【www.zhangdahai.com--开题报告】
[摘要] 目的 探讨5-羟色胺转运体基因多态性与强迫症发病的相关性。方法 采用聚合酶链反应扩增技术测定强迫症患者和健康人的SLC6A4基因型。结果 强迫症组SLC6A4第2内含子的基因型及等位基因分布与对照组间的差异有统计学意义(P0.05)。结论 SLC6A4基因可能与强迫症存在遗传关联,第2内含子的10和12/10基因型、启动子的L/L基因型可能是强迫症的风险因子。
[关键词] 强迫性障碍;基因;多态性;限制性片段长度;5-羟色胺转运体
[中图分类号] R749.5 [文献标识码] A[文章编号] 1673-9701(2010)02-09-03
Association between 5- Serotonin Transporter Gene Polymorphism and Obsessive-Compulsive Disorder
FENG Dongmei1GAN Luchun1LIANG Songyou1LIU Jinying1DAI Juan1SHI Qiao1HAN Zili2TAO Jiong2
1.Psychology Department,Zhongshan Municipal Central Hospital,Zhongshan 528400,China;2. Psychology Department,the Third Affiliated Hospital of Zhongshan University,Guangzhou 510000,China
[Abstract] Objective To explore the association between 5- serotonin transporter gene polymorphism(SLG6A4) and obsessive-compulsive disorder(OCD). Methods Altogether 50 unrelated OCD patients and 60 sex-matched healthy controls were included in this study and all the subjects were genotyped directly by using polymerase chain reaction amplification fragment length polymorphism techniques. Results There were significant differences between patients and controls in the SLC6A4 intron 2 genotype and allele distribution(χ2=10.42, P=0.005; χ2=6.061, P=0.048). Allele 10 and genotype 12/10 of intron 2 and genotype L/L of promoter of SLC6A4 were positively associated with OCD. There was no significant difference in the genotype of SLC6A4 in the sex between the two groups.Conclusion The polymorphism of SLC6A4 gene may be associated with OCD,and its allele 10 and the genotype 12/10 intron 2 or genotype L/L of promoter may be risk factors for OCD.
[Key words] Obsessive-compulsive disorder;Gene;Polymorphism;Reaction fragment length;Serotonin transporter
强迫症(obsessive-compulsive disorder,OCD)是一种慢性精神障碍性疾病,其主要症状为反复出现的强迫观念和强迫动作,患者明知道没意义却无法摆脱;强迫症的终生患病率为2%～3%,有超过10%的强迫症患者对系统规范的药物治疗和行为治疗无效或效果不佳。强迫症由于其难治性和高致残性而引起人们的重视。至今OCD的病因尚不明确,随着分子遗传学技术的进步,基因在OCD的发病中的作用日益受到重视[1],许多研究显示基因在强迫症的发病中可能具有重要作用并提出了大量“候选基因”,近年来5-羟色胺再摄取抑制剂(SRIs)治疗OCD取得了部分的成功,而5-羟色胺转运体(5-HTT)正是SRIs药物发挥作用的靶结构,编码该转运体的SLC6A4基因便成为OCD遗传病因学研究中的热门基因。许多研究者认为,该基因的第2内含子区的可变数目串联重复(variable number tandem repeat,VNTR)多态以及启动子区的44 bp的插入/缺失多态可能与OCD的发生有关,为探讨该多态性与OCD的关系,我们对OCD患者和健康人的SLC6A4基因多态性进行了检测。
1材料与方法
1.1对象及分组
1.1.1强迫症(OCD)组 我院2005年9月~2008年9月的门诊和住院OCD患者50例,均符合中国精神障碍分类与诊断标准第3版及国际疾病分类第10版关于强迫症的诊断标准,均无器质性疾病;入组前未服抗精神病药,如服药则予2周清洗期;汉族;Yale-Brown强迫量表(Y-BOCS)总分均≥16分。50例中男30例,女20例,平均发病年龄为(31±9)岁。
1.1.2对照组为同期本院职工、进修医生和大学生志愿者60名,年龄18~60岁,无重大躯体疾病,无任何符合诊断标准的精神疾病,既往无精神疾病史,无精神疾病家族史。60名中男32名,女28名;平均年龄(33±11)岁。
所有研究对象均无血缘关系,均对本次研究知情及同意。强迫症组和对照组一般资料相匹配。
1.2方法
1.2.1DNA的提取抽取静脉血2mL,EDTA抗凝;分离白细胞后采用常规方法提取DNA。
1.2.2SLC6A4基因的聚合酶链反应(PCR)扩增引物序列见参考文献[2,3]。反应体积25μL,成分:500 ng基因组DNA,20 pmol引物,200 mmol/LdNTPs,1×PCR缓冲液,1U Taq酶。第2内含子区(5-HTTVNTR)PCR反应程序为:94℃ 4min。30×(94℃,30s;58℃,40s;72℃,60s),72℃ 10min;启动子区(5-HTTLPR)PCR反应程序为95℃ 3min,32×(95℃,20s;60℃,20s;72℃,40s),72℃ 10min。
1.2.3电泳分型(1)5-HTTVNTR电泳分型。12% PAGE 180 V电泳90 min,溴化乙啶(EB)染色20 min,拍照分型。267 bp为等位基因10,301 bp为等位基因12。(2)5-HTTLPR电泳分型。3%琼脂糖凝胶电泳100 V电泳100 min,EB染色2O min,拍照分型。367 bp为等位基因S,411 bp为等位基因L。
1.3统计学方法
全部数据输入计算机,使用SPSS13.0统计软件进行数据分析。方法包括频数分析、Hardy-Weinberg平衡法则(H-W定律)的吻合度检验、χ2检验等。
2结果
2.1OCD组和对照组的H-W定律吻合度检验
OCD组观察值与期望值吻合良好(χ2=2.71,v=1,P>0.05);对照组观察值和期望值吻合良好(χ2=2.86,v=1,P>0.05);均符合H-W定律。
2.2OCD组与对照组间SLC6A4基因型及等位基因频数分布的比较
见表1,由表1可见,OCD组与对照组的SLC6A4第2内含子基因型(χ2=10.42,P0.05)和第2内含子(χ2=0.50,P>0.05)的基因型分布、对照组内不同性别间SLC6A4启动子(χ2=2.64,P>0.05)和第2内含子(χ2=1.87,P>0.05)基因型分布的差异均无显著性。
2.4SLC6A4基因型与OCD的关联分析
关联分析结果显示,OCD患者与5-HITVNTR等位基因l0[比数比(OR)=2.24,Z=2.69,P
[参考文献]
[1] Meira-Lima I,Shavitt RG,Miguita K,et al. Association analysis of the catechol-o-methyltransferase(COMT),serotonin transporter(5-HTT)and serotonin 2A receptor(5HT2A)gene polymorphisms with obsessive- compulsive disorder[J].Genes Brain Behav,2004,3(2):75-79.
[2] MacKenzie A,Quinn J. A serotonin transporter gene intron 2 polymorphic region,correlated with affective disorders,has allele-dependent differential enhancer-like properties in the mouse embryo[J]. Proc Natl Acad Sci USA,1999,96(26):15251-15255.
[3] Ramoz N,Reichert JG,Corwin TE,et al. Lack of evidence for association of the serotonin transporter gene SLC6A4 with autism[J]. Biol Psychiatry,2006,60(2):186-191.
[4] McDougle CJ,Epperson CN,Price LH,et al. Evidence for linkage disequilibrium between serotonin transporter protein gene(SLC6A4)and obsessive compulsive disorder[J]. Mol Psychiatry,1998,3(3):270-273.
[5] Camarena B,Rinetti G,Cruz C,et al. Association study of the serotonin transporter gene polymorphism in obsessive-compulsive disorder[J]. Int J Neuropsychopharmacol,2001,4(3):269-272.
[6] Denys D,Van Nieuwerburgh F,Deforce D,et al. Association between serotonergic candidate genes and specific phenotypes of obsessive compulsive disorder[J]. J Affect Disord,2006,91(1):39-44.
[7] Baca-Garcia E,Vaquero-Lorenzo C,Diaz-Hernandez M,et al. Association between obsessive-compulsive disorder and a variable number of tandem repeats polymorphism in intron 2 of the serotonin transporter gene[J]. Prog Neuropsychopharmacol Biol Psychiatry,2007,31(2):416-420.
[8] Hayden EP,Dougherty LR,Maloney B,et al. Temperamental fearfulness in childhood and the serotonin transporter promoter region polymorphism:a multimethod association study[J]. Psychiatr Genet,2007,17(3):135-142.
[9] Heils A,Teufel A,Petri S,et al.Functional promoter and polyadenylation site mapping of the human serotonin(5-HT)transporter gene[J]. J Neural Transm Gen Sect,1995,102(3):247-254.
(收稿日期:2009-06-12)
