摘要
目的观察纯中药制剂对庆大霉素致聋动物和耳蜗毛细胞的修复再生作用。方法选用73只健康青年豚鼠,行听力学检查后,53只动物连续肌肉注射庆大霉素(80mg.kg-1.d-1)24～30天致聋(GM组),其间死亡8只,剩余45只随机分成中药治疗组(25只)和致聋后对照组(20只),中药治疗组给予纯中药制剂"复聪汤Ⅰ号"口服液和"复聪汤Ⅱ号"滴耳液治疗,致聋后对照组同法给予生理盐水。另外20只豚鼠作为正常对照组,每天肌肉注射等量生理盐水。在治疗30、60和90天后对GM组两组动物分别行ABR、DPOAE检测,同时,每一时间点处死6只动物,左侧耳蜗行扫描电镜观察,右侧耳蜗铺片行光镜观察和毛细胞计数。结果实验前所有豚鼠的听功能正常;连续注射庆大霉素30天后,GM组豚鼠ABR反应阈值上升到50～80dB SPL,DPOAE幅值降低,光镜和电镜下表现为耳蜗毛细胞纤毛断碎、倒伏、融合和消失,多处毛细胞表皮板肿胀、突起和疱疹样变性,或毛细胞被完全挤出网状板,基底回和第三回耳蜗毛细胞严重消失;中药治疗30天后,80%的耳聋豚鼠的ABR反应阈恢复到30～50dB SPL,DPOAE幅值明显提高;耳蜗铺片和电镜观察显示耳蜗毛细胞数目有一定恢复,组织结构有修复现象,扫描电镜可见再生毛细胞仅出现少量纤毛,而致聋后对照组未发现此种现象;治疗60天后,耳蜗毛细胞数目明显增多,Corti器的毛细胞区有大量的增殖细胞出现,扫描电镜可见成小束的新生纤毛出现在毛细胞缺失部位,同时支持细胞大量增殖;治疗90天后,再生的静纤毛束已基本形成,尤其是耳蜗基底部位的毛细胞明显增多,听功能基本正常。结论纯中药制剂复聪汤对庆大霉素致聋豚鼠耳蜗毛细胞有一定的修复和再生作用。
Objective Damaged cochlea hair cells are the leading cause of permanent sensorineural deafness.This paper discusses the study on the effectiveness of Traditional Chinese Medicines(TCM) in treating ototoxicity associated with aminoglycoside antibiotics.Methods 73 adult guinea pigs were used in this experiment.Auditory brainstem response(ABR) and distortion product otoacoustic emission(DPOAE) were performed on the individual animals which were divided into three groups: 20 gentamicin(GM) damaged control group,25 GM treated group.The two group animals were intrperitoneally injected with GM 80 mg/kg per day for consecutive 28 d.20 guinea pigs were used as the control group.25 animals by GM-induced deafness were treated with Chinese Traditional Medicine of Fucong mixture liquid for consecutive 30 to 90 d while the same volume of physiological saline was given to GM-induced deafness group and normal control group at the same time.At 30,60 and 90 d,the experiment animals received ABR and DPOAE for the auditory functions in each group.Then 6 experiment animals were sacrificed after the auditory function assessments.Then the cochlear surfaces were prepared and the numbers of cochlea outer hair cells were counted under the light microscope and the morphological changes of cochlea were observed under the scanning electronic microscope(SEM).To GM-induced deafness group and normal control group in the same time.At the tearting 30,60 and 90d time,the experment animals ABR and DPOAE was assess the variation of auditory function in each group.In this time,6 experment animals was killing after auditory function assess,Cochlear surface preparation,the number of cochlea outer hair cells was counted under the light microscope and the morphological changes of cochlea were observed under the scanning electronic microscope(SEM).Results Before the experiment,all the animals had normal ABR thresholds and DPOAE amplitudes.After the GMs were injected 28d afterwards,the ABR thresholds increased to 50~80 dB SPL and the DPOAE amplitudes decreased significantly in the GM-inducted deafness group,especially in the high-frequency areas that correlated with the greatest hair cells loss to the base of cochlea.Light and SEM show that ototoxicity had damaged the hair cells from fragmentation to the swelling of the apical surface of the hair cells.At the 30d after the TCM treatment,there was a partial recovery of the ABR and DPOAE functions in that the cochlear hair cell loss decreased in the base turn,especially in the OHC missing region where numerous micovilli were noted on the developmental cuticular plate.After 60 d of TCM treatment,the stereocilia appeared on the developmental cuticular,and the ABR and DPOAE functions significantly recovered.At the 90d,the small bundle and small apical surfaces were observed on the developmental cuticular.In addition,some new hair cells appear with the intermediate morphology between hair cells and supporting cells.The microvilli were similar to the hair bundles.Some hair cells appear matured with stereocilia covering the entire apical surface and splaying the graded heights.Conclusion This experiment suggests that the TCM treatment has a certain curative effect on the cochlea hair cells of the subjects.TCM treatment can be promoted as a way for the new hair cells regeneration and recovery from ototoxic exposure.
出处
《听力学及言语疾病杂志》
CAS
CSCD
北大核心
2012年第5期471-479,共9页
Journal of Audiology and Speech Pathology
基金
国家自然科学基金(No
81170922)和横向研究课题资助
关键词
中药
耳蜗
毛细胞
再生
豚鼠
Traditional Chinese medicines(TCM)
Cochlear
Hair cell
Regeneration
Guinea pig
