前庭神经鞘膜瘤外泌体对小鼠耳蜗基底膜的影响研究

Effects of Exosomes Derived from Vestibular Schwannoma on Cochlear Explants in Mise

目的研究前庭神经鞘膜瘤(vestibular schwannoma,VS)外泌体对小鼠耳蜗基底膜的影响,探讨其在VS患者听力下降中的潜在作用。方法收集2020年9月至2021年6月于上海交通大学医学院附属第九人民医院耳鼻咽喉头颈外科手术治疗的6例Ⅱ、Ⅲ期中小型VS患者的肿瘤组织及临床资料,根据AAO-HNS听力分级分为听力好组(Good Hearing,GH)和听力差组(Poor Hearing,PH);采用超速离心法提取VS原代细胞上清中的外泌体,通过Western-blot、透射电镜、纳米流式鉴定外泌体;免疫荧光染色技术探究VS外泌体对小鼠耳蜗基底膜毛细胞形态及数量的影响。结果在一定浓度下,PH组外泌体造成小鼠基底膜毛细胞数量减少,细胞形态异常,细胞排列紊乱;而GH组外泌体对毛细胞的形态及数量几乎无影响;当外泌体浓度下降,PH组外泌体仅造成毛细胞出现极少数量的丢失。结论VS外泌体具有不同程度的耳毒性,对小鼠耳蜗毛细胞的损伤具有浓度依赖性。

Objective To investigate the role of exosomes derived from vestibular schwannoma(VS)in hearing loss,and to provide a theoretical basis and experimental foundation for deafness prevention and treatment in VS.Methods Tumor tissues and clinical data were collected from 6 VS patients who underwent surgeries at the Ninth People's Hospital of Shanghai Jiao Tong University School of Medicine from September 2020 to June 2022.The patients were divided into a Good Hearing(GH)and a Poor Hearing(PH)group according to the AAO-HNS hearing classification.Exosomes were extracted from VS primary cell supernatants by ultracentrifugation and identified by Western-blot,transmission electron microscopy,and nanofluidics.Effects of VS exosomes on the morphology and number of surviving hair cells in murine cochlear explants were assessed by immunofluorescence staining.Results At certain concentrations,exposure to VS exosomes from PH patients resulted in a decrease in the number of hair cells,abnormal cell morphology and disordered cell arrangement,in contrast to exposure to VS exosomes from GH patients which had almost no effect on the morphology and number of surviving hair cells.These effects diminished as the concentration of VS exosomes from PH patients decreased.Conclusion VS exosomes can be ototoxic in a concentration-dependent manner,whose strength may correlate with the patient’s hearing level.