复方健耳剂对C57BL/6J小鼠AHL毛细胞的保护作用

Protective effects of the Compound Healthy Ear Agent against presbycusis in C57BL/6J mice

目的探讨老年性耳蜗毛细胞损害与中药复方健耳剂两种喂药方法干预的作用。方法选择1月龄C57BL/6J小鼠22只用于本实验,其中4只小鼠每日饮用自来水直到出生后3个月作为幼龄对照组;6只小鼠每日饮用自来水直到出生后7个月作为老年性聋对照组;6只小鼠每日自动饮用中药复方健耳剂直到出生后7个月;另6只小鼠每日自动饮用同样中药至4个月后改用每日人工灌服直到出生后7个月。各组动物实验到期终止后,取耳蜗进行全耳蜗基底膜铺片,将全耳蜗内、外毛细胞计数结果输入计算机并应用耳蜗图软件进行耳蜗毛细胞密度对比分析,其中选择基底膜上重要的病变区间的毛细胞密度进行统计学分析。结果 3月龄对照组小鼠耳蜗外、内毛细胞缺损仅仅出现在耳蜗底回钩端区域;7月龄对照组外、内毛细胞缺损从底回基底膜起始端扩展到距离耳蜗顶端约40%区域;7月龄中药灌服组和自动饮用组动物的内、外毛细胞缺损范围和程度相似,均显著比7月龄对照组为轻(P<0.001)。结论中药复方健耳剂能够有效延缓C57BL/6J小鼠老年性耳蜗毛细胞损害的发生和发展,两种喂药方式所起作用相同(P>0.05),其药理机制可能与其改善微循环,清除活性氧,保护线粒体等作用相关。

Objective To investigate the efficacy of the Compound Healthy Ear Agent（CHEA）, a traditional Chinese medicine compound product, on age-related cochlear hair cell damage by two different administration methods.Methods Twenty two 1 month old C57BL/6J mice were divided into 4 groups to drink tap water until 3 months of age（young control group, n=4）, drink tap water until 7 months of age（presbycusis control group, n=6）, voluntarily drink CHEA until 7 months of age（n=6）, or voluntarily drink CHEA until 4 months of age, followed by CHEA feeing until 7months of age（n=6）. At the end of the experiment, the cochlear basilar membrane was micro-dissected out, trimmed and mounted in glycerin on glass slides as flat surface preparations. The number of cochlear hair cells along the entire length of cochlea was counted for cochleograms. The hair cells density at two different locations of interval lesions was statistically analyzed and compared among the four groups. Results Cochlear hair cells in the young control group showed missing cells only at the end of the basal turn. Hair cell missing in the presbycusis control group extended from the basal turn to about 40% from the apex. In both CHEA treated groups, the extent and degree of missing hair cells were less compared to the presbycusis control group（P〈0.001）. Conclusion The traditional Chinese medicine compound product CHEA appears to mitigate the development age-related cochlear hair cells damage, with similar results for both administration methods tested in this study. The mechanisms may involve improving microcirculation, eliminating reactive oxygen species and protecting mitochondria.