年龄相关性听力损失大鼠耳蜗代谢组学的初步研究  

Study and Application of Cochlear Metabolomics in Rats with Age-related Hearing Loss

作　　者：万桓志 陈惠东 杨冰倩[1,2] 张园园 华清泉 Wan Huanzhi;Chen Huidong;Yang Bingqian;Zhang Yuanyuan;Hua Qingquan(Department of Otolaryngology-Head and Neck Surgery,Renmin Hospital of Wuhan University,Wuhan,430060,China;Research Institute of Otolaryngology-Head and Neck Surgery,Wuhan University)

机构地区：[1]武汉大学人民医院耳鼻咽喉头颈外科中心,武汉430060 [2]武汉大学耳鼻咽喉头颈外科研究所

出　　处：《听力学及言语疾病杂志》2024年第3期265-270,共6页Journal of Audiology and Speech Pathology

基　　金：国家专科疾病诊疗中心建设培育资金(ZX0000000037)。

摘　　要：目的应用大鼠耳蜗代谢组学技术探索年龄相关性听力损失的发生机制。方法选择2月龄(年轻组)和14月龄(老年组)SD大鼠各15只,通过听性脑干反应(ABR)检测各组大鼠听觉功能;利用HE染色观察两组耳蜗组织形态,并通过流式细胞术检测耳蜗组织氧化应激状态。每组选取5只采用非靶向超高效液相色谱-串联质谱(LC-MS/MS)技术对耳蜗组织进行代谢组学检测,分析衰老耳蜗代谢差异。结果相较于年轻大鼠,老年大鼠ABR检测8、16、32 kHz纯音及click声反应阈值显著升高(P<0.05),HE染色显示耳蜗衰老相关血管纹萎缩(P<0.05),流式细胞术检测提示氧化应激水平明显升高(P<0.05)。代谢组学检测老年组耳蜗中共鉴定出124种差异代谢物,其中包括鞘氨醇、全反式维生素甲酸、油酰胺在内的16种代谢物显著上调,而嘌呤、牛磺酸、硫胺素、脯氨酸及其衍生物等108种代谢物水平明显降低;提示蛋白质合成与分解、鞘脂代谢、嘌呤代谢、氧化应激相关信号转导、细胞死亡、辅酶生物合成等生理病理机制可能与耳蜗衰老有关。结论细胞衰老和耳蜗代谢功能失调可能是导致年龄相关性听力损失的重要机制。Objective Using cochlear metabolomics to study the mechanisms underlying age-related hearing loss in rat.Methods A total of 30 rats with 2-month-old(young group)and 14-month-old(old group)were selected,with 15 rats in each group.The auditory function in each group was detected by auditory brainstem response(ABR),the morphology of cochlear tissue in both groups was observed using HE staining,and the oxidative stress status of cochlear tissue was detected by flow cytometry.Five rats/groups were selected for metabolomic examination of cochlear tissue by untargeted ultra-high performance liquid chromatography-mass spectroscopy(LC-MS/MS)to analyze the metabolic differences in the aging cochlea.Results Compared with young group,ABR detection of tone burst at 8,16,and 32 kHz and click response thresholds were significantly higher in old group(P<0.05),HE staining showed cochlear senescence-related vascular stripe atrophy(P<0.05),and flow cytometric techniques suggested significantly higher levels of oxidative stress in old group(P<0.05).Metabolomics detection revealed that a total of 124 differential metabolites were identified in the cochlea of the old group,of which 16 metabolites including sphingosine,all-trans-retinoic acid,and oleamide were significantly upregulated,while the levels of 108 metabolites such as purine,taurine,thiamine,and proline and its derivatives were significantly decreased.The results suggested that physiopathological mechanisms such as protein synthesis and catabolism,sphingolipid metabolism,purine metabolism,oxidative stress-related signaling,cell death,and coenzyme biosynthesis may be involved in cochlear aging.Conclusion Cellular senescence and cochlear metabolic dysfunction may be important mechanisms of age-related hearing loss.

关 键 词：年龄相关性听力损失 代谢组学 耳蜗衰老 氧化应激 

分 类 号：R764.43[医药卫生—耳鼻咽喉科]