腹泻型肠易激综合征肝郁脾虚证候特点与脑肠神经递质基因多态性的相关性研究

Study on the relationship between the characteristics of irritable bowel syndrome of liver depression and spleen deficiency and the gene polymorphism of brain gut neurotransmitter

[目的]通过全外显子高通量测序技术对腹泻型肠易激综合征(IBS-D)肝郁脾虚证患者进行全外显子测序,初步筛选出与神经递质相关的易感基因,为下一步基因多态性研究奠定基础;分析非共有基因多态性与本病症状的相关性,了解IBS-D分子水平的发病机制。[方法]纳入肝郁脾虚证IBS-D患者,填写临床资料,提取外周血DNA。采用全外显子捕获技术进行高通量测序,测序结果与人类参考基因组序列(UCSC hg19)、avsnp147进行比对过滤。运用ANNOVAR软件对结果进行注释,筛选与神经递质相关的基因,进一步参考1000 Genomes Project、ExAC数据库基因频率注释筛选SNP,整合外显子区域的非同义变异基因,分析本病患者的易感基因,同时探讨非共有基因多态性与本病症状的相关性。[结果]本研究共纳入10例肝郁脾虚证IBS-D患者,测序后每位患者共获得10 G的数据。反复比对过滤测序结果,得到与神经递质相关的基因共5 507个,符合SNP的有5 117个,其中外显子区域非同义变异基因有94个,整合后得到18个共有基因及76个非共有基因。[结论]①rs12437941、rs2402073、rs8187730、rs1390938、rs631357、rs4838544、rs7202563、rs1132356、rs17099370、rs8019076、rs61821060、rs902790、rs564271、rs822522、rs4665855、rs6264、rs2278973、rs10791893这18个基因可能是IBS-D肝郁脾虚证患者的易感基因,但需进一步确认。②IBS-D肝郁脾虚证患者外显子区域与神经递质相关的非同义变异基因多态性有54个与该病症状具有相关性。

[Objective]All exons in patients with diarrhea type irritable bowel syndrome(IBS-D)with liver depression and spleen deficiency were sequenced with high throughput sequencing,while susceptible genes related to neurotransmitters were preliminary screened,laying foundations for next steps of gene polymorphism research.In order to understand pathogenesis of IBS-D at molecular level,To analyze the correlation between the gene polymorphisms and the symptoms of the disease.[Methods]IBS-D patients with liver depression and spleen deficiency were recruited to extracted DNA from peripheral blood with filled in clinical data High throughput sequencing were conducted with total exon capture,while the sequencing results were compared with human reference genome sequence(UCSC hg19)and avsnp147.To annotate results via annovar software,genes related to neurotransmitters were screened,further referred to 1000 genes project,exac database gene frequency annotation to screen SNP.Non synonymous variation gene in exon region were integrated,susceptible genes in patients with this disease were analyzed and at the same time to explore the correlation between the non shared gene polymorphism and the symptoms of the disease.[Results]In this study,10 IBS-D patients with liver stagnation and spleen deficiency were included.a total of 5507 genes related to neurotransmitters were obtained,5117 were in accordance with SNP,94 were exon region non synonymous variation genes.After integration,18 common genes and 76 non common genes were obtained.[Conclusion]①rs12437941,rs2402073,rs8187730,rs1390938,rs631357,rs4838544,rs7202563,rs1132356,rs17099370,rs8019076,rs61821060,rs902790,rs564271,rs822522,rs4665855,rs6264,rs2278973,rs10791893--these 18 polymorphisms may be susceptible genes in IBS-D patients with syndrome of liver depression and spleen deficiency,but it needs further confirmation.②In IBS-D patients with syndrome of liver stagnation and spleen deficiency syndrome,there are 54 non synonymous gene polymorphisms related to neurotransmitters in the exon region,which are related to the symptoms of IBS-D.