先天性小耳畸形差异性代谢物质及代谢通路的非靶向筛查

Non-targeted metabonomics study on congenital microtia based on ear cartilage

目的筛查先天性小耳畸形患者残耳软骨组织中相关差异性代谢物和关键的代谢通路。方法根据纳入标准,选取2017年6月至2018年1月中国医学科学院北京协和医学院整形外科医院收治的单纯性小耳畸形患者18例为病例组,行耳再造术时采集患者的残耳软骨组织。另选取18例非小耳畸形、但治疗过程需采取耳软骨的患者为对照组,采取正常耳廓软骨组织。将样本预处理后,采用气相色谱飞行时间质谱联用技术(GC-TOFMS)的非靶向代谢组学的研究手段,XploreMET软件行代谢组学数据分析,利用正交偏最小二乘法判别分析(OPLS-DA)及其变量重要性投影(VIP)法和U检验的统计方法,分析小耳畸形残耳软骨组织中的差异性代谢物,并将其比照代谢网络数据库,利用代谢通路富集分析(MPEA)筛选得出与差异性代谢物最为相关的代谢通路。结果小耳畸形患者的残耳软骨组织和正常耳廓软骨组织中的代谢产物具有差异,其中14个代谢物差异具有统计学意义(P<0.05);3条代谢通路可能发生显著改变(P<0.05),为精氨酸合成途径、牛磺酸和亚牛磺酸代谢途径、维生素B5和辅酶A合成途径。结论精氨酸、牛磺酸、L-半胱氨酸与小耳畸形的发生可能存在关联,代谢组学为小耳畸形的研究提供了新的手段。

Objective Based on ear cartilage of congenital microtia,we tried to find the different metabolites and variational metabolic pathways on molecular level by gas chromatography time-of-flight mass spectrometry(GC-TOFMS).Methods Patients with microtia were collected from June 2017 to January 2018.During operation,the ear cartilage tissue was collected and labeled,and the case group was residual ear cartilage tissue of the patients with microtia,with 18 cases.The control group was normal ear cartilage tissue,total 18 cases.After the sample pretreatment,based on the technique of GC-TOFMS and the software of XploreMET,the differential metabolites in the residual ear cartilage of microtia were analyzed by orthogonal partial least square discriminant analysis(OPLS-DA)and its variable importance projection(VIP)and U test statistical method.Compared with the metabolite database,the metabolic pathway enrichment analysis(MPEA)was used to screen the most related metabolic pathways.Results According to the multidimensional statistical analysis,it is different in the metabolites of ear cartilage between the two groups.According to the verification of single dimensional statistical analysis,14 specific biomarkers were identified,and their P value was less than 0.05.According to the metabolic pathway enrichment analysis(MPEA),3 metabolic pathways had statistically significant difference,including arginine metabolism,taurine and the taurine metabolism,pantothenate and CoA metabolism,beta-alanine metabolism,glycerophospholipid metabolism,P<0.05.Conclusions Arginine,taurine,L-cysteine and pantothenic acid may have an association with microtia,which proved that it is feasible to study the pathogenesis of microtia by metabonomics.