肿瘤坏死因子及其基因多态性与视神经脊髓炎谱系疾病的相关性研究

发布时间：2018-03-03 22:14

本文选题：视神经脊髓炎谱系疾病　切入点：肿瘤坏死因子　出处：《河北医科大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:视神经脊髓炎谱系疾病(neuromyelitis optica spectrum disorder,NMOSD)是一种免疫介导的中枢神经系统炎性脱髓鞘疾病,常于青壮年起病,女性多与男性,其病程呈复发-缓解或进行性加重,复发率及致残率高,预后较差。许多患者留有感觉异常、运动障碍等明显的后遗症,不仅严重影响了患者的身心健康,还给不少家庭带来经济困难。其确切发病原因及机制尚不明确,涉及遗传、免疫、环境等因素。自身免疫性疾病常常存在基因易感性,肿瘤坏死因子(tumor necrosis factor,TNF)包括两种细胞因子—肿瘤坏死因子α和肿瘤坏死因子β(TNFα和TNFβ),其在免疫应答过程中具有广泛调节免疫的作用。其基因调节区域的基因多态性可以影响自身的分泌,与自身免疫性疾病的相关关系已被广泛研究,与NMOSD的相关性研究未见报道,本研究探讨肿瘤坏死因子及其基因多态性与视神经脊髓炎谱系疾病的相关关系。方法:本实验通过收录2015年9月1日—2016年8月30日于河北医科大学第二医院神经内科门诊及住院的符合2015年视神经脊髓炎谱系疾病诊断标准国际共识的患者50例,抽取空腹静脉血直接保存用于检测基因多态性,于患者激素治疗前及经治疗好转后缓解期分别抽取空腹肘正中静脉血离心,留取血清用于测定TNFα和TNFβ的含量,同时收集年龄、性别、EDSS评分等临床资料。收集同期年龄、性别等相匹配的健康体检者作为对照组留取血液标本。用连接酶检测反应分型方法对TNFα启动子区-308位点(rs1800629)和TNFβ内含子区+252位点(rs909253)基因分型;用ELISA方法测定NMOSD急性期、缓解期及健康对照组TNFα和TNFβ的含量;应用SPSS 20.0统计软件对实验数据进行统计学分析。结果:1一般临床资料本研究入选NMOSD组患者50例,健康对照组60例,两组在年龄、性别构成比上无统计学差异。2基因型分布及等位基因频率比较2.1各组基因型分布的哈迪-温伯格平衡定律检测(Hardy-Weinberg equilibrium)2.1.1 NMOSD组TNFα基因-308位点(rs1800629)各基因型实际数与预期数比较,没有显著差异(P0.05);TNFβ基因+252位点(rs909253)各基因型实际数与预期数比较,没有显著差异(P0.05),说明NMOSD组样本具有群体代表性。2.1.2正常对照组TNFα-308位点(rs1800629)各基因型实际数与预期数比较,没有显著差异(P0.05);TNFβ+252位点(rs909253)各基因型实际数与预期数相比较,没有显著差异(P0.05),说明正常对照组样本具有群体代表性。2.2各组间基因型分布和等位基因频率的比较2.2.1 NMOSD患者与正常对照组TNFα基因型分布及等位基因频率比较差异无统计学意义。2.2.2 NMOSD患者与正常对照组TNFβ基因型分布及等位基因频率比较差异无统计学意义。TNFβ基因型CC与CT+TT比较基因型分布差异无统计学意义。3各组血清中TNFα水平和TNFβ水平比较3.1各组间血清TNFα水平的比较NMOSD急性期患者血清TNFα38例,均值水平为:16.97(25.75)pg/ml;缓解期39例均值水平为:9.39(7.48)pg/ml;健康对照组60例均值水平为12.89(12.08)pg/ml;NMOSD组急性期TNFα水平高于缓解期(P0.05);NMOSD组急性期TNFα水平高于正常对照组(P0.05);NMOSD组缓解期TNFα水平低于正常对照组(P0.05)。3.2各组间血清TNFβ水平的比较NMOSD组急性期血清TNFβ38例,均值水平为:75.88(39.60)pg/ml;缓解期39例均值水平为:56.82(34.48)pg/ml;健康对照组60例均值水平为67.77(35.48)pg/ml;NMOSD组急性期TNFβ水平高于缓解期(P0.05);NMOSD组急性期TNFβ均值水平高于健康对照组,但差异无统计学意义(P0.05)。缓解期NMOSD组TNFβ水平低于正常对照组(P0.05)。TNFα不同基因型急性期血清TNFα水平差异无统计学意义;TNFβ不同基因型急性期血清TNFβ水平差异无统计学意义。5基因型与临床资料相关关系5.1 TNFα各基因型NMOSD患者发病年龄,发病次数、急性期EDSS评分,EDSS评分差值(急性期-缓解期)进行组间比较差异均无统计学意义;TNFα各基因型AQP4-Ig G阳性率的比较差异无统计学意义。5.2 TNFβ各基因型NMOSD患者发病年龄,发病次数、急性期EDSS评分,EDSS评分差值(急性期-缓解期)进行组间比较差异均无统计学意义;TNFβ各基因型AQP4-Ig G阳性率的比较差异无统计学意义。6不同时期TNFα和TNFβ血清水平与同时期EDSS评分及TNFα和TNFβ血清水平之间关系急性期、缓解期EDSS评分分别与同时期TNFα、TNFβ血清水平之间均无显著相关性(P0.05);TNFα急性期、缓解期血清水平与同时期TNFβ急性期、缓解期血清水平均无显著相关性(P0.05)。4 TNF不同基因型对急性期血清TNF水平的影响结论:1本实验研究显示TNFα基因(rs1800629)和TNFβ基因(rs909253)的多态性与NMOSD患者的易感性不相关。2 NMOSD组血清TNFα水平急性期高于缓解期和健康对照组,缓解期TNFα水平低于健康对照组,TNFα可能与NMOSD疾病发病相关,在其发病机制中起作用。3 NMOSD患者血清TNFβ水平在急性期高于缓解期;急性期均值水平高于健康对照组,但差异无统计学意义;缓解期TNFβ水平均低于正常对照组,TNFβ可能与NMOSD疾病发病相关,在其发病机制中起作用。4 TNFα(rs1800629)基因多态性对急性期血清TNFα水平无影响。TNFβ(rs909253)基因多态性对急性期血清TNFβ水平无影响。5 TNFα和TNFβ基因多态性与NMOSD患者发病年龄,发病次数、急性期EDSS评分,EDSS评分差值(急性期-缓解期)、AQP4-Ig G阳性率均无显著相关性。6急性期、缓解期EDSS评分分别与同时期TNFα、TNFβ血清水平之间均无相关性;TNFα急性期、缓解期血清水平与同时期TNFβ急性期、缓解期血清水平均无相关性。
[Abstract]:Objective: neuromyelitis optica spectrum disease (neuromyelitis optica spectrum disorder, NMOSD) is an immune-mediated demyelinating disease, often in young women and men, the onset, the disease was aggravated or relapse remission, relapse rate and high disability rate, poor prognosis. Patients have abnormal sensation, movement disorders and other obvious sequelae, not only seriously affect the patient's physical and mental health, but also brings economic difficulties family. Its pathogenesis and mechanism is not clear, involving genetic, immune, environment and other factors. Autoimmune diseases often have a genetic susceptibility, tumor necrosis factor (tumor necrosis, factor TNF), including two kinds of cytokines, tumor necrosis factor alpha and tumor necrosis factor beta (TNF alpha and TNF beta), which has a broad immunoregulatory role in immune response. The gene transfer Gene polymorphism can affect the regional section secretion, associated with autoimmune diseases has been extensively studied, and the correlation of NMOSD has not been reported. This study was to investigate the relationship between tumor necrosis factor and the gene polymorphism and neuromyelitis optica spectrum disorders. Methods: This study included 50 patients by September 1, 2015 August 30, 2016 in the second hospital of Hebei Medical University Department of neurology outpatient and inpatient meet 2015 neuromyelitisoptica spectrum diagnosis standard international consensus, used to detect the gene polymorphism directly saved fasting venous blood was drawn from patients with hormone therapy before and after treatment. After the remission stage were collected fasting venous blood Zhou Zhengzhong centrifugal, serum for content determination of TNF alpha and TNF beta, while collecting the clinical data of age, gender, EDSS score and so on. At the same period were age, gender etc. Matched healthy subjects as control group. Blood sample was collected for classification with ligase detection reaction promoter of TNF alpha locus -308 (rs1800629) and TNF beta intron +252 loci (rs909253) genotype; Determination of acute NMOSD with ELISA method, remission and healthy control group was TNF alpha and beta TNF; application of SPSS 20 statistical software for statistical analysis of experimental data. Results: 1 clinical data in this study included 50 NMOSD patients, 60 healthy subjects served as control group, two groups in age, than no statistical difference between.2 genotype and allele frequency distribution were compared between the 2.1 groups of genotype distribution the Hardy Weinberg equilibrium detection (Hardy-Weinberg equilibrium) 2.1.1 sex group NMOSD TNF alpha -308 gene (rs1800629) genotypes compared actual and expected number, there was no significant difference (P0.05); TNF gene locus +252 (rs909253 Comparison of various genotypes) actual and expected number, there was no significant difference (P0.05), NMOSD group of samples with a representative group.2.1.2 normal control group TNF alpha -308 loci (rs1800629) comparing the genotypes of actual and expected number, there was no significant difference (P0.05); TNF beta +252 point (rs909253) of each gene the actual number and the expected number of comparison, no significant difference (P0.05), normal control group samples with a representative group of.2.2 groups between genotype distribution and allele frequencies of NMOSD between 2.2.1 patients and normal controls, and the distribution of allele frequency differences between the group of TNF alpha gene type was not statistically significant.2.2.2 in patients with NMOSD the normal distribution and allele frequency differences between group TNF beta gene type was not statistically significant.TNF beta CC genotype compared with CT+TT genotype had no significant difference in the serum.3 level of TNF alpha and TNF beta level than The level of serum TNF 3.1 between groups were compared in patients with acute NMOSD serum TNF in 38 cases, the average level is 16.97 (25.75): pg/ml; 39 cases in remission period average: 9.39 (7.48) pg/ml; 60 cases of healthy control groups mean level of 12.89 (12.08) pg/ml; TNF levels in group NMOSD acute higher than remission (P0.05); the TNF alpha level of NMOSD group in acute period is higher than the normal control group (P0.05); NMOSD group of remission TNF a level lower than the normal control group (P0.05) level of serum beta TNF.3.2 groups NMOSD group in acute phase serum TNF beta 38 cases, the average level is 75.88 (39.60) pg/ml; 39 cases in remission period average: 56.82 (34.48) pg/ml; 60 cases of healthy control groups mean level of 67.77 (35.48) pg/ml; TNF beta level NMOSD group in acute stage than in remission (P0.05); the average level of TNF beta in acute stage of NMOSD higher than the healthy control group, but the difference was not statistically significant (P0.05). The remission group NMOSD beta TNF The level is lower than the normal control group (P0.05) there was no significant difference in serum levels of TNF alpha.TNF alpha genotypes in acute stage; differences in serum TNF TNF beta genotypes in acute phase was not statistically significant.5 genotype and clinical data of 5.1 patients with primary disease between age TNF alpha genotypes NMOSD incidence, acute period EDSS score, EDSS score (acute - remission) were compared between the two groups had no significant difference; the difference of positive rate of AQP4-Ig G TNF alpha genotypes had no statistical significance with age of onset,.5.2 TNF beta genotype NMOSD incidence, acute EDSS score, EDSS score (acute period remission) were compared between the two groups had no significant difference; no statistical significance of.6 in different stages of TNF alpha and TNF beta serum level and at the same time, EDSS score and TNF alpha and TNF difference between the positive rate of AQP4-Ig G TNF beta gene types The relationship between serum beta levels in acute phase, remission of EDSS score respectively with the same period of TNF alpha, TNF beta between serum level showed no significant correlation (P0.05); TNF alpha in acute phase, remission serum level with the same period of TNF beta acute phase, remission serum level showed no significant correlation (P0.05) influence the serum level of TNF.4 TNF of different genotypes on the acute phase of conclusion: TNF alpha gene 1 the experimental research shows that the (rs1800629) and TNF gene (rs909253) polymorphism and susceptibility to NMOSD were not related to.2 group NMOSD serum TNF levels in acute stage than in remission and healthy control group, remission TNF levels lower than the healthy the control group, the incidence of TNF and NMOSD may be relevant in the pathogenesis of disease, play a role in.3 NMOSD TNF in serum of patients with beta level is higher than the remission in acute period; the average level of acute phase than in healthy controls, but the difference was not statistically significant; remission of TNF beta The level is lower than that of normal control group, the incidence of TNF and NMOSD may be related to beta.4 disease, play a role in the pathogenesis of TNF alpha (rs1800629) serum levels of TNF alpha gene polymorphism in acute stage of.TNF had no effect on beta (rs909253).5 had no effect on TNF alpha and TNF beta gene polymorphisms in patients with NMOSD, age of onset, serum the level of TNF beta gene polymorphism on acute onset of acute phase frequency, EDSS score, EDSS score (acute period and remission period), the positive rate of AQP4-Ig G was no significant correlation between.6 in acute phase, remission of EDSS score respectively with the same period of TNF alpha, between the level of serum TNF was no correlation between beta TNF alpha in acute stage; the serum level of remission, with the same period of TNF beta acute phase, remission serum levels were not correlated.

【学位授予单位】：河北医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R744.52

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