【作者】 沙龙金；

【导师】 漆松涛；

【作者基本信息】 南方医科大学 ， 神经外科（专业学位）， 2014， 博士

【摘要】 颅内动脉瘤是由于颅内局部血管异常改变,导致颅内动脉壁产生囊性膨出,形成的脑血管瘤样突起。颅内动脉瘤80%发生于脑底动脉环前半部,是脑血管系统中具有破裂倾向的局灶性病理扩张,其破裂可致脑蛛网膜下腔出血,因而是目前最具破坏性的脑血管疾病之一。研究表明,85%的SAH是由于颅内动脉瘤破裂而引起的,动脉瘤破裂出血后24h内的病死率达到2①%,1周内的病死率达到40%,出血后1个月内的病死率高达50%,而在存活者中,1／3以上患者遗留严重的神经功能缺损。且颅内动脉瘤的发病率只为1%～8%,不及脑血栓,高血压脑出血排在颅内病变的第3位,但其高病死率、高致残率,给家庭和社会带来了沉重负担。颅内动脉瘤的发病是多因素引起的,目前研究认为,体力劳动、情绪波动、酒后、解便等均可引起动脉瘤突然破裂,而年龄、性别、吸烟、饮酒、高血压已被证实是颅内动脉瘤的危险因素。炎症指循环系统中的活体组织对损伤因子所发生的防御,血管反应是炎症过程的中心环节。炎性细胞介导的异常血管重塑,被认为是组成各种血管疾病的中的一个关键的病理环节。血流变化也可激活内皮细胞和炎性细胞释放炎性介质,血管壁剪应力的升高可以上调IL-1β的水平,使炎性细胞易于浸润血管壁,从而与活化的内皮细胞、平滑肌细胞等共同介导炎性反应。炎性细胞、抗体和补体等参与的炎性反应,诱导了多种蛋白酶表达是导致血管壁损伤并形成动脉瘤的原因,但颅内动脉瘤形成机制相关的具体信号通路尚不清楚。TNF-α是白细胞产生的关键免疫活性物质,在颅内动脉瘤中显著增加,强烈地活化血管内皮细胞。此外,颅内动脉瘤破裂出血后血液及脑脊液中IL-6也明显增高。遗传是指经过基因的传递,使后代获得类似亲代特征的过程。DNA作为遗传物质,通过转录和翻译的过程,决定了生命的基本特性,也与疾病的发生息息相关。遗传学因素在颅内动脉瘤的发病机制上具有重要意义,流行病学调查发现,部分颅内动脉瘤患者呈现家族性发病倾向,患者一级亲属发生该病的风险明显增高。芬兰东部人群家族性颅内动脉瘤患者的研究中发现,颅内动脉瘤阳性家族史患者发病风险率可增加12%。有颅内动脉瘤家族史的日本人群中发现,未破裂动脉瘤的患病率显著增高10.5%-13.5%。因此,遗传因素在颅内动脉瘤的发生中起着重要作用。SNP位点是指在基因组上单个核苷酸的变异形成的遗传标记,其数量很多,多态性丰富,是第三代遗传标志。人体许多表型差异、对药物或疾病的易感性等都可能与SNP有关。尽管大多数SNP位于基因组非编码元件内含子中,但它在RNA剪切过程中可能有重要作用。利用全基因组SNP扫描对不同种族的颅内动脉瘤家系进行研究,证实了多个染色体1p34.3-lp36.13、7q11、19q13.3及Xp22区段可能与动脉瘤的发生相关。此外,通过传统的候选基因研究法,通过对与颅内动脉瘤发生功能相关的基因进行测定,也发现了许多的易感基因,如内皮糖蛋白基因、基质金属蛋白酶基因、IR型胶原基因、otl-抗胰蛋白酶基因、弹力蛋白基因、血管紧张素转化酶基因、载脂蛋白基因等,其中有一些基因功能与血管壁的形成相关,因而也从分子功能上得到一系列的证据与颅内动脉瘤相关。一些研究结果也表明,某些炎症细胞因子也与颅内动脉瘤有正相关性,IL-1p, IL-6及TNF-α等与颅内动脉瘤的易感性相关联。尽管目前已有报道TNF-α、IL-1β及IL-6与颅内动脉瘤的易感性相关,但是它们却没有在GWAS大样本、全基因组的数据中得到验证。在意大利人群中IL-6不是颅内动脉瘤的易感基因。因此,TNF-α、IL-1β及IL-6是否是颅内动脉瘤的易感基因仍不确定,这可能是种族人群之间的遗传差异性造成的,也可能是候选基因组研究中样本数小、质控不严及统计学分析策略不严密得到的错误的相关性信息。为了进一步的证实TNF-α、IL-1β及IL-6基因多态性与中国汉族人群颅内动脉瘤的相关性,本研究通过收集168例颅内动脉瘤患者及严格匹配的184名同龄正常体检者,对TNF-α、IL-1β及IL-6基因附近的标签SNP位点进行基因分型,通过严格的质控及严谨的统计学分析,以探索TNF-α、IL-1β及IL-6基因的多态性是否与中国汉族人群颅内动脉瘤的发病相关。这一课题的实施将使得我们找到颅内动脉瘤的发病机制成为可能,为颅内动脉瘤易感基因的确定、易感人群的筛选、早期诊断和治疗提供了分子生物学基础,对不断提高疾病检出率、提高患者生存率和生存质量具有重要的意义。第一部分炎症相关基因多态性与168例颅内动脉瘤易感性的关联研究1.目的探讨颅内动脉瘤患者与正常对照者中炎症相关基因区域内SNPs位点的等位基因频率与基因型频率分布情况。探讨炎症相关基因多态性与颅内动脉瘤后SAH的严重程度、临床预后及颅内动脉瘤发病数量(单发性／多发性)之间的关联性。2.材料与方法2.1研究对象：研究组为2010～2012年间于南方医院神经外科病房收集168名颅内动脉瘤性SAH伴或不伴脑内血肿的患者。患者为发病时间<72小时,年龄>18岁成人患者,颅内动脉瘤均经临床症状及全脑数字减影血管成像和或CT血管成像确诊为颅内动脉瘤性。对照组为184名对照组受试者为本院体检科体检的健康汉族人群,所有受试者均无颅内动脉瘤病史。SNP位点为NCBI数据库中挑选出TNF-α、IL-1β及IL-6基因区域内3个代表性SNPs位点(rs1800795、rs1143627及rs1799964)。2.2研究方法：记录门诊颅内动脉瘤患者、入院治疗颅内动脉瘤患者及健康体检者一般临床资料,制表并输入数据至Excel中。用天根生化公司全血基因组DNA提取试剂盒提取外周血全基因组DNA。得到的基因组DNA片段用琼脂糖凝胶电泳鉴定是否降解,并在Nanodrop ND-2000紫外分光光度计上测量浓度。在NCBI数据库中根据SNP位点所在的位置查询相应的碱基序列设计PCR引物,并用基因组DNA扩增相应的片段。最后在生物公司中测序鉴定各个样本SNP位点的基因型。2.3统计学处理：使用SPSS统计软件的卡方检验程序进行等位基因频率和基因型频率统计,染色体TNF-α、IL-1β及IL-6基因区域标签SNP位点在病例组和对照组之间等位基因和基因型频率差异比较用Z检验。计量资料用(均数±标准差)表示,经方差齐性检验后(P>0.05)采用单因素方差分析对组间的均数进行比较。P<0.05认为差异有统计学意义。采用Logist回归模型中的“逐步模型”对主效SNP位点进行鉴定,并评估易感SNP位点与颅内动脉瘤相关危险因素之间是否存在累积效应。使用Haploview4.2软件进行中国及日本人群特定染色体区段SNP位点的哈-温平衡检验和连锁不平衡(Linkage disequilibrium, LD)分析,并绘制该区段的LD图。3.结果本研究组共收集168名颅内动脉瘤患者,其中男性74例(44.05%),女性94例(55.95%),平均年龄(51.67+18.71)岁,BMI为(23.21±2.16)。两组间一般情况,包括高血压史、高血脂史、糖尿病史、吸烟史及饮酒史无显著性差异(均P>0.05),因此具有可比性。颅内动脉瘤患者经治疗后转归情况用Glasgow预后评分(Glasgow Outcome Scale, GOS)进行评估,GOS4～5分为预后良好,GOS1～3分为预后不佳。其中预后良好患者122例(占72.62%)、预后不佳患者46例(占27.38%)。在UCSC基因库中寻找出待测基因TNF-α、IL-1β及IL-6所处的区域：分别在以上三个基因所在区域外的10kb范围内(上下游各5kb)寻找中国人群该区域的SNP位点,SNP位点选择的原则：豢最月、等位基因频率大于0.1(由于本研究样本较少,如果选择频率较小的SNP位点,组间SNP位点的基因型个数太少,无法进行统计)。2连锁不平衡原则：当SNP位点位于连锁度较高的区域内时,可以选择其中任意一个,其余SNP位点都可以被该位点所替代。3SNP位点所处的位置：内含子区域的SNP位点功能作用的发挥未知,尽量选择位于上游调控区的位点。最终我们选择位于TNF-α基因上游的rs1799964、位于IL-1β基因上游的rs1143627、位于IL-6基因上游的具有代表该区域的rs1800795作为我们的待检SNP位点。颅内动脉瘤及健康受试者样本全血DNA浓度在100ng-1μg之间,质量良好,琼脂糖凝胶电泳显示片段大小在15000kb以上,无拖尾现象,示DNA无降解。TNF-α、IL-1β及IL-6SNP位点rsl799964、rs1143627及rs1800795周围PCR扩增片段琼脂糖凝胶电泳所示,均为单一性条带,无非特异性产物扩增,根据PCR产物的序列,鉴定样本各个SNP位点的基因型。IL-6、IL-1β、TNF-α基因区段内SNP位点rs1799964、rs1143627及rs1800795等位基因频率在颅内动脉瘤组与正常对照组间具有显著性差异(x2=5.256, P=0.022)(x2=5.521, P=0.019)(x2=6.767, P=0.009)。SNP位点rs1143627及rs1800795等位基因频率与颅内动脉瘤后SAH的病情严重程度相关(χ2=4.257,P=0.039)(χ2=4.083,P=0.043)；此外rs1143627位点还与颅内动脉瘤后SAH的临床预后相关(χ2=5.249,P=0.022)。我们计算了本研究组中TNF-α、IL-1β及IL-6基因中3个SNPs的假阳性报告率,在先验概率为0.25-0.001范围内,3个SNPs的FPRP值均<0.4,且统计检验效能均大于90%,即计算得到的FPRP值均小于预设临界值,因此认为遗传变异与疾病之间的关联是有意义的。4.结论炎症相关因子IL-6、IL-1p、TNF-α基因多态性与颅内动脉瘤易感性相关,为进一步研究IL-6、IL-1p、TNF-α在颅内动脉瘤内作用机制奠定了基础。第二部分炎症相关基因SNP位点与颅内动脉瘤易感性相关分子生物学机制初步研究1.目的探索炎症相关基因SNP位点与颅内动脉瘤易感性相关分子生物学机制,为预防及治疗颅内动脉瘤提供新的思路。2.材料和方法2.1研究对象健康体检者为2013年间于本院体检中心募集的健康体检者24名。年龄>18岁,均为汉族人群,所有受试者均无颅内动脉瘤病史、颅内动脉瘤家族史、脑血管、高血压疾病史、肿瘤史、无脑部外伤史,近期未服用抗炎药物,无心、脑、肝、肾疾病,糖尿病、甲状腺病史,近期无感染及炎症。受试者均经肘静脉取50ml全血,柠檬酸钠抗凝,用于基因表达量的分析和血清学指标的检测。SNP位点为上一章节中验证与颅内动脉瘤相关的多态性位点,即rs1799964、rsl143627及rs1800795。2.2研究方法健康受试者血液样本的采集和PBMCs的分离,并用Trizol提取其中的总RNA,在Nanodrop ND-2000紫外分光光度计上测定其含量和纯度以1%琼脂糖电泳鉴定RNA有无降解,应逆转录试剂盒构建cDNA文库,并用荧光定量PCR检测目的基因1mRNA的表达量。用酶联接免疫吸附技术检测血清中IL-6、IL-1β、TNF-α的含量。血清TNF-α、IL-1β及IL-6mRNA水平及蛋白水平数据的计量资料用均数±标准差表示,经方差齐性检验后(P>0.05)采用单因素方差分析分析,使用P<0.05认为差异有统计学意义。以上分析采用SPSS18.0统计软件完成。3.结果经淋巴细胞分离液分离PBMC之后,我们用Trizol提取了白细胞中的RNA,结果显示RNA质量良好,28S、18S清晰,5S条带微弱,说明总RNA无降解,保证了RNA逆转录的效率及可靠性。通过实时荧光定量PCR技术,检测了携带不同基因型的健康体检中TNF-α基因、IL-1β基因及IL-6基因的表达量。我们发现,TNF-a基因rs1799964位点、IL-1β基因rsl143627位点及IL-6基因rs1800795位点的基因型对各自基因表达量无影响。我们通过实时酶联免疫吸附法,检测了携带不同基因型的健康体检中TNF-α、IL-1β及IL-6蛋白在血清中的表达量。TNF-a基因rs1799964位点、IL-1p基因rs1143627位点及IL-6基因rs1800795位点的基因型对各炎症因子血清蛋白表达量无影响。4.结论TNF-α基因区域内的rs1799964位点与TNF-α mRNA及血清蛋白水平之间无相关性。IL-1β基因区域内的rs1143627位点与IL-1βmRNA及血清蛋白水平之间无相关性。IL-6基因区域内的rs1800795位点与IL-6mRNA及血清蛋白水平之间无相关性。这些SNP位点是通过何种作用发挥其颅内动脉瘤易感性的机制还需进一步的研究。更多还原

【Abstract】 Intracranial aneurysm is local vascular abnormalities, leading to cystic swelling on the intracranial arterial wall, thus creating brain aneurysm formation like protrusions.80%of intracranial aneurysms occur in the first half of the cerebral arterial circle, which has a rupture tendency, once ruptured, it can cause cerebral subarachnoid hemorrhage, thus putting intracranial aneurysms one of the most devastating brain vascular disease. Research shows that85%of SAH due to intracranial aneurysm rupture The mortality rate after aneurysm rupture within24h reach20%, one-week40%, within a month mortality can be up to50%. However, among those survived, over1/3have severe neurological deficits left. Although the incidence of intracranial aneurysms is1%-8%, ranking No.3intracranial lesions, less than cerebral thrombosis, hypertension caused cerebral hemorrhage, its high mortality, high morbidity, all creating a heavy burden to family and societyMultiple factors contributed to intracranial aneurysm, current study suggests that physical labor, mood fluctuation, drinking, etc. can cause sudden rupture of the aneurysm.up to now factors like age, gender, smoking, alcohol consumption, hypertension has been confirmed to be intracranial aneurysm risk factors. Inflammation refers to what the defense system in the living tissue take up against damage factors, vascular reaction is a central part of the inflammatory process. Inflammatory cell mediated abnormal vascular remodeling is considered to be a key aspect of the pathology in a variety of vascular diseases. Changes in blood flow can activate endothelial cells and inflammatory cells releasing inflammatory mediators, increasing vascular wall shear stress can increase the level of IL-1β, facilitating inflammatory cell infiltration into the vessel wall, thus activated endothelial cells, smooth muscle cells jointly mediated inflammatory response. Inflammatory cells, antibody and complement etc. involved in the inflammatory response inducing a variety of proteases which can damage the vessel wall thus causing aneurysm formation. However, until now the specific signaling pathways underlying intracranial aneurysm is not clear. TNF-a released by neutrophils, is a key immunoreactive substance, can strongly activated endothelial cells, it is significantly increased in intracranial aneurysms. In addition, IL-6is also significantly increased in blood and cerebrospinal fluid after the rupture of intracranial aneurysms.Through Gene delivery future generations obtain characteristics similar to parents. DNA as a genetic material through transcription and translation process, and determines the basic characteristics of life, is also closely related to the occurrence of the disease. Genetic factors are important in the pathogenesis of intracranial aneurysms. Epidemiological survey found that some patients with intracranial aneurysms showed onset familial tendency, the risk of the occurrence among the first-degree relatives of patients of the disease was significantly higher. Studies in patients with familial intracranial aneurysm artery among eastern Finnish population found intracranial aneurysms in patients with a positive family history increases the risk rate of12%. Among the Japanese population with family history of intracranial aneurysms, the unruptured aneurysms have a significantly higher prevalence of10.5%-13.5%. Thus, genetic factors play an important role in the occurrence of intracranial aneurysms. SNP refers to genetic marker loci which is a Single nucleotide mutation in the genome. Its large quantity, rich polymorphism contributes to the third generation of genetic markers. Many human phenotypic differences like susceptibility to drugs or disease may be related with the SNP. Although most SNP located in the genome intron elements, but it may play an important role in RNA splicing process. Using whole-genome SNP scans of different races of intracranial aneurysms pedigree confirms a number of chromosomal segments1p34.3-1p36.13、7q11、19ql3.3and Xp22may be associated with the occurrence of aneurysms. In addition, traditional candidate gene study, measureing functionally related genes with intracranial aneurysms, also found a lot of susceptibility genes, such as endothelial glycoprotein gene, matrix metalloproteinase genes, IR collagen gene, otl-antitrypsin gene, elastin gene, angiotensin-converting enzyme gene, apolipoprotein genes. Some of these genes are functionally associated with the formation of the vascular wall, thus creating the molecular evidence. Some studies also suggest that certain inflammatory cytokines like IL-1β, IL-6and TNF-a are positive correlated with intracranial aneurysms susceptibility.Although it has been reported TNF-a, IL-1β, and IL-6are associated with the susceptibility of intracranial aneurysms, they have not been verified in large GWAS samples. In the Italian population, IL-6is not a intracranial aneurysm susceptibility gene. Thus, TNF-a, IL-1β, and IL-6is a susceptibility gene intracranial aneurysms remains uncertain, it may be genetic differences between ethnic groups, or the small number of the sample genome research, lax quality control and not tight statistical analysis strategy leading to an error information. To further confirm that TNF-a, IL-1β, and IL-6gene polymorphism and intracranial aneurysms correlation in Chinese Han population, we collected200cases of intracranial aneurysm patients and200matched peers with strict normal physical examination. The tag SNP loci nearby TNF-a, IL-1(3, and IL-6gene were genotyped, and after strict quality control and rigorous statistical analysis, we try to explore the association of TNF-α, IL-1β, IL6gene polymorphisms with the onset ofntracranial aneurysms among chinese Han population. The implementation of this study will not only made finding the pathogenesis of intracranial aneurysms possible, but also provided important basic molecular biology underlying intracranial aneurysm, facilitate screening among susceptible population and early diagnosis and treatment. Above all, it is of great significance as to raising the detection rate and improving survival and quality of life of patients.Chapter one:Association study of inflammation-related gene polymorphisms and susceptibility of intracranial aneurysms1. ObjectiveTo explore the allele frequencies and genotype frequency distributions of the SNPs in the inflammation-related genes among intracranial aneurysms patients. To investigate the relationship between the inflammation related gene polymorphisms with the severity of intracranial aneurysms after SAH, the clinical relevance of outcomes and the incidence number (solitary/multiple) of intracranial aneurysms.2. Materials and methods2.1Object of study:The168aneurysmal SAH patients with intracerebral hematoma companied or not were choosed as Study group were collected from neurosurgery ward in our NanFang hospital from2010to2012. All Patients choosed were with a onset lower than72hours and older than18years old.they were confirmed by clinical symptoms and then subject to whole brain digital subtraction angiography or CT angiography for the diagnosis of intracranial aneurysms. The control group was184healthy Han people from our hospital’s medical department, all of them had no history of intracranial aneurysms. The three representative SNPs (rs1800795, rs1143627and rs1799964) of tNF-α, IL-1β, and the IL-6gene region were picked out from the NCBI database2.2Research Methods:The clinical data of outpatient intracranial aneurysm patients, hospitalized patients with intracranial aneurysms and healthy Volunteers were all recorded into Excel. Whole blood genomic DNA was extracted using Tigen wholly genomic DNA extraction kit. Then it was tested with agarose gel electrophoresis identified whether it was degraded and measured concentration in the Nanodrop ND-2000spectrophotometer. PCR primers of the corresponding SNP loci were designed according to the nucleotide sequences in NCBI database, then they were used to amplify the sequences which will later used for sequencing in Biotech companies.2.3Statistical analysis:Chi-square test of the SPSS statistical software were used to analysis the allele and genotype frequencies. The allele and genotype frequencies of the SNP loci in TNF-a, IL-1β, and IL-6gene region between cases and controls were compared with the X2test. Mean plus or deviation standard deviation were show for Measurement data, after the homogeneity of variance test (P>0.05), they were compared using ANOVA. P<0.05was considered statistically significant. The progressive model of "Logist regression model" were used to identify the SNP loci of primary effect and assess whether there is a cumulative effect between susceptibility SNP loci and risk factors for intracranial aneurysms. Haploview4.2software were Used for Kazakhstan-Weinberg equilibrium tests and linkage disequilibrium analysis on specific SNP loci segments among the Chinese and Japanese population, LD diagram of the section was drawn.3. ResultsOur study collects168study group intracranial aneurysm patients with male74 cases (44.05%), female94cases (55.95%), mean age (51.67±18.71) years and BMI (23.21±2.16). There were no significant difference between the two groups in general, including history of hypertension, high cholesterol, diabetes, smoking and drinking history (all P>0.05). After treatment the outcomes of the Intracranial aneurysm patients were assessed using the Glasgow Outcome Scale (Glasgow Outcome Scale, GOS), with GOS4～5into a good prognosis and GOS1～3into poor prognosis.122patients with good prognosis (representing72.62%),46patients with poor prognosis (representing27.38%) are found. We first find the the region of the tested genes TNF-a, IL-1β, and IL-6in UCSC library. Then Looking for SNP loci located within the area of10kb range (upstream and downstream5kb) of the three genes in Chinese Han population. At last, we chose rs1799964located upstream of TNF-a gene, rs1143627upstream of the IL-1βgene, rs1800795upstream of the IL-6gene with the most representatives of the region to be examined.The concentration of DNA of whole blood samples among intracranial aneurysms and healthy subjects was between100ng-1μg with good quality. Agarose gel electrophoresis show size about15000kb above with no smearing confirming no degradation. PCR amplification fragmen were shown by agarose gel electrophoresis, all are unity bands with no nonspecific amplification products. The PCR product was genotyped of the SNP loci for each sample. The allele frequency of rs1799964, rs1143627and rs1800795corresponding to IL-6, IL-1β, TNF-a show significant difference between intracranial aneurysm group and normal control group (X2=5.256, P=0.022)(X2=186, P=0.023)(X2=4.368, P=0.037). The allele frequency of rs1143627and rs1800795locus was associated with the severity of illness of intracranial aneurysms after SAH (x2=4.257, P=0.039)(X2=083, P=0.043). In addition rs1143627locus was associated with the clinical outcomes after intracranial aneurysm related SAH.4. Conclusion The polymorphism of inflammatory factor IL-6, IL-1β, TNF-a gene were associated with intracranial aneurysms susceptibility in Chinese Han population. It thus lay foundation for further study of the role IL-6, IL-1β, TNF-αin played in intracranial aneurysm.Chapter two:A preliminary study of the molecular mechanisms of the inflammation related SNP loci which is associated with the of intracranial aneurysms susceptibility in Han population1. ObjectiveExplore molecular mechanisms of the inflammation related SNP loci which is associated with the of intracranial aneurysms susceptibility in Han population and for provide new ideas for the prevention and treatment of intracranial aneurysms.2. Materials and methods2.1Object of study:24healthy volunteers were collected from hospital medical center older than18years old. All are Han population with no history of intracranial aneurysm, intracranial aneurysm family hypertension, cerebrovascular disease, cancer, brain trauma. All did not take anti-inflammatory drugs recently. All with no histroy of cardiovascular, brain, liver, kidney related disease, no diabetes, thyroid disease history, no recent infection and inflammation. Participants were taken50ml whole blood through the cubital vein, sodium citrate anticoagulation, used for analysis gene expression levels and serum markers. SNP loci rs1799964/rs1143627and rs1800795are those associated with intracranial aneurysms which have been verified in the previous section2.2Research Methods:Blood samples from healthy subjects were collected and separated PBMCs, the total RNA were extracted with Trizol. Their content were Measured on a Nanodrop ND-2000spectrophotometer and purity was identified on1%agarose gel electrophoresis to confirm whether the RNA degradated. The expression amount of a reverse transcription to cDNA library was constructed by reverse transcription kit and the target gene mRNA level was mesured by fluorescent quantitative PCR.3. ResultsAfter lymphocyte separation medium by PBMC, we extracted the white blood cells RNA using Trizol, there are28S,18S clear,5S faint bands, indicating no degradation of total RNA, thus ensuring the efficient and reliable reverse transcription. By real-time PCR technology we detect the expression lever of TNF-a, IL-1β, and IL-6gene of the healthy control with different genotypes. We found that the TNF-a gene rs1799964genotype sites, IL-1β gene rs1143627locus rs1800795, and IL-6gene loci have no effect on the expression of each gene. Using real-time enzyme-linked immunosorbent assay we detect the serum protein level of TNF-a, IL-1β and IL-6of the healthy volunteers with different genotypes. TNF-a gene locus rsl799964, IL-1β gene locus rs1143627and IL-6gene locus rs1800795had no effect on the expression of inflammatory cytokines.4. ConclusionNo correlation between the rs1799964locus and TNF-a mRNA and serum TNF-a protein levels. No correlation between the rs1143627locus and IL-1β mRNA and serum IL-1βprotein levels. No correlation between the rs1800795locus and IL-6mRNA and serum IL-6protein levels. What role of these SNP loci to play through in intracranial aneurysms needs further study to confirm it.更多还原