Peli-1基因单核苷酸多态性及其mRNA水平与系统性红斑狼疮的相关性研究
发布时间：2018-07-16 18:10
【摘要】：研究背景系统性红斑狼疮(Systemic lupus erythematosus,SLE),是一种好发于女性的慢性自身免疫性疾病,其临床表现较为复杂,病程迁延反复,常常累及多个系统和器官。近年来,国内外大量研究致力于探索SLE的病因和发病机制,但其确切的机制尚未完全明了。目前一般认为其发病涉及遗传因素、环境因素、内分泌状况等多种因素,这些因素相互影响、相互作用,构成一个复杂的病因网,引起异常免疫反应,如炎症因子释放、自身抗体产生。SLE最初被认为是B细胞介导的自身免疫性疾病。随着研究的深入,T细胞被发现在SLE的发生、发展过程中也起着至关重要的作用。1999年科学家首次在果蝇体内发现一种高度保守的E3泛素化连接酶蛋白,命名为Pellino(简称Peli)蛋白。该种泛素化连接酶蛋白家族一共有Peli-1、Peli-2、Peli-3三种亚型,其最值得一提的功能特征是在先天性免疫中承担多元性角色。最新研究发现Peli-1在参与TLR/IL-1R介导的一系列细胞信号传导起着一定的调节作用。Peli-1还参与介导TRIF依赖的NF-κB活化。此外,Chang等发现Peli-1是维持T细胞耐受的一个关键调节因子,其功能异常可能导致T细胞激活,从而可能在SLE等自身免疫性疾病的发病中起重要作用。基于以上内容,本课题首次探讨Peli-1基因多态性与SLE的疾病易感性之间的关联;同时检测Peli-1 mRNA表达水平及其与SLE易感性和SLE不同表型之间的相关性,从基因转录、表达水平分别探讨Peli-1基因与SLE发病之间的关系。第一部分peli-1基因单核苷酸多态性与系统性红斑狼疮的相关性研究目的比较sle患者于正常对照peli-1rs329498、rs10496105等位基因和基因频率的差异,探讨其与sle遗传易感性的关联。并结合临床资料,分析peli-1rs329498、rs10496105与其主要临床特征及其实验室指标之间的关系。方法采用病例对照的研究方法,选取738名sle患者和827名健康对照。采用fluidigm192.24基因分型技术检测peli-1基因中rs329498、rs10496105两个snp位点,对其基因型、等位基因频率等进行分析,并结合临床资料进一步分析基因频率分布于临床表现之间的关系。结果(1)peli-1基因rs329498位点多态性与sle的关联性研究sle患者组aa、ac、cc的基因型频率分别为40.8%、44.6%、14.6%,对照组分别为36.5%、45.5%、18.0%,两组差异无统计学意义(χ2=4.63,p=0.099)。等位基因比较显示两组差异有统计学意义(χ2=4.80,p=0.028),相对大等位基因a,小等位基因c会降低sle发病的风险(or=0.851,95%ci:0.737-0.983)。但遗传显性、隐性模型均未见统计学关联(均有p0.05)。与非ln肾炎组患者相比,ln患者a等位基因频率较高,且差异有统计学意义(χ2=8.18,p=0.017)。小等位基因c相比于a等位基因,其ln发病风险or值为0.681(95%ci:0.528-0.880)。两组显性模型基因型频率(cc+acversusaa)分布差异有统计学意义(or=0.632,95%ci:0.451-0.884,p=0.007)。临床特征关联性分析显示peli-1基因rs329498位点可能与颧部红斑有关,其基因型频率分布有统计学差异(χ2=6.63,p=0.036)。(2)peli-1基因rs10496105位点多态性与sle的关联性研究位点rs10496105基因型频率gg、ag、aa在病例组分别为70.0%、26.2%和3.8%,对照组中分别为69.4%,27.2%和3.4%。病例组与对照组基因型频率分布无统计学差异(χ2=0.37,p=0.832)。遗传模型分析结果表明,无论是显性模型还是隐性模型病例组和对照组分布均未发现统计学差异(均有p0.05)。此外,ln组与非LN组间基因型分布差异无统计学意义(χ2=3.11,P=0.201)。临床资料分析结果显示rs10496105位点与颧部红斑、盘状红斑、光敏感、口腔溃疡、关节炎等主要临床特征也无统计学关联(均有P0.05)。结论中国汉族人群中Peli-1 rs329498基因多态性与SLE遗传易感性有一定关联,此外,rs329498位点也与狼疮肾炎及颧部红斑有关。而未发现rs10496105基因多态性位点与SLE之间的统计学关联。第二部分Peli-1 m RNA水平与系统性红斑狼疮的相关性研究目的分别比较SLE患者组和正常对照组、SLE患者中狼疮肾炎组与非狼肾炎组以及活动组与非活动组外周血单个核细胞(Peripheral blood mononuclear cells,PBMCs)中Peil-1基因m RNA表达水平,并探讨其表达水平与临床特征、实验室指标以及疾病活动度之间的关系。方法共收集31例SLE患者和30例健康对照。采用Trizol法提取PBMCs中的RNA。实时定量聚合酶链反应(Real time quantitative polymerase chain reaction,RT-PCR)检测PBMCs中的Peli-1基因m RNA表达水平,利用Mann-Whitney秩和检验比较各组之间m RNA水平是否存在差异。采用Spearman相关分析Peli-1基因m RNA表达水平与系统性红斑狼疮疾病活动指数(Systemic lupus erythematosus disease activity index,SLEDAI)关系。所有数据均采用SPSS 17.0进行统计分析。结果(1)PBMCs中Peli-1 m RNA表达水平比较:31例SLE患者与30例对照组PBMCs中的Peli-1 m RNA水平无统计学差异(Z=-0.404,P=0.686)。将SLE患者分为肾炎组(11例)和非肾炎组(20例)比较,结果显示两组PBMCs中的Peli-1m RNA水平无统计学差异(Z=-0.619,P=0.536)。此外,活动组(19例)和非活动组(12例)组间的PBMCs中的Peli-1 m RNA水平无统计学差异(Z=-0.973,P=0.330)。(2)SLE患者PBMCs中Peli-1 m RNA水平与其临床特征关联性分析:Mann-Whitney秩和检验结果显示,各组之间Peli-1 m RNA水平差异均无统计学意义(均有P0.05)。(3)SLE患者PBMCs中Peli-1 m RNA水平与实验室指标关联性分析:各个实验室指标阳性组与阴性组间Peli-1 m RNA水平差异均无统计学意义(均有P0.05)。(4)SLE患者PBMCs中Peli-1 m RNA表达水平与SLEDAI相关性分析:Spearman等级相关分析结果显示,SLE患者PBMCs中Peli-1 m RNA水平与SLEDAI评分之间无明显相关性(rs=0.148,P=0.428)。结论本研究发现SLE患者与健康对照组、患者活动组与非活动组、LN与非LN组之间Peli-1基因m RNA表达水平无统计学差异。此外,通过分析比较各主要临床表现及实验室主要特征,结果也均未发现统计学差异。SLEDAI评分与Peli-1基因m RNA表达水平也无明显相关性。
[Abstract]:Research background systemic lupus erythematosus (Systemic lupus erythematosus, SLE) is a kind of chronic autoimmune disease in women. Its clinical manifestation is more complex, the course of disease is prolonged, and many systems and organs are often involved. In recent years, a lot of studies have been made to explore the etiology and pathogenesis of SLE, but the exact mechanism of it has been studied. It is not yet fully understood. It is generally considered to be associated with a variety of factors such as genetic, environmental and endocrine factors that interact and interact to form a complex network of etiological factors that cause abnormal immune responses, such as the release of inflammatory factors, and the production of.SLE by autoantibodies is initially considered to be an autoimmune disease mediated by B cells. Disease. With the development of research, T cells have been found to occur in SLE and also play a vital role in the development of.1999. A highly conserved E3 ubiquitin ligase protein, named Pellino (abbreviated Peli) protein, was found in the fruit fly for the first time. The ubiquitin ligase protein family consists of Peli-1, Peli-2, Peli-3 three. The most notable functional feature of the subtype is the multiple role in innate immunity. The latest research has found that Peli-1 plays a regulatory role in a series of cellular signaling mediated by TLR/IL-1R,.Peli-1 also participates in the activation of NF- kappa B, which is mediated by TRIF dependence. In addition, Chang has found that Peli-1 is one of the maintenance of T cell tolerance. A key regulatory factor, whose functional abnormalities may lead to T cell activation, may play an important role in the pathogenesis of autoimmune diseases such as SLE. Based on the above, this subject is the first to explore the association between the Peli-1 gene polymorphism and the susceptibility to the disease of SLE, and also to detect the expression level of Peli-1 mRNA and the susceptibility to SLE and SLE. The correlation between the same phenotype, the relationship between the Peli-1 gene and the incidence of SLE from the gene transcription and expression level. The correlation between the single nucleotide polymorphisms of the peli-1 gene and systemic lupus erythematosus (SLE) in part 1 compares the difference between the SLE patients and the difference of the peli-1rs329498, rs10496105 allele and gene frequency in the normal control. The association with the genetic susceptibility of SLE. Combined with clinical data, the relationship between peli-1rs329498, rs10496105 and its main clinical features and its laboratory indicators was analyzed. Methods a case control study was used to select 738 SLE patients and 827 healthy controls. The fluidigm192.24 gene typing technique was used to detect rs329498 in the peli-1 gene. Rs10496105 two SNP loci, analysis of its genotype, allele frequency and so on. Combined with clinical data, the relationship between gene frequency distribution and clinical manifestations was further analyzed. Results (1) the association between the polymorphism of peli-1 gene rs329498 locus and SLE, the genotype frequencies of AA, AC and CC in SLE patients were 40.8%, 44.6%, and 14.6%, respectively. There was no statistically significant difference between the groups of 36.5%, 45.5%, 18% and two groups (x 2=4.63, p=0.099). The allele comparison showed that the two groups were statistically significant (x 2=4.80, p=0.028), relatively large allele A, and small allele C would reduce the risk of SLE incidence (or= 0.851,95%ci:0.737-0.983). However, there was no statistical correlation between the genetic dominance and the recessive model. Compared with the non ln nephritis group, the A allele frequency of LN patients was higher, and the difference was statistically significant (x 2=8.18, p=0.017). The or value of LN incidence risk was 0.681 (95%ci:0.528-0.880) compared with the A allele, and the difference in the genotype frequency (cc+acversusaa) distribution of the two groups was statistically significant. .632,95%ci:0.451-0.884, p=0.007). The correlation analysis of clinical features showed that the rs329498 locus of the peli-1 gene might be related to the zygomatic erythema, and the frequency distribution of the genotypes was statistically different (x 2=6.63, p=0.036). (2) the association of the peli-1 gene rs10496105 loci with SLE was associated with the rs10496105 genotype frequency GG, Ag, in the case component. No 70%, 26.2% and 3.8%, 69.4% in the control group, 27.2% and 3.4%., and no statistical difference between the control group and the control group (x 2=0.37, p=0.832). The genetic model analysis showed that no statistical difference was found between the dominant and the recessive model cases and the control groups (P0.05). In addition, the LN group and the LN group There was no statistically significant difference in genotype distribution between non LN groups (x 2=3.11, P=0.201). The results of clinical data analysis showed that there was no statistically significant association between the rs10496105 locus and the main clinical features of the zygomatic erythema, discoid erythema, light sensitivity, oral ulcer, and arthritis (P0.05). The polymorphism of the Peli-1 rs329498 gene and SLE remains in the Chinese Han population. In addition, the rs329498 locus was also associated with lupus nephritis and zygomatic erythema. No statistical association was found between the rs10496105 gene polymorphisms and SLE. Second the correlation between the Peli-1 m RNA level and systemic lupus erythematosus was compared with those in the SLE group and the normal control group, and the wolves in SLE patients. The expression level of Peil-1 gene m RNA in Peripheral blood mononuclear cells (PBMCs) in the group of ulcer nephritis and non wolf nephritis and in the active group and non active group (mononuclear cells, PBMCs), and the relationship between the expression level and clinical characteristics, the laboratory index and the degree of disease activity. Methods a total of 31 cases of SLE patients and 30 healthy pairs were collected. Trizol method was used to extract the RNA. real-time quantitative polymerase chain reaction (Real time quantitative polymerase chain reaction, RT-PCR) to detect the Peli-1 gene expression level in PBMCs, using the rank sum test to compare the differences between each group. The relationship between the level and the Systemic lupus erythematosus disease activity index, SLEDAI. All data were statistically analyzed with SPSS 17. Results (1) the level of Peli-1 m RNA in PBMCs was compared: there was no statistical difference between the 31 cases and the 30 controls. 404, P=0.686). Compared the SLE patients into the nephritis group (11 cases) and the non nephritis group (20 cases), the results showed that there was no statistical difference between the two groups of PBMCs (Z=-0.619, P=0.536). Moreover, the Peli-1 m RNA level between the active group (19 cases) and the non active group (12 cases) had no statistical difference (Z=-0.973, 2). The correlation analysis of Peli-1 m RNA level and its clinical characteristics: the Mann-Whitney rank sum test results showed that there was no statistical significance (P0.05) between the Peli-1 m RNA levels between each group. (3) the correlation between Peli-1 m level and laboratory index in PBMCs of SLE patients: the difference between the positive group and the negative group was poor. There was no statistical significance (all P0.05). (4) the level of Peli-1 m RNA expression in PBMCs of SLE patients and SLEDAI correlation analysis: Spearman grade correlation analysis showed that there was no significant correlation between Peli-1 m in SLE patients' PBMCs and the health control group. There was no statistical difference between the Peli-1 gene m RNA expression level between the LN and the non LN groups in the dynamic group and the non active group. In addition, there was no statistical difference between the major clinical manifestations and the major laboratory features, and there was no statistically significant difference between the.SLEDAI score and the RNA expression level of the Peli-1 gene m.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.241
本文编号：2127209
[Abstract]:Research background systemic lupus erythematosus (Systemic lupus erythematosus, SLE) is a kind of chronic autoimmune disease in women. Its clinical manifestation is more complex, the course of disease is prolonged, and many systems and organs are often involved. In recent years, a lot of studies have been made to explore the etiology and pathogenesis of SLE, but the exact mechanism of it has been studied. It is not yet fully understood. It is generally considered to be associated with a variety of factors such as genetic, environmental and endocrine factors that interact and interact to form a complex network of etiological factors that cause abnormal immune responses, such as the release of inflammatory factors, and the production of.SLE by autoantibodies is initially considered to be an autoimmune disease mediated by B cells. Disease. With the development of research, T cells have been found to occur in SLE and also play a vital role in the development of.1999. A highly conserved E3 ubiquitin ligase protein, named Pellino (abbreviated Peli) protein, was found in the fruit fly for the first time. The ubiquitin ligase protein family consists of Peli-1, Peli-2, Peli-3 three. The most notable functional feature of the subtype is the multiple role in innate immunity. The latest research has found that Peli-1 plays a regulatory role in a series of cellular signaling mediated by TLR/IL-1R,.Peli-1 also participates in the activation of NF- kappa B, which is mediated by TRIF dependence. In addition, Chang has found that Peli-1 is one of the maintenance of T cell tolerance. A key regulatory factor, whose functional abnormalities may lead to T cell activation, may play an important role in the pathogenesis of autoimmune diseases such as SLE. Based on the above, this subject is the first to explore the association between the Peli-1 gene polymorphism and the susceptibility to the disease of SLE, and also to detect the expression level of Peli-1 mRNA and the susceptibility to SLE and SLE. The correlation between the same phenotype, the relationship between the Peli-1 gene and the incidence of SLE from the gene transcription and expression level. The correlation between the single nucleotide polymorphisms of the peli-1 gene and systemic lupus erythematosus (SLE) in part 1 compares the difference between the SLE patients and the difference of the peli-1rs329498, rs10496105 allele and gene frequency in the normal control. The association with the genetic susceptibility of SLE. Combined with clinical data, the relationship between peli-1rs329498, rs10496105 and its main clinical features and its laboratory indicators was analyzed. Methods a case control study was used to select 738 SLE patients and 827 healthy controls. The fluidigm192.24 gene typing technique was used to detect rs329498 in the peli-1 gene. Rs10496105 two SNP loci, analysis of its genotype, allele frequency and so on. Combined with clinical data, the relationship between gene frequency distribution and clinical manifestations was further analyzed. Results (1) the association between the polymorphism of peli-1 gene rs329498 locus and SLE, the genotype frequencies of AA, AC and CC in SLE patients were 40.8%, 44.6%, and 14.6%, respectively. There was no statistically significant difference between the groups of 36.5%, 45.5%, 18% and two groups (x 2=4.63, p=0.099). The allele comparison showed that the two groups were statistically significant (x 2=4.80, p=0.028), relatively large allele A, and small allele C would reduce the risk of SLE incidence (or= 0.851,95%ci:0.737-0.983). However, there was no statistical correlation between the genetic dominance and the recessive model. Compared with the non ln nephritis group, the A allele frequency of LN patients was higher, and the difference was statistically significant (x 2=8.18, p=0.017). The or value of LN incidence risk was 0.681 (95%ci:0.528-0.880) compared with the A allele, and the difference in the genotype frequency (cc+acversusaa) distribution of the two groups was statistically significant. .632,95%ci:0.451-0.884, p=0.007). The correlation analysis of clinical features showed that the rs329498 locus of the peli-1 gene might be related to the zygomatic erythema, and the frequency distribution of the genotypes was statistically different (x 2=6.63, p=0.036). (2) the association of the peli-1 gene rs10496105 loci with SLE was associated with the rs10496105 genotype frequency GG, Ag, in the case component. No 70%, 26.2% and 3.8%, 69.4% in the control group, 27.2% and 3.4%., and no statistical difference between the control group and the control group (x 2=0.37, p=0.832). The genetic model analysis showed that no statistical difference was found between the dominant and the recessive model cases and the control groups (P0.05). In addition, the LN group and the LN group There was no statistically significant difference in genotype distribution between non LN groups (x 2=3.11, P=0.201). The results of clinical data analysis showed that there was no statistically significant association between the rs10496105 locus and the main clinical features of the zygomatic erythema, discoid erythema, light sensitivity, oral ulcer, and arthritis (P0.05). The polymorphism of the Peli-1 rs329498 gene and SLE remains in the Chinese Han population. In addition, the rs329498 locus was also associated with lupus nephritis and zygomatic erythema. No statistical association was found between the rs10496105 gene polymorphisms and SLE. Second the correlation between the Peli-1 m RNA level and systemic lupus erythematosus was compared with those in the SLE group and the normal control group, and the wolves in SLE patients. The expression level of Peil-1 gene m RNA in Peripheral blood mononuclear cells (PBMCs) in the group of ulcer nephritis and non wolf nephritis and in the active group and non active group (mononuclear cells, PBMCs), and the relationship between the expression level and clinical characteristics, the laboratory index and the degree of disease activity. Methods a total of 31 cases of SLE patients and 30 healthy pairs were collected. Trizol method was used to extract the RNA. real-time quantitative polymerase chain reaction (Real time quantitative polymerase chain reaction, RT-PCR) to detect the Peli-1 gene expression level in PBMCs, using the rank sum test to compare the differences between each group. The relationship between the level and the Systemic lupus erythematosus disease activity index, SLEDAI. All data were statistically analyzed with SPSS 17. Results (1) the level of Peli-1 m RNA in PBMCs was compared: there was no statistical difference between the 31 cases and the 30 controls. 404, P=0.686). Compared the SLE patients into the nephritis group (11 cases) and the non nephritis group (20 cases), the results showed that there was no statistical difference between the two groups of PBMCs (Z=-0.619, P=0.536). Moreover, the Peli-1 m RNA level between the active group (19 cases) and the non active group (12 cases) had no statistical difference (Z=-0.973, 2). The correlation analysis of Peli-1 m RNA level and its clinical characteristics: the Mann-Whitney rank sum test results showed that there was no statistical significance (P0.05) between the Peli-1 m RNA levels between each group. (3) the correlation between Peli-1 m level and laboratory index in PBMCs of SLE patients: the difference between the positive group and the negative group was poor. There was no statistical significance (all P0.05). (4) the level of Peli-1 m RNA expression in PBMCs of SLE patients and SLEDAI correlation analysis: Spearman grade correlation analysis showed that there was no significant correlation between Peli-1 m in SLE patients' PBMCs and the health control group. There was no statistical difference between the Peli-1 gene m RNA expression level between the LN and the non LN groups in the dynamic group and the non active group. In addition, there was no statistical difference between the major clinical manifestations and the major laboratory features, and there was no statistically significant difference between the.SLEDAI score and the RNA expression level of the Peli-1 gene m.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.241
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