發(fā)布時間：2018-03-03 22:14

  本文選題：視神經(jīng)脊髓炎譜系疾病　切入點：腫瘤壞死因子　出處：《河北醫(yī)科大學(xué)》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:視神經(jīng)脊髓炎譜系疾病(neuromyelitis optica spectrum disorder,NMOSD)是一種免疫介導(dǎo)的中樞神經(jīng)系統(tǒng)炎性脫髓鞘疾病,常于青壯年起病,女性多與男性,其病程呈復(fù)發(fā)-緩解或進行性加重,復(fù)發(fā)率及致殘率高,預(yù)后較差。許多患者留有感覺異常、運動障礙等明顯的后遺癥,不僅嚴(yán)重影響了患者的身心健康,還給不少家庭帶來經(jīng)濟困難。其確切發(fā)病原因及機制尚不明確,涉及遺傳、免疫、環(huán)境等因素。自身免疫性疾病常常存在基因易感性,腫瘤壞死因子(tumor necrosis factor,TNF)包括兩種細(xì)胞因子—腫瘤壞死因子α和腫瘤壞死因子β(TNFα和TNFβ),其在免疫應(yīng)答過程中具有廣泛調(diào)節(jié)免疫的作用。其基因調(diào)節(jié)區(qū)域的基因多態(tài)性可以影響自身的分泌,與自身免疫性疾病的相關(guān)關(guān)系已被廣泛研究,與NMOSD的相關(guān)性研究未見報道,本研究探討腫瘤壞死因子及其基因多態(tài)性與視神經(jīng)脊髓炎譜系疾病的相關(guān)關(guān)系。方法:本實驗通過收錄2015年9月1日—2016年8月30日于河北醫(yī)科大學(xué)第二醫(yī)院神經(jīng)內(nèi)科門診及住院的符合2015年視神經(jīng)脊髓炎譜系疾病診斷標(biāo)準(zhǔn)國際共識的患者50例,抽取空腹靜脈血直接保存用于檢測基因多態(tài)性,于患者激素治療前及經(jīng)治療好轉(zhuǎn)后緩解期分別抽取空腹肘正中靜脈血離心,留取血清用于測定TNFα和TNFβ的含量,同時收集年齡、性別、EDSS評分等臨床資料。收集同期年齡、性別等相匹配的健康體檢者作為對照組留取血液標(biāo)本。用連接酶檢測反應(yīng)分型方法對TNFα啟動子區(qū)-308位點(rs1800629)和TNFβ內(nèi)含子區(qū)+252位點(rs909253)基因分型;用ELISA方法測定NMOSD急性期、緩解期及健康對照組TNFα和TNFβ的含量;應(yīng)用SPSS 20.0統(tǒng)計軟件對實驗數(shù)據(jù)進行統(tǒng)計學(xué)分析。結(jié)果:1一般臨床資料本研究入選NMOSD組患者50例,健康對照組60例,兩組在年齡、性別構(gòu)成比上無統(tǒng)計學(xué)差異。2基因型分布及等位基因頻率比較2.1各組基因型分布的哈迪-溫伯格平衡定律檢測(Hardy-Weinberg equilibrium)2.1.1 NMOSD組TNFα基因-308位點(rs1800629)各基因型實際數(shù)與預(yù)期數(shù)比較,沒有顯著差異(P0.05);TNFβ基因+252位點(rs909253)各基因型實際數(shù)與預(yù)期數(shù)比較,沒有顯著差異(P0.05),說明NMOSD組樣本具有群體代表性。2.1.2正常對照組TNFα-308位點(rs1800629)各基因型實際數(shù)與預(yù)期數(shù)比較,沒有顯著差異(P0.05);TNFβ+252位點(rs909253)各基因型實際數(shù)與預(yù)期數(shù)相比較,沒有顯著差異(P0.05),說明正常對照組樣本具有群體代表性。2.2各組間基因型分布和等位基因頻率的比較2.2.1 NMOSD患者與正常對照組TNFα基因型分布及等位基因頻率比較差異無統(tǒng)計學(xué)意義。2.2.2 NMOSD患者與正常對照組TNFβ基因型分布及等位基因頻率比較差異無統(tǒng)計學(xué)意義。TNFβ基因型CC與CT+TT比較基因型分布差異無統(tǒng)計學(xué)意義。3各組血清中TNFα水平和TNFβ水平比較3.1各組間血清TNFα水平的比較NMOSD急性期患者血清TNFα38例,均值水平為:16.97(25.75)pg/ml;緩解期39例均值水平為:9.39(7.48)pg/ml;健康對照組60例均值水平為12.89(12.08)pg/ml;NMOSD組急性期TNFα水平高于緩解期(P0.05);NMOSD組急性期TNFα水平高于正常對照組(P0.05);NMOSD組緩解期TNFα水平低于正常對照組(P0.05)。3.2各組間血清TNFβ水平的比較NMOSD組急性期血清TNFβ38例,均值水平為:75.88(39.60)pg/ml;緩解期39例均值水平為:56.82(34.48)pg/ml;健康對照組60例均值水平為67.77(35.48)pg/ml;NMOSD組急性期TNFβ水平高于緩解期(P0.05);NMOSD組急性期TNFβ均值水平高于健康對照組,但差異無統(tǒng)計學(xué)意義(P0.05)。緩解期NMOSD組TNFβ水平低于正常對照組(P0.05)。TNFα不同基因型急性期血清TNFα水平差異無統(tǒng)計學(xué)意義;TNFβ不同基因型急性期血清TNFβ水平差異無統(tǒng)計學(xué)意義。5基因型與臨床資料相關(guān)關(guān)系5.1 TNFα各基因型NMOSD患者發(fā)病年齡,發(fā)病次數(shù)、急性期EDSS評分,EDSS評分差值(急性期-緩解期)進行組間比較差異均無統(tǒng)計學(xué)意義;TNFα各基因型AQP4-Ig G陽性率的比較差異無統(tǒng)計學(xué)意義。5.2 TNFβ各基因型NMOSD患者發(fā)病年齡,發(fā)病次數(shù)、急性期EDSS評分,EDSS評分差值(急性期-緩解期)進行組間比較差異均無統(tǒng)計學(xué)意義;TNFβ各基因型AQP4-Ig G陽性率的比較差異無統(tǒng)計學(xué)意義。6不同時期TNFα和TNFβ血清水平與同時期EDSS評分及TNFα和TNFβ血清水平之間關(guān)系急性期、緩解期EDSS評分分別與同時期TNFα、TNFβ血清水平之間均無顯著相關(guān)性(P0.05);TNFα急性期、緩解期血清水平與同時期TNFβ急性期、緩解期血清水平均無顯著相關(guān)性(P0.05)。4 TNF不同基因型對急性期血清TNF水平的影響結(jié)論:1本實驗研究顯示TNFα基因(rs1800629)和TNFβ基因(rs909253)的多態(tài)性與NMOSD患者的易感性不相關(guān)。2 NMOSD組血清TNFα水平急性期高于緩解期和健康對照組,緩解期TNFα水平低于健康對照組,TNFα可能與NMOSD疾病發(fā)病相關(guān),在其發(fā)病機制中起作用。3 NMOSD患者血清TNFβ水平在急性期高于緩解期;急性期均值水平高于健康對照組,但差異無統(tǒng)計學(xué)意義;緩解期TNFβ水平均低于正常對照組,TNFβ可能與NMOSD疾病發(fā)病相關(guān),在其發(fā)病機制中起作用。4 TNFα(rs1800629)基因多態(tài)性對急性期血清TNFα水平無影響。TNFβ(rs909253)基因多態(tài)性對急性期血清TNFβ水平無影響。5 TNFα和TNFβ基因多態(tài)性與NMOSD患者發(fā)病年齡,發(fā)病次數(shù)、急性期EDSS評分,EDSS評分差值(急性期-緩解期)、AQP4-Ig G陽性率均無顯著相關(guān)性。6急性期、緩解期EDSS評分分別與同時期TNFα、TNFβ血清水平之間均無相關(guān)性;TNFα急性期、緩解期血清水平與同時期TNFβ急性期、緩解期血清水平均無相關(guān)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.

【學(xué)位授予單位】：河北醫(yī)科大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R744.52

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本文編號：1562918