瘦素和瘦素受体基因单核苷酸多态性与系统性红斑狼疮遗传易感性的关联研究

发布时间：2018-06-03 10:56

本文选题：系统性红斑狼疮 + 瘦素基因　；参考：《安徽医科大学》2017年硕士论文

【摘要】：研究背景系统性红斑狼疮(Systemic lupus erythematosus,SLE)是一种以产生大量自身抗体、累及多系统脏器、出现免疫复合物沉积为主要特征的结缔组织疾病,同时也是一种系统性的自身免疫性疾病。该疾病不仅会影响血管壁、皮肤等组织,同时还会造成关节、肾脏、血液系统及神经系统的改变。目前认为在不同地区和不同种族间SLE的发病率存在一定的差异,且男女性别间SLE发病率也存在较大差异。SLE患者中90%以上为女性,发病年龄多集中在25~45岁,目前我国女性SLE发病率约为110/10万,严重地影响了育龄期女性的身心健康。SLE的病因和发病机制至今尚未明确,可能是受遗传因素和环境因素的双重影响。瘦素是一种由瘦素基因(leptin gene,LEP)编码产生的蛋白质,参与体内多种免疫炎症过程。瘦素可以通过与瘦素受体结合形成二聚体,激活JAK2/SATA3和mTOR信号通路从而影响免疫系统功能;瘦素也可以促进T细胞增殖,抑制T细胞凋亡,诱导白细胞介素1(interleukin 1,IL-1)、IL-6、肿瘤坏死因子α(tumor necrosis factorα,TNF-α)等多种细胞因子的表达,同时促进辅助T细胞向Th1细胞转变,从而参与免疫调节过程。近期研究认为瘦素基因、瘦素受体基因(leptin receptor gene,LEPR)多态性及瘦素的表达水平与SLE、类风湿关节炎(rheumatoid arthritis,RA)等多种自身免疫性疾病的发生、发展存在关联。目前,关于SLE患者体内瘦素表达水平变化与疾病之间的关联研究结果尚有争议,且在中国汉族人群中开展的瘦素和瘦素受体基因多态性与SLE遗传易感性的关联研究非常有限。目的探讨LEP和LEPR基因单核苷酸多态性(single nucleotide polymorphism,SNP)与中国汉族人群SLE患者遗传易感性间的关联,分析LEP、LEPR基因相关snp与sle患者主要临床表现之间的关联,并比较lep、lepr基因相关snp与sle患者血清瘦素表达水平的联系。方法本课题采用病例对照研究方法,选取sle患者633例,来源于安徽医科大学附属省立医院、安徽医科大学第一附属医院和安庆市立医院三家医院风湿免疫科的门诊或住院患者,病例诊断依据美国风湿病学会1997年修订的sle分类标准。选取健康对照559例,来源于安徽医科大学第一附属医院健康体检中心健康体检者。采用多重高温连接酶检测反应技术(improvedmultipleligasedetectionreaction,imldr)进行基因分型检测,对lep基因4个位点(rs11761556,rs12706832,rs2071045,rs2167270)以及lepr基因9个位点(rs10749754,rs1137100,rs1137101,rs13306519,rs1805094,rs1805096,rs3790434,rs3806318,rs7518632)的单核苷酸多态性进行分析。采用酶联免疫吸附试验(enzymelinkedimmunosorbentassay,elisa)测定sle患者血清瘦素表达水平。结果瘦素及瘦素受体基因的13个多态性位点的基因型频率及等位基因频率在sle患者与健康对照之间的差异均无统计学意义(均有p0.05),调整性别和年龄后发现leprrs1137100位点基因型频率分布在两组间的差异存在统计学意义(ggvs.aa:χ2=3.904,p=0.048,or=1.948,95%ci:1.005-3.776)。同时,进一步分析了显性模型、隐性模型和相加模型等遗传模型在病例组与对照组之间的分布,两者之间的分布差异也均无统计学意义(均有p0.05)。调整性别和年龄后发现leprrs1137100位点的隐性模型及相加模型在两组间的差异分布存在统计学意义(ga+ggvs.aa:χ2=4.365,p=0.037,or=0.496,95%ci:0.257-0.958;ggvs.aa:χ2=3.871,p=0.049,or=0.514,95%ci:0.265-0.997)。对基因多态性与临床表现之间的关联分析发现,lep基因rs2071045位点在伴有心包炎的sle患者中,其tt基因型频率及t等位基因频率高于未伴有心包炎的sle患者(χ2=8.811,p=0.012;χ2=6.432,p=0.011)。rs11761556位点等位基因频率与sle患者是否伴随颧部红斑之间存在统计学关联(χ2=4.067,p=0.044)。lepr基因rs3806318位点基因型及等位基因频率分布与sle患者是否伴有光敏感之间具有统计学关联,sle同时伴有光敏感者其gg和ga基因型频率以及G等位基因频率高于SLE未伴有光敏感者(χ2=8.693,P=0.013;χ2=6.948,P=0.008)。同时,该位点基因型频率还与SLE患者中关节炎的发生存在统计学关联(χ2=6.204,P=0.045)。另外,在SLE合并光敏感的患者中,rs1137100位点AA和GA基因型频率及A等位基因频率高于未合并光敏感的患者(χ2=6.272,P=0.043;χ2=5.609,P=0.018)。单倍型分析发现,LEPR基因的9个位点构建的单倍型中,SLE病例组ACGCAGCAA单倍型频率低于对照组(χ2=9.038,P=0.003,OR=0.745,95%CI:0.615-0.903);而ATGCAGCAA单倍型结构在SLE病例组中的频率高于健康对照组(χ2=4.327,P=0.038,OR=1.390,95%CI:1.018-1.897)。SLE患者的瘦素血清表达水平与13个位点单核苷酸多态性之间均无统计学关联(均有P0.05)。结论瘦素和瘦素受体基因的13个位点SNP与SLE遗传易感性无统计学关联,但调整性别、年龄后发现LEPR rs1137100位点基因型频率在两组中分布存在差异,且SLE患者不同瘦素和瘦素受体基因型下的的血清瘦素表达水平差异无统计学意义,但部分位点SNP可能与该疾病的某些临床表型有关。
[Abstract]:Background systemic lupus erythematosus (Systemic lupus erythematosus) is a kind of connective tissue disease characterized by the production of a large number of autoantibodies, involving multiple system organs and the deposition of immune complex. It is also a systemic autoimmune disease. The disease not only affects the blood vessel wall, skin and other tissues, but also the disease. There are also changes in the joint, kidney, blood system and nervous system. At present, there is a certain difference in the incidence of SLE between different regions and different races, and the incidence of SLE among men and women is also significantly different. More than 90% of the.SLE patients are women, the age of onset is at the age of 25~45, and the incidence of SLE in Chinese women is about 11. 0/10 million, seriously affecting the physical and mental health of women of childbearing age, the etiology and pathogenesis of.SLE is not yet clear. It may be affected by both genetic and environmental factors. Leptin is a protein encoded by the leptin gene (LEP) and participates in a variety of immune inflammatory processes in the body. Leptin can be affected by leptin. Body combination forms two polymer and activates JAK2/SATA3 and mTOR signaling pathways that affect the function of the immune system, and leptin can also promote the proliferation of T cells, inhibit the apoptosis of T cells, induce the expression of interleukin 1 (interleukin 1, IL-1), IL-6, tumor necrosis factor alpha (tumor necrosis factor A, TNF- alpha) and so on, and promote the auxiliary T thin. In recent studies, the expression of leptin gene, leptin receptor gene (leptin receptor gene, LEPR) and the expression level of leptin are associated with the development of many autoimmune diseases such as SLE, rheumatoid arthritis (rheumatoid arthritis, RA) and other autoimmune diseases. There is a relationship between the development of the leptin gene and leptin receptor gene (rheumatoid arthritis, RA) and other autoimmune diseases. Currently, it is related to the body of SLE patients. The correlation between leptin expression level and disease is controversial, and the association of leptin and leptin receptor gene polymorphism with SLE genetic susceptibility in Chinese Han population is very limited. The purpose of this study is to explore the single nucleotide polymorphisms of LEP and LEPR genes (single nucleotide polymorphism, SNP) and Chinese Han people Association of genetic susceptibility to group SLE patients, analysis of the association between LEP, LEPR gene related SNP and the main clinical manifestations of SLE patients, and compare the relationship between LEP, LEPR gene related SNP and the level of serum leptin expression in SLE patients. Methods a case control study was used to select 633 cases of SLE patients, from the affiliated province of Medical University Of Anhui. The hospital, the First Affiliated Hospital of Medical University Of Anhui and the three hospitals of the Department of Rheumatology in the Department of Rheumatology, three hospitals and hospitalized patients, were diagnosed according to the SLE classification standard revised by the American rheumatology society in 1997. 559 healthy controls were selected from the health checkup in the health check-up center of the First Affiliated Hospital of Medical University Of Anhui. Improvedmultipleligasedetectionreaction (imldr) was used to detect the genotyping of the 4 loci of the LEP gene (rs11761556, rs12706832, rs2071045, rs2167270) and the 9 loci of the LEPR gene. The single nucleotide polymorphisms were analyzed. The serum leptin expression level in SLE patients was measured by enzymelinkedimmunosorbentassay (ELISA). The genotype frequencies and allele frequencies of 13 polymorphic loci of leptin and leptin receptor genes were not statistically significant between SLE patients and healthy controls. Meaning (all P0.05), after adjusting sex and age, it was found that the difference in the frequency distribution of leprrs1137100 loci in the two groups was statistically significant (ggvs.aa: x 2=3.904, p=0.048, or=1.948,95%ci:1.005-3.776). Meanwhile, the genetic models of dominant, recessive and additive models were further analyzed between the case group and the control group. There was no statistical significance (all P0.05) in the distribution difference between the two groups. The difference distribution between the two groups was statistically significant (ga+ggvs.aa: x 2=4.365, p=0.037, or=0.496,95%ci:0.257-0.958, ggvs.aa: Chi 2=3.871, p=0.049, or=0.514,95%ci) after adjusting the sex and age of the leprrs1137100 site. 0.265-0.997. Analysis of the association between gene polymorphism and clinical manifestations found that the frequency of the TT genotype frequency and T allele frequencies of the LEP gene rs2071045 locus in the SLE patients with pericarditis were higher than those of the SLE patients without pericarditis (x 2=8.811, p=0.012; Chi 2=6.432, p=0.011) allele frequencies with those of the patients. There was a statistical correlation between the zygomatic erythema (x 2=4.067, p=0.044).Lepr gene rs3806318 genotype and the allele frequency distribution with the light sensitivity of SLE patients. The frequencies of GG and GA genotype and G allele frequencies of SLE and G alleles were higher than those of SLE without photosensitivity ( X 2=8.693, P=0.013; X 2=6.948, P=0.008). At the same time, the genotype frequency of this loci was also associated with the occurrence of arthritis in SLE patients (x 2=6.204, P=0.045). In addition, the frequency of AA and GA genotypes and A alleles at rs1137100 loci were higher in patients with SLE with light sensitivity than those without light sensitivity. 2=5.609, P=0.018). The haplotype analysis found that in the haplotype of the 9 loci of LEPR gene, the frequency of ACGCAGCAA haplotype in SLE case group was lower than that of the control group (x 2=9.038, P=0.003, OR=0.745,95%CI:0.615-0.903), and the frequency of ATGCAGCAA haplotype in SLE case group was higher than that of the healthy control group (x 2=4.327, P=0.038, etc.) 897) there was no statistical correlation between the serum level of leptin in.SLE patients and the single nucleotide polymorphisms of the 13 loci (all P0.05). Conclusion the 13 loci of leptin and leptin receptor gene SNP were not associated with the genetic susceptibility to SLE, but the genotype frequencies of the LEPR rs1137100 loci were found to be distributed in the two groups after the adjustment of sex. There is no significant difference in the level of serum leptin expression in SLE patients with leptin and leptin receptor genotypes, but some of the site SNP may be related to some clinical phenotypes of the disease.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R593.241

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