NOD2/CARD15基因多态性与克罗恩病患者相关性研究

目的NOD2/CARD15基因是人类的第一个克罗恩病(CD)易感基因,其间的3个单核苷酸多态性(SNPs)与白种人CD有显著性相关,但与日本人无关。本研究旨在证实这3个SNPs是否与浙江地区人群的CD易感性有关。方法血样来自浙江地区32例CD患者,110例溃疡性结肠炎患者及292例健康对照者。通过PCRSSP方法直接检测野生型及NOD2/CARD15基因的3个多态性(Arg702Trp,Gly908Arg,Leu1007fsinsC)。结果没有发现1例CD患者纯合子或杂合子的SNPs突变,同样在溃疡性结肠炎患者和健康人中也未能检测到。结论本研究表明一些存在于特定人群的CD易感基因可能在其他人群中不存在,白种人CD患者相关的易感基因NOD2/CARD15常见的3个SNPs与浙江地区CD人群无关。     

NOD2/CARD15与克罗恩病相关性研究进展

克罗恩病(CD)是一种常见的炎症性肠病,是环境因素作用于易感个体产生的肠道慢性非特异性炎症。其发病机制尚未完全明了。NOD2/CARD15是目前确认的第一个与CD相关的基因,通过介导凋亡,调节免疫等多种途径参与CD的发病。本文就NOD2/ CARD15与CD的研究进展作一综述。     

克罗恩病CARD15/NOD2基因突变的研究

目的 发现与中国人克罗恩病（CD）发病相关的单核苷酸多态性（SNP）及其与临床特点的关系.方法 临床资料完整的CD、溃疡性结肠炎（UC）患者及健康体检者各30例.提取人血白细胞基因组DNA,PCR扩增NOD2基因第4、8、11外显子,纯化后直接测序.结果 5例CD患者有SNP改变,其中2例为P268S,1例为R459R,2例为P268S和R459R,而在UC患者和健康人中各检测到1例R459R.所有研究对象未发现R702W、G908R及3020insC改变.CD有4例P268S,与UC和健康体检者比较差异有统计学意义（χ^2=8.037,P＜0.05）.4例P268S CD患者病变均在回肠（χ^2=9.231,P=0.01）,发病年龄小于20岁（χ^2=10.769,P＜0.01）,并发肠腔狭窄而需手术（χ^2=7.972,P＜0.01）,2例有P268S和R459R的患者病情较重,多次复发.结论 P268S可能是CARD15/NOD2基因中与中国人CD相关的SNP,与患者发病年龄、病变部位及肠腔狭窄明显相关,与患者性别及病变严重程度无关.     

CARD15/NOD2基因多态性与克罗恩病发病的荟萃分析

克罗恩病（CD）是一种具有遗传背景的多因素疾病。位于第16号染色体上的CARD15／NOD2基因被确定为克罗恩病的易感基因，该基因是参与细胞凋亡调控的Apaf21／PCed4超家族成员之一，主要在单核巨噬细胞内表达。有研究证明，CARD15／NOD2基因上三个多态位点（R702W、G908R、3020sinC）与克罗恩病发病相关。为综合评价CARD15／NOD2基因序列单核苷酸多态性同克罗恩病发病的相关性，我们用荟萃分析方法对二者的关系进行综合定量再分析，探讨CARD15／NOD2基因多态性与克罗恩病发病的关系。     

NOD2/CARD15基因突变与中国人克罗恩病相关性的研究

背景：近年多项研究证明NOD2／CARD15基因序列的单核苷酸多态性（SNP）与西方白种人克罗恩病（CD）明显相关．其中3个SNP（R702W、G908R和3020ins C）与CD的相关性尤为显著。目的：探讨NOD2／CARD15基因SNP与中国人CD发病的相关性及其与CD临床特点的关系。方法：选取临床资料完整的CD患者48例、溃疡性结肠炎（UC）患者和健康对照者各50例，提取人血白细胞基因组DNA，经聚合酶链反应（PCR）扩增NOD2基因全部12对外显子，纯化后直接测序，根据结果分析其突变与CD病变特点的关系。结果：CD组、UC组和健康对照组均未检出3个西方人常见的NOD2／CARD15基因多态性位点。CD组的P268S突变率显著高于UC组和健康对照组（P〈0．05）。5例P268S突变的CD患者病变均位于回肠（P〈0．01），4例发病年龄≤20岁（P〈0．01），且均并发肠腔狭窄（P〈0．01）。结论：中国人CD患者中存在NOD2／CARD15基因P268S突变，且与患者的发病年龄、病变部位和并发症相关，有必要对其功能作进一步探讨。     

ATG16L1基因多态性与广西壮族人群炎症性肠病的关系

目的研究ATG16L1基因单核苷酸多态性（SNP）位点rs2241880与中国广西壮族炎症性肠病（IBD）易感性的关系。方法分别收集146例中国广西IBD患者[其中溃疡性结肠炎（UC）86例,克罗恩病（CD）60例]与164例正常对照者的肠黏膜组织,用酚一氯仿法提取DNA,采用聚合酶链反应（PCR）扩增ATG16L1基因,PCR产物片段直接进行DNA测序分析,结果在Genbank基因库上进行序列比较。结果UC、CD患者与正常对照组之间ATG16L1基因SNP位点rs2241880的等位基因多态性无统计学差异（P〉0．05）。结论ATG16L1基因SNP位点rs2241880与广西壮族IBD患者的易感性无关。     

克罗恩病的第一个易感基因,NOD2(CARD15)

炎症性肠病 (ＩＢＤ)具有遗传易感性 ,主要表现在家族聚集现象和双胎同胞的共患率[1] 。近年来 ,随着人类基因组和多基因疾病研究以及统计学方法的进展 ,相继发现ＩＢＤ的基因易感位点 ,证实ＩＢＤ的遗传易感性涉及多个基因位点[2 ] 。 1996年Ｈｕｇｏｔ等[3 ] 采用非参数连锁分析 (ｎｏｎｐａｒａｍｅｔｒｉｃｌｉｎｋａｇｅａｎａｌｙｓｉｓ,ＮＰＬ)法 ,对ＩＢＤ患病同胞 (ａｆｆｅｃｔｅｄｓｉｂｌｉｎｇｐａｉｒｓ,ＡＳＰ)进行基因组位点微卫星标记研究 ,发现克罗恩病 (ＣＤ)的易感位点位于第 16条染色体的着丝点附近 ,称为ＩＢＤ1位点 …     

IL-23R基因多态性与炎症性肠病的相关性

目的:研究我国炎症性肠病患者IL-23R基因单核苷酸多态性特征,探讨其与炎症性肠病(IBD)的相关性.方法:采用PCR扩增和测序的方法对198例IBD患者和100名健康体检者(对照组)IL-23R基因3个非同义单核苷酸多态性(rs11209026,p.Arg381Gln;rs41313262,p.Val362Ile;rs11465797,p.Thr175Asn)进行分析,分别计算其等位基因的表现频率,并结合临床资料评估其多态性与IBD的相关性.结果:(1)IBD中克罗恩病(CD)患者IL-23RArg-381Gln等位基因A的表现频率为2.70%,低于对照组(6.00%),但无统计学差异,溃疡性结肠炎(UC)患者的表现频率为5.65%,与对照组无统计学差异;(2)IBD中CD患者IL-23RVal-362Ile等位基因A的表现频率为2.70%,UC组为2.42%,对照组为2.00%,3组间无明显统计学差异;(3)IBD及对照组中均无Thr175Asn等位基因A表现,相应位点均表现为C.结论:我国IL-23R单核苷酸多态性与IBD无明显相关性,遗传异质性可能决定了IBD的易感性.     

中国人群炎症性肠病易感基因多态性的研究进展

炎症性肠病（IBD）是一种多基因复杂疾病,其家族聚集性及同卵双生子高共患率的现象均提示遗传因素在IBD的发病中发挥重要作用.而不同的单核苷酸基因多态性（SNP）对IBD易感性的影响存在明显的种族、地区及人群差异性.近来国外研究显示,欧洲人群IBD的易感基因/位点多达99个,其中与克罗恩病（CD）及炎症性肠病（UC）均相关的有28个.过去一直认为IBD好发于西方国家,但近20年来,随着消化内镜技术的发展,亚洲及中东地区IBD的检出率明显增高,国内对基因多态性与IBD易感性关系的研究也逐渐增多,但与西方国家比较存在明显的东西差异.现对中国人群炎症性肠病易感基因多态性的研究进展综述如下.     

NAT2基因多态性与温州汉族人群炎症性肠病遗传易感性的相关性

目的:探讨温州汉族人群中炎症性肠病(IBD)遗传易感性与N-乙酰基转移酶2(NAT2)基因型多态性的相关性.方法:采用聚合酶链反应-限制性片断长度多态性方法,在119例IBD患者及120例健康对照者中,检测NAT2野生型等位基因(NAT2 4)和3种突变型等位基因(NAT2 5B,6A和7B)的频率.结果:在IBD组中,NAT2 4,NAT2 5B,NAT26A和NAT2 7B等位基因频率分别是55.9%,6.7%,23.5%和13.9%,与正常对照组比较无显著差异.CD组和UC组中各等位基因频率与正常对照组比较无显著差异;将NAT2基因型分为快型、中间型和慢型,分别为35.3%,41.2%和23.5%,与正常对照组比较亦无显著差异;对IBD各组进一步分层,也无显著性差异.结论:NAT2基因型多态性和炎症性肠病遗传易感性无显著性相关.     

Association between polymorphisms in the Toll-like receptor 4, CD14, and CARD15／NOD2 and inflammatory bowel disease in the Greek population

AIM: Crohn's disease (CD) and ulcerative colitis (DC) are multifactorial diseases with a significant genetic background. Apart from CARD15/NOD2 gene, evidence is accumulating that molecules related to the innate immune response such as CD14 or Toll-like receptor 4 (TLR4), are involved in their pathogenesis. In further exploring the genetic background of these diseases, we investigated the variations in the CARD15/NOD2 gene (Arg702Trp, Gly908Arg and Leu1007fsinsC), and polymorphisms in the TLR4 gene (Asp299Gly and Thr399Ile) as well as in the promoter of the CD14 gene (T/C at position -159) in Greek patients with CD and UC. METHODS: DNA was obtained from 120 patients with CD, 85 with UC and 100 healthy individuals. Genotyping was performed by allele specific PCR or by PCR-RFLP analysis. RESULTS: The 299Gly allele frequency of the TLR4 gene and the T allele and TT genotype frequencies of the CD14 promoter were significantly higher in CD patients only compared to healthy individuals (P= 0.026<0.05; P= 0.0048<0.01 and P= 0.047<0.05 respectively). Concerning the NOD2/CARD15 mutations the overall presence in CD patients was significantly higher than that: in UC patients or in controls. Additionally, 51.67% of the CD patients were carriers of a TLR4 and/or CD14 polymorphic allele and at least one variant of the NOD2/CARD15, compared to 27% of the UC patients. It should be pointed out that both frequencies significantly increased as compared with the 10% frequency of multiple carriers found in healthy controls. A possible interaction of the NOD2/CARD15 with TLR4 and especially CD14, increased the risk of developing inflammatory bowel disease (IBD). CONCLUSION: Our results indicate that co-existence of a mutation in either the TLR4 or CD14 gene, and in NOD2/CARD15 is associated with an increased susceptibility to developing CD compared to UC, and to developing either CD or UC compared to healthy individuals.     

Risk factors and gene polymorphisms of inflammatory bowel disease in population of Zhejiang, China

AIM:To identify the risk factors and three single nucleotide polymorphisms(SNPs) of NOD2/CARD15 gene in inflammatory bowel disease(IBD) of the population in Zhejiang,China.METHODS:A case-control study was conducted using recall questionnaire to collect data on demographic,socioeconomic,lifestyle characteristics and dietary behaviors from 136 determined IBD patients and 136 paired healthy controls.COX regression method was used to screen the statistically significant risk factors for IBD.The polymorphisms of...     

Clinical significance of NOD2/CARD15 and Toll-like receptor 4 gene single nucleotide polymorphisms in inflammatory bowel disease

AIM： To evaluate the role of genetic factors in the pathogenesis of Crohn＇s disease （CD） and ulcerative colitis （UC）, we investigated the single nucleotide polymorphisms （SNPs） of NOD2/CARD15 （R702W, Gg08R and L1007finsC）, and Toll-like receptor 4 （TLR4） genes （D299G and T399I） in a selected inflammatory bowel disease （IBD） population coming from Southern Italy. METHODS： Allele and genotype frequencies of NOD2/ CARD15 （R702W, Gg08R and L1007finsC） and TLR4 （D299G and T399I） SNPs were examined in 133 CD patients, in 45 UC patients, and in 103 healthy controls. A genotype-phenotype correlation was performed. RESULTS： NOD2/CARD15 R702W mutation was significantly more frequent in CD （9.8%） than in controls （2.4%, P = 0.001） and in UC （2.3%, P = 0.03）. No significant difference was found between UC patients and control group （P 〉 0.05）. In CD and UC patients, no significant association with G908R variant was found. L1007finsC SNP showed an association with CD （9.8%） compared with controls （2.9%, P = 0.002） and UC patients （2.3%, P = 0.01）. Moreover, in CD patients, G908R and L1007finsC mutations were significantly associated with different phenotypes compared to CD wild-type patients. No association of IBD with the TLR4 SNPs was found in either cohort （allele frequencies： D299G-controls 3.9%, CD 3.7%, UC 3.4%, P 〉 0.05; T399I-controls 2.9%, CD 3.0%, UC 3.4%, P 〉 0.05）. CONCLUSION： These findings confirm that, in our IBD patients selected from Southern Italy, the NOD2/ CARD15, but not TLR4 SNPs, are associated with increased risk of CD.     

1007fs, G908R, R702W Mutations and P268S, IVS8+158 Polymorphisms of the CARD15 Gene in Turkish Inflammatory Bowel Disease Patients and Their Relationship with Disease-Related Surgery

Introduction CARD15 gene mutations may present different frequencies in populations and sometimes surgical interventions may become a necessary therapy for inflammatory bowel disease patients. Mutations of 1007fs, G908R, R702W and polymorphisms of P268S, IVS8⁺¹⁵⁸ of the CARD15 gene and their relation with disease-related surgery were investigated in Turkish inflammatory bowel disease patients in this study. Material and Method 1007fs, G908R, R702W mutations and P268S, IVS8⁺¹⁵⁸ polymorphisms of CARD15 gene were analyzed in 130 inflammatory bowel disease patients (67 Crohn’s disease, 63 ulcerative colitis) and 87 healthy controls. After obtaining DNA samples, genotyping was performed by polymerase chain reaction - restriction fragment length polymorphism (PCR-RFLP) analysis. Results were evaluated by statistical analysis and accepted as significant if P < 0.05. Results R702W gene mutation was significantly lower in the inflammatory bowel disease group (1.5%) than the controls (4.8%) (P < 0.05). The overall allele frequency of mutations in the inflammatory bowel disease group (2.7%) was lower than in controls (6.6%) (P < 0.05). Disease-related surgery history was present in 20 Crohn’s and 25 ulcerative colitis patients; familial history was present in four Crohn’s and five ulcerative colitis patients. Statistically, no relationship was detected between disease-related surgeries and the investigated genetic tests. Conclusion In Turkish patients, no important relationship was detected between the investigated allele frequencies of the CARD15 gene and inflammatory bowel disease nor between disease-related surgeries and inflammatory bowel disease. Dedicated to the memory of the Turkish scientist Turgut Tukel MD. Thanks for his contributions and supports.     

NOD2/CARD15 , ATG16L1 and IL23R gene polymorphisms and childhood-onset of Crohn’s disease

AIM: To assess whether the polymorphisms of NOD2/ CARD15 , autophagy-related 16-like 1 (ATG16L1 ), and interleukin-23 receptor (IL23R ) genes play a more critical role in the susceptibility of childhood-onset than in adult-onset Crohn’s disease (CD). METHODS: Polymorphisms R702W, G908R, and 3020insC of NOD2/CARD15 ; rs2241880 A/G of ATG16L1 , and rs11209026 (R381Q) of IL23R gene were assessed in 110 childhood-onset CD, 364 adult-onset CD, and 539 healthy individuals. Analysis of polymorphisms R702W, G908R, ...     

Association between NOD2/CARD15 gene polymorphisms and Crohn’s disease in Chinese Zhuang patients

AIM:To assess the relationship between the P268S,JW1 and N852S polymorphisms and Crohn’s disease(CD)susceptibility in Zhuang patients in Guangxi,China.METHODS:Intestinal tissues from 102 Zhuang[48CD and 54 ulcerative colitis(UC)]and 100 Han(50 CD and 50 UC)unrelated patients with inflammatory bowel disease and 72 Zhuang and 78 Han unrelated healthy individuals were collected in the Guangxi Zhuang Autonomous Region from January 2009 to March 2013.Genomic DNA was extracted using the phenol chloroform method.The P268S,JW1 and N852S polymorphisms were amplified using polymerase chain reaction(PCR),detected by restriction fragment length polymorphism(RFLP),and verified by gene sequencing.RESULTS:Heterozygous mutation of P268S in the NOD2/CARD15 gene was detected in 10 CD cases(six Zhuang and four Han),two Han UC cases,and one Zhuang healthy control,and P268S was strongly associated with the Chinese Zhuang and Han CD populations(P=0.016 and 0.022,respectively).No homozygous mutant P268S was detected in any of the groups.No significant difference was found in P268S genotype and allele frequencies between UC and control groups(P>0.05).Patients with CD who carried P268S were likely to be≤40 years of age(P=0.040),but were not significantly different with regard to race,lesion site,complications,and other clinical features(P>0.05).Neither JW1 nor N852S polymorphisms of the NOD2/CARD15gene were found in any of the subjects(P>0.05).CONCLUSION:P268S polymorphism may be associated with CD susceptibility in the Zhuang population in the Guangxi Zhuang Autonomous Region,China.In contrast,JW1 and N852S polymorphisms may not be related to CD susceptibility in these patients.     

CARD15/NOD2 polymorphisms do not explain concordance of Crohn''s disease in Swedish monozygotic twins

BACKGROUND: CARD15/NOD2 polymorphisms are associated with Crohn's disease. There is a high concordance for disease and disease phenotype in monozygotic twin pairs with Crohn's disease. AIM: We studied CARD15/NOD2 polymorphisms in a Swedish, population-based cohort of monozygotic twins with Crohn's disease to assess whether these variants explain disease concordance. SUBJECTS AND METHODS: Twenty-nine monozygotic twin pairs (concordant n=9, discordant n=20) with Crohn's disease and 192 healthy controls were investigated for the CARD15/NOD2 variants Arg702Trp, Gly908Arg and Leu1007fsinsC. RESULTS: CARD15/NOD2 mutations were found in 5/38 (13%) twins with Crohn's disease, corresponding to a total allele frequency of 6.6%. Only 2/9 concordant twin pairs carried any of the variants and the remaining seven were wild type genotype. The total allele frequency was 4.4 times higher (95% confidence interval 1.0-21.5, p=0.06) in concordant twins than in discordant ones, 11.1% versus 2.5%. In healthy controls the total allele frequency was 2.6%. CONCLUSIONS: CARD15/NOD2 polymorphisms contribute but do not alone explain concordance of Crohn's disease in monozygotic twins and, at least in a Swedish population, other polymorphisms are required. The low occurrence of CARD15/NOD2 mutations in the study and other Northern European populations suggests that these variants are of less importance in Northern Europe.     

OCTN and CARD15 gene polymorphism in Chinese patients with inflammatory bowel disease

AIM： To investigate the single nucleotide polymorphism （SNPs） distribution of NOD2/CARD15 （R702W, G908R）, OCTN1 1672CFT and OCTN2-207G/C in Chinese patients with inflammatory bowel disease （IBD）. METHODS： A total of 61 patients with Crohn＇s disease （CD）, 151 patients with ulcerative colitis （UC）, and 200 unrelated healthy controls were genotyped. Genotyping was performed by sequence specific primer polymerase chain reaction （PCR-SSP） or by restriction fragment length polymorphism （PCR-RFLP） analysis. RESULTS： Among the subjects in our study groups, including patients with CD, UC and healthy controls, none had OCTN and CARD15 variants and very rare IBD family history was found in our patients with the percentage of 0 （0/61 with CD） and 1.3% （2/151 with UC）. CONCLUSION： Our results indicate that although OCTN or CARD15 variation is associated with susceptibility to IBD in Western populations, these might be rare and may not be associated with susceptibility to IBD in Chinese patients.