基因13个SNPs位点（rs141116007、rs4340、rs4680、rs1800497、rs3829787、rs1800872、rs1946518、rs689466、rs5275、rs2958182、rs2239050、rs1801133和rs748806）。应用在线遗传统计SHEsis软件计算基因型频率分布是否符合Hardy-Weinberg平衡定律；分析候选基因SNPs位点与精神分裂症的关联性；分析基因各个位点之间连锁不平衡程度及单倍型；应用MDR软件分析基因-基因间的交互作用；应用SPSS17.0软件分析候选基因SNPs位点与精神分裂症临床症状和认知功能的相关性。结果 1, H-W平衡检验和连锁不平衡程度分析（1）H-W平衡检验 rs4340、rs3829787和rs5275位点的基因型分布在首发精神分裂症组中偏离H-W平衡；rs1800497、rs3829787和rs1801133位点的基因型分布在慢性精神分裂症组中偏离H-W平衡；说明这些位点可能是疾病易感SNPs位点或与易感SNPs连锁；其他位点基因型分布在首发精神分裂症组、慢性精神分裂症组和健康对照组中均符合H-W平衡定律（all, P＞0.05），说明本研究抽样样本符合遗传学分析。（2）连锁不平衡程度分析 rs689466和rs5275处于高度连锁不平衡染色体区，说明其位于连锁不平衡区域内。 2，病例-对照分析 (1)首发精神分裂症-正常对照组单个SNP分析 rs141116007和rs5275位点与精神分裂症发病相关联，其等位基因和基因型频率分布在病例组和对照组中均出现显著性差异（all, P0.05）。rs2239050和rs3829787位点可能与精神分裂症发病相关联。rs2239050等位基因和rs3829787基因型的频率分布在病例组和对照组中出现显著差异（both, P0.05）。 (2)慢性精神分裂症-正常对照组单个SNP分析 rs4340、rs1801133和rs748806位点与精神分裂症发病相关联，其等位基因和基因型频率分布在两组中均出现显著性差异（all, P0.05）。rs1800497位点可能与精神分裂症发病相关联，其基因型频率分布在病例组和对照组中出现显著差异（P0.05）。 (3)病例对照组单倍型分析对COX-2基因的2个SNPs位点进行联合分析。结果显示，仅rs689466(C)-rs5275(A)作为保护型单倍型与首发精神分裂症相关联（P0.05）。 3，SNPs与精神分裂症临床症状的相关分析 (1) SNPs与首发精神分裂症临床症状的相关分析在首发精神分裂症中，DBH基因rs141116007位点和IL18基因rs1946518位点与首发精神分裂症阳性症状和临床症状总分相关（all, P0.05）；ACE基因rs4340位点与首发精神分裂症阴性症状相关（P0.05）。 (2) SNPs与慢性精神分裂症临床症状的相关分析在慢性精神分裂症中，COX-2基因rs689466和rs5275位点与慢性精神分裂症的阳性症状相关（both, P0.05）；TCF4基因rs2958182位点与慢性精神分裂症阴性症状相关（P0.05）；IL-18基因rs1946518位点与慢性精神分裂症临床症状总分相关（P0.05）。 4，SNPs与认知功能的相关分析 (1) SNPs与正常健康人认知功能的相关分析在正常健康对照中，TCF4基因rs2958182和CACNA1C基因rs2239050位点与正常健康人认知功能的注意相关（both, P0.05）；COX-2基因rs5275位点与正常健康人认知功能的语言相关（P0.05）；TCF4基因rs2958182位点与正常健康人认知功能的延迟记忆和总分值相关（both, P0.05）。 (2) SNPs与首发精神分裂症认知功能的相关分析在首发精神分裂症中，DBH基因rs141116007、DRD2基因rs1800497位点和COX-2基因rs5275位点与首发精神分裂症认知功能的即刻记忆相关（all,P0.05）；COX-2基因rs689466位点和rs5275位点与首发精神分裂症认知功能的语言相关（all, P0.05）。 (3) SNPs与慢性精神分裂症认知功能的相关分析在慢性精神分裂症中，MTHFR基因rs18001133位点和IL-10基因rs1800872位点与慢性精神分裂症注意能力相关（both, P0.05）；TCF4基因rs2958182位点和MTHFR基因rs18001133位点与慢性精神分裂症的语言能力相关（both,P0.05）；DRD2基因rs1800497位点和TCF4基因rs2958182位点与慢性精神分裂症认知功能的延迟记忆相关（both, P0.05）；TCF4基因rs2958182位点与慢性精神分裂症认知功能的总分值相关（P0.05）。结论从上述分析可得到如下结论：(1) DBH、COX-2、CHRNA5和CACAN1C基因可能是首发精神分裂症的易感基因；（2）ACE、DRD2、MTHFR和GNB1L基因可能是慢性精神分裂症的易感基因；(3)首发精神分裂症中DBH、IL-18基因和慢性精神分裂症中COX-2基因分别与阳性症状相关（；4）首发精神分裂症中ACE基因和慢性精神分裂症中TCF4基因分别与阴性症状相关；（5）首发精神分裂症中DBH、IL-18基因和慢性精神分裂症中IL-18基因分别与临床症状总分相关；（6）TCF4、CACNA1C和COX-2基因与正常健康人群认知功能功能相关；（7）DBH、DRD2和COX-2基因与首发精神分裂症认知功能相关（；8）MTHFR、IL-10、DRD2、TCF4基因与慢性精神分裂症认知功能相关。（9）精神分裂症存在遗传和临床的异质性。
[Abstract]:Schizophrenia is the most common and most serious mental disease, mostly in young and young adults. The prevalence rate of the world population is about 1%. The main manifestations are the disharmony between mental activity and the realistic environment, the disharmony of cognition, emotion, will and personality. The main causes of schizophrenia are two major causes of environment and heredity. The results of the study of the family, the twins and the mailing seed all showed that the genetic factors played an important role in the disease. The genome scan and candidate gene study of schizophrenia have obtained a lot of positive results, but the reproducibility is poor and the result is very controversial. The clinical symptoms and cognitive functions of schizophrenia are affected by many factors, and their genetic causes are affected. It's going to get a lot of attention.
objective
Using bioinformatics, molecular genetics and biometric methods, the association of DBH, ACE, COMT, DRD2, CHRNA5, IL-10, IL-18, COX-2, TCF4, CACNA1C, MTHFR and GNB1L candidate genes to schizophrenia, on the other hand, to examine whether these effects affect the clinical symptoms and cognitive functions of the subjects and try to reveal the mental points. Susceptibility genes and genetic mechanisms affecting clinical symptoms and cognitive functions.
Method
In this study, 350 cases of first onset schizophrenia, 567 chronic schizophrenia and 421 healthy controls were studied. Schizophrenic patients came from the hospitalized Department of Beijing Hui Long Guan Hospital, and the normal healthy population came from the local community. All the subjects were Chinese Han people, and the patients with seminal schizophrenia had to meet the ICD-10 and CCMD-II-R diagnosis. Break the standard.
In the case of informed consent of the subjects, the peripheral venous blood was collected and the DNA kit was extracted with Promega (USA) liquid, and the genomic DNA. was extracted by PCR-AFLP and SequenomMassArray technology to detect the 13 SNPs loci of the 12 candidate genes (rs141116007, rs4340, rs4680, rs1800497, rs3829787. 958182, rs2239050, rs1801133 and rs748806).
The application of online genetic statistical SHEsis software to calculate whether the genotype frequency distribution is consistent with the Hardy-Weinberg equilibrium law; analyze the correlation between the candidate gene SNPs locus and schizophrenia; analyze the linkage disequilibrium and haplotypes between the various loci of the gene; use the MDR software to analyze the interaction between the gene and the gene, and the application of SPSS17. 0 software analysis of the correlation between the candidate gene SNPs locus and clinical symptoms and cognitive function of schizophrenia.
1, H-W balance test and linkage disequilibrium analysis
(1) H-W balance test
The genotype distribution of the rs4340, rs3829787 and rs5275 loci in the first schizophrenic group deviated from the H-W balance; the genotype distribution of the rs1800497, rs3829787 and rs1801133 loci was deviated from the H-W balance in the chronic schizophrenia group, indicating that these loci may be susceptible to the disease, or to susceptible SNPs, and other loci genotypes. The cloth was consistent with the H-W equilibrium law (all, P > 0.05) in the first schizophrenic group, the chronic schizophrenia group and the healthy control group, indicating that the sample samples in this study were in accordance with the genetic analysis.
(2) analysis of the degree of linkage disequilibrium
Rs689466 and rs5275 are in highly unbalanced chromosomal regions, indicating that they are located in the linkage disequilibrium region.
2, case control analysis
(1) Single SNP analysis in first-episode schizophrenia-normal control group
Rs141116007 and rs5275 loci were associated with schizophrenia. The frequency distribution of alleles and genotypes in the case group and the control group were significantly different (all, P0.05).Rs2239050 and rs3829787 sites may be associated with schizophrenia, the frequency of.Rs2239050 alleles and the frequency of rs3829787 genotypes in the case group There was a significant difference between the control group and the control group (both, P0.05).
(2) Single SNP analysis in chronic schizophrenia-normal control group
Rs4340, rs1801133 and rs748806 loci were associated with schizophrenia. The allele and genotype frequency distribution in the two groups showed significant differences (all, P0.05).Rs1800497 locus may be associated with schizophrenia, and its genotype frequency distribution was significantly different in the case group and the control group (P0.05).
(3) haplotype analysis in case control group
The combined analysis of 2 SNPs loci of the COX-2 gene showed that only rs689466 (C) -rs5275 (A) as a protective haplotype was associated with first episode schizophrenia (P0.05).
3. Correlation between SNPs and clinical symptoms of schizophrenia
(1) correlation analysis between SNPs and clinical symptoms of first-episode schizophrenia
In the first episode of schizophrenia, the rs141116007 locus of DBH gene and the rs1946518 locus of the IL18 gene were related to the positive symptoms of first schizophrenia and the total score of clinical symptoms (all, P0.05), and the rs4340 site of the ACE gene was associated with the negative symptoms of first episode schizophrenia (P0.05).
(2) correlation analysis between SNPs and clinical symptoms of chronic schizophrenia
In chronic schizophrenia, the COX-2 gene rs689466 and rs5275 loci are related to the positive symptoms of chronic schizophrenia (both, P0.05); the rs2958182 locus of the TCF4 gene is associated with the negative symptoms of chronic schizophrenia (P0.05), and the IL-18 gene rs1946518 locus is associated with the total score of the clinical symptoms of chronic schizophrenia (P0.05).
4, SNPs and the correlation analysis of cognitive function
(1) Correlation between SNPs and Cognitive Function of Normal Healthy Persons
In normal healthy controls, the rs2239050 loci of the TCF4 gene rs2958182 and CACNA1C genes are related to the cognitive function of normal healthy people (both, P0.05); the COX-2 gene rs5275 loci is related to the language of normal healthy people (P0.05); the rs2958182 locus of the TCF4 gene is associated with the delayed memory and total score of cognitive function of normal healthy people. Both, P0.05.
(2) correlation between SNPs and cognitive function in first-episode schizophrenia
In the first episode of schizophrenia, the DBH gene rs141116007, the rs1800497 locus of the DRD2 gene, and the rs5275 locus of the COX-2 gene are associated with the immediate memory of the cognitive function of the first schizophrenia (all, P0.05), and the rs689466 and rs5275 loci of the COX-2 gene are related to the language of the cognitive ability of first episode schizophrenia.
(3) correlation between SNPs and cognitive function in chronic schizophrenia
In chronic schizophrenia, the rs18001133 locus of the MTHFR gene and the rs1800872 locus of the IL-10 gene are related to the attention ability of chronic schizophrenia (both, P0.05), and the TCF4 gene rs2958182 site and the MTHFR gene rs18001133 locus are related to the language ability of chronic schizophrenia (both, P0.05); The 58182 locus is associated with delayed memory of cognitive function of chronic schizophrenia (both, P0.05), and the rs2958182 locus of the TCF4 gene is associated with the total score of cognitive function of chronic schizophrenia (P0.05).
conclusion
From the above analysis, we can get the following conclusions: (1) DBH, COX-2, CHRNA5 and CACAN1C genes may be the susceptible genes of first onset schizophrenia; (2) ACE, DRD2, MTHFR and GNB1L genes may be the susceptible genes of chronic schizophrenia; (3) the COX-2 genes in DBH, IL-18 genes and chronic schizophrenia in first episode schizophrenia are respectively and positive. Symptoms related (; 4) the TCF4 gene of ACE gene and chronic schizophrenia in first episode schizophrenia was associated with negative symptoms respectively. (5) the IL-18 gene of DBH, IL-18 and chronic schizophrenia in first episode schizophrenia was related to the total score of clinical symptoms, and (6) TCF4, CACNA1C and COX-2 genes and the cognitive function work of normal healthy people. Can be related; (7) DBH, DRD2 and COX-2 genes are associated with cognitive function of first episode schizophrenia (; 8) MTHFR, IL-10, DRD2, TCF4 genes are associated with cognitive function of chronic schizophrenia. (9) there is genetic and clinical heterogeneity in schizophrenia.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2014
【分类号】：R749.3

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