摘要
目的:在成功建立了过表达CXCR4骨髓间充质干细胞系(CXCR4-BMSCs)的基础上,观察CXCR4-BMSCs在庆大霉素致感音神经性聋的豚鼠耳蜗内的生存和迁移能力,探讨SDF-1/CXCR4轴是否能够介导BMSCs向耳蜗归巢。方法:分离并提取豚鼠骨髓间充质干细胞(BMSCs),并成脂肪、成骨、成神经诱导,鉴定其多向分化能力,建立CXCR4-BMSCs;取健康豚鼠50只,庆大霉素[100mg/(kg·d)]腹腔注射建立感音神经性聋动物模型,将成功致聋动物随机分为3个组:A组:感应神经性聋组(14只),耳蜗鼓阶途经植入灭菌注射用水;B组:感音神经性聋植入BMSCs(14只),耳蜗鼓阶途经植入;C组:感应神经性聋植入过表达CXCR4BMSCs组(14只),耳蜗鼓阶途径植入。并于2周、4周后进行听性脑干反应(ABR)检测,耳蜗组织冷冻切片荧光追踪植入细胞在耳蜗内的定向归巢情况。结果:耳聋动物造模成功后ABR检测3组听阈分别为(91.3±5.2)dB(SPL)、(90.7±4.8)dB(SPL)、(90.9±5.6)dB(SPL),差异无统计学意义(P>0.05)。A组豚鼠2周及4周听阈分别为(89.7±6.4)dB(SPL)和(89.2±6.7)dB(SPL),未见明显提高(P>0.05);B组豚鼠2周后听阈为(88.6±5.3)dB(SPL),差异无统计学意义(P>0.05),4周后听阈为(78.4±7.3)dB(SPL),差异有统计学意义(P<0.05);C组术后2周听阈为(75.3±7.8)dB(SPL),听力提高约15dB;4周后听阈为(62.1±8.4)dB(SPL),听力提高约28dB,均差异有统计学意义(P<0.05)。荧光追踪显示CXCR4-BMSCs定向归巢的细胞数量明显增多,且呈现出队列样排列。结论:SDF-1/CXCR4轴在BMSCs向耳蜗定向归巢及分化过程中发挥了重要作用,可提高庆大霉素致感音神经聋豚鼠的听力修复能力。
Objective：To observe the survival and migration ability of CXCR4-BMSCs in guinea pig cochlea with gentamycin induced sensorineural hearing loss,and to explore whether SDF-1/CXCR4 axis can mediate bone marrow mesenchymal stem cells（BMSCs）to cochlear homing.Method：BMSCs were isolated and extracted from guinea pigs.A bone marrow mesenchymal stem cell line with overexpression of CXCR4 was established.The animal model of sensorineural deafness was established by intraperitoneal injection of gentamicin [100 mg/（kg·d）]in 50 healthy guinea pigs.The successful deafness animals were randomly divided into three groups（14 rats in each group）：sterilizing water group,BMSCs transplantation group and CXCR4-BMSCs transplantation group.Auditory brainstem response（ABR）were performed at 2 weeks and 4 weeks respectively after the tansplantation through cochlear tympanic pathway.The directional homing of the implanted cells in the cochlea was traced by the frozen section fluorescence of the cochlear tissue.Result：The hearing thresholds of the three groups were（91.3±5.2）,（90.7±4.8）and（90.9±5.6）dB（SPL）respectively.There was no significant difference among them（P〈0.05）.In sterile Water group,the hearing thresholds were（89.7±6.4）dB and（89.2±6.7）dB respectively when detected on 2 and 4 weeks after transplantation.There were no significant difference（P〈0.05）;In BMSCs group,the hearing thresholds were（88.6±5.3）dB（SPL）on 2 weeks and（78.4±7.3）dB（SPL）on 4 weeks after transplantation.In CXCR4-BMSCs group,the hearing thresholds were（75.3±7.8）dB（SPL）on 2 weeks and（62.1±8.4）dB（SPL）on 4 weeks after transplantation.The differences were statistically significant（PThe hearing threshold was（75.3±7.8）dB（SPL）at 2 weeks after operation.The hearing enhancement was about 15 dB,the hearing threshold was（62.1±8.4）dB（SPL）at 4 weeks,the difference was statistically significant（P〈0.05）.Fluorescence tracing showed that the number of CXCR4-BMSCs homing cells was significantly increased,and showed a cohort like arrangement.Conclusion：The SDF-1/CXCR4 axis plays an important role in the directional homing and differentiation of the cells into the cochlea,which can improve the hearing repair ability of guinea pigs induced by gentamicin induced sensorineural deafness.
作者
黄少鹏
王晓燕
方超
吴丽华
林碧玉
叶青
HUANG Shaopeng1 WANG Xiaoyan2 FANG Chao3 WANG Xiaoyan1 WU Lihua1 LIN Biyu1 YE Qing1(1Department of Otorhinolaryngology, Provincial Clinical College, Fujian Medical University, Fujian Provincial Hospital, Fuzhou, 350001, China 2Department of Otorhinolaryngology, H＇os＂ pital of Northwesterm Polytechnical University,3 Department of Otorhinolaryngology, the First Hospital of Putia)
出处
《临床耳鼻咽喉头颈外科杂志》
CAS
北大核心
2018年第5期355-359,364,共6页
Journal of Clinical Otorhinolaryngology Head And Neck Surgery
基金
福建省科技厅自然科学基金面上项目资助(No：2014J01290)
福建省临床重点专科建设项目[No：闽卫医政(2015)593号]
关键词
干细胞
聋
感音神经性
耳蜗移植
CXCR4
stem cell
hearing loss, sensorineural
cochlear tansplantation
CXCR4
