五羟色胺转运体基因及神经肽S受体基因多态性与原发性失眠的致病机制研究

The suffering of primary insomnia (PI) may has a genetic basis. The short (s-) allele of the 5-HTT gene-linked polymorphic region (5-HTTLPR) has been reported associated with insomnia. The neuropeptide S receptor (NPSR) and its ligand neuropeptide S (NPS) was reported to be involved in arousal regulation. Previous studies have suggested that the NPSR rs324981 polymorphism is associated with decreased sleep duration and extended sleep lantency. It suggests that the gene polymorphism of 5-HTT and NPSR may result in PI. This project aims to explore the association between the NPSR, 5-HTT gene polymorphism and PI in China via the denaturing high performance liquid chromatography (DHPLC) and direct DNA sequencing in combination with pittsburgh sleep quality index (PSQI) ,insomnia severity index (ISI) and polysomnography (PSG), and the correlations between the founded polymorphic loci and objective sleep phenotype (sleep duratoin, sleep lantency, N3%, REM%, microarousal index, sleep efficiency, etc.) or subjective sleep phenotype (sleep lantency, sleep time, night time waking times, etc.). The project is expected to enrich the pathogeny of PI theory and adopt NPSR and 5-HTT gene polymorphism as possible reference indexes for clinical PI diagnosis and treatment.

原发性失眠（PI）有一定的遗传特征。研究表明编码五羟色胺转运体（5-HTT）基因的连锁多态区（5-HTTLPR）S 等位基因可能与失眠相关。新发现的神经肽S（NPS）及其受体（NPSR）系统参与睡眠觉醒调节，NPSR基因多态性（NPSR rs324981）与人类睡眠潜伏期延长和睡眠时间缩短密切相关，提示5-HTT基因多态性及NPSR基因多态性与PI有一定的关联性。本项目拟通过DHPLC及DNA直接测序技术，联合PSQI、ISI量表评价、PSG监测，揭示PI中国人群中5-HTT基因及NPSR基因多态性与PI的关联性，并揭示各多态位点与客观睡眠表型（睡眠时间、潜伏期、N3%、REM%、微觉醒指数、睡眠效率等）及主观睡眠表型（睡眠时间、睡眠潜伏期、夜间苏醒次数等）的关联性。旨在探索NPSR与5-HTT基因多态性检测能成为PI诊断重要可能指标，为PI诊断和治疗提供理论基础。

研究背景：原发性失眠（PI）有一定的遗传特征。研究表明编码五羟色胺转运体（5-HTT）基因的连锁多态区（5-HTTLPR）S 等位基因可能与失眠相关。新发现的神经肽S（NPS）及其受体（NPSR）系统参与睡眠觉醒调节，NPSR基因多态性（NPSR rs324981）与人类睡眠潜伏期延长和睡眠时间缩短密切相关，提示5-HTT基因多态性及NPSR基因多态性与PI有一定的关联性。本项目拟通过DHPLC及DNA直接测序技术，联合PSQI、ISI量表评价、PSG监测，揭示PI中国人群中5-HTT基因及NPSR基因多态性与PI的关联性，并揭示各多态位点与客观睡眠表型（睡眠时间、潜伏期、N3%、REM%、微觉醒指数、睡眠效率等）及主观睡眠表型（睡眠时间、睡眠潜伏期、夜间苏醒次数等）的关联性。旨在探索NPSR与5-HTT基因多态性检测能成为PI诊断重要可能指标，为PI诊断和治疗提供理论基础。.主要研究内容:利用生物信息学方法和国际HapMap计划以及NCBI公布的人群中的单核苷酸多态性（single nucleotide polymorphism, SNP）数据，构建出单体型域，并结合功能提示，共挑选4个5-HTT及13个NPSR等位基因频率>5%且符合哈-温平衡的单体型标签SNP位点。共收集到157例原发性失眠症患者，对其4个5-HTT及13个NPSR标签SNP位点进行测序，并对133例年龄、性别相匹配的正常对照组患者的相同位点进行测序。.重要结果及关键数据:所有SNP位点基因型在对照组中符合Hardy-Weinberg平衡检测(p>0.05)。失眠组与对照组的年龄、性别无统计学意义(均p>0.05)。NPSR的SNP位点rs323920的T/C型与健康对照组比较有统计学意义(p=0.043)， NPSR的SNP位点rs324957的A/G型与健康对照组比较有统计学意义(p=0.015)。.科学意义:神经肽S基因多态性rs323920和rs324987可能是原发性失眠的易感基因位点，NPSR的SNP单体型rs323920的基因型T >C变异和rs324957基因型的A>G变异可能是原发性失眠的易感基因。