基于耳蜗核钙结合蛋白含量的变化探讨复聪片延缓老年C57BL/6J小鼠听力下降的实验研究  

An experimental study on the efficacy of Fucong Tablet in delaying hearing loss in aged C57BL/6J mice and its pharmacological mechanism via determining content changes of calcium-binding protein in cochlear nucleus

作　　者：向红 彭斌[2] XIANG Hong;PENG Bin(Graduate School of Hunan University of Chinese Medicine,Changsha,410208,China)

机构地区：[1]湖南中医药大学研究生院,长沙410208 [2]湖南中医药大学第二附属医院,长沙

出　　处：《中国中西医结合耳鼻咽喉科杂志》2024年第3期161-167,共7页Chinese Journal of Otorhinolaryngology in Integrative Medicine

摘　　要：目的观察复聪片对C57BL/6J小鼠耳蜗核(CN)钙结合蛋白含量变化的影响及其延缓老年小鼠听力下降的效果以探讨其相关机制。方法6月龄C57BL/6J小鼠24只,经听性脑干反应测听(ABR)检测后随机分为复聪片治疗组与老年空白组,分别喂养含复聪片的饲料或普通饲料6月。干预期结束时,另取1月龄C57BL/6J小鼠12只作为青年空白对照组。各组动物分别行ABR测定评价听力水平,然后处死物并取材。组织标本切片,分别行尼氏染色、免疫组化染色(IHC)比较观察三组小鼠CN中尼氏体、钙结合蛋白(PV、CR、CB)的表达水平差异。结果青年空白组小鼠平均听阈明显低于老年空白组,阈值差异均具有显著性统计学意义(P<0.01);老年空白组与复聪片治疗组小鼠均出现不同程度的听阈升高,老年空白组升高更为明显(P<0.05)。老年空白组CN神经元中尼氏体、CB、CR含量均较青年空白组明显下降,PV含量较青年空白组升高(P<0.01~0.05);复聪片治疗组CN神经元中尼氏体、CB、CR含量高于老年空白组,PV低于老年空白组,但仅尼氏体、CB含量差异有统计学意义(P<0.05)。结论老年C57BL/6J小鼠耳蜗核神经元表现CB、CR下降含量、PV含量升高并伴随以神经元损伤及听力下降。复聪片能有效延缓其听力下降,其作用机制可能与抑制钙结合蛋白D-28K(CB)的减少而延缓耳蜗核神经元衰退有关。Objective To investigate the possible pharmacodynamic mechanism of Fucong Tablet(FCT)in delaying hearing loss in aged C57BL/6J mice by determining content changes of calcium binding protein(CBP)in cochlear nucleus(CN).Methods Following auditory brain response(ABR)detection,24 C57BL/6J mice,aged 6-month,were randomly divided into two groups,i.e.FCT treatment group(FCTTG)and elderly blank control group(EBCG).The mice in FCTTG were fed with the feed mixed with FCT,while those in EBCG fed with ordinary feed,for 6 months respectively.By the end of feeding period,another 12 C57BL/6J mice aged one-month were selected as young blank control group(YBG),and ABR audiometry was carried out to determine their hearing levels among all these animals of 3 groups,followed by samples of CN tissue collected when animals put into death,with the tissue samples treated routinely.Then,the content of Nissl body(NB)and the expressive levels of calcium binding protein(PV,CR,CB)in CN were determined among all these animals in three groups by Nissl staining and IHC respectively.Results The averaged hearing threshold of mice in YBG was significantly lower than that of EBCG determined at the 6th month and the 12th month respectively,with significantly statistical differences between them(P<0.01).Animals in EBCG and FCTTG all showed elevation in hearing threshold at various degrees,with the hearing threshold of mice in EBCG elevated much obviously and with markedly statistical significance between them(P<0.05).The contents of nissl bodies,CB and CR in CN neurons of EBCG were significantly lower than those of YBG,while the content of PV was higher in EBCG than that of YBG,with statistical significances(P<0.05).However,the content of nissl bodies,CB and CR in CN neurons of FCTTG were obviously higher than that of EBCG,while PV was lower than that of EBCG,only with significantly statistica differences in the contents of nissl bodies and CB between these two groups(P<0.05).Conclusion Cochlear nucleus neurons in aged C57BL/6J mice have developed de

关 键 词：老年性耳聋 耳蜗核 复聪片 疗效 药效学机制 

分 类 号：R285.5[医药卫生—中药学]