糖尿病性听力减退大鼠听觉神经通路相关蛋白质组学初步研究

Studies of Proteomics of the Auditory Neuropathway in Rats with Diabetes Related Hearing Impairment

目的探讨糖尿病相关性听力减退大鼠听觉神经通路蛋白质水平的变化。方法健康雄性Wistar大鼠30只随机分成对照组和实验组,每组15只,实验组按55 mg/kg一次性尾静脉注射链脲佐菌素制造糖尿病大鼠模型,对照组等量注射柠檬酸缓冲液,3个月后排除中耳炎,经ABR检测,选取正常听力大鼠及糖尿病性听力减退大鼠各10只,采用双向凝胶电泳分离技术(2-DE)、质谱技术(MS),对蜗核及听皮层组织进行蛋白质组差异分析,应用免疫组化技术验证已鉴定的差异蛋白质点。结果蛋白质组差异分析显示:正常大鼠和糖尿病性听力下降大鼠蜗核、听皮层组织部分蛋白质组间存在差异,经肽质指纹图谱(PMF)分析初步鉴定了Prohibitin、Synapsin、Cofilin-1、Annexin A3、T-complex protein 1、dihydropyrimidinase-related protein 2、Pyridoxal phosphate phosphatase等33个蛋白质(蜗核17个,听皮层16个),这些蛋白质可能与糖尿病性听力减退相关。通过对已鉴定蛋白质Prohibitin及Synapsin的免疫组化验证,证实这两种蛋白质在糖尿病性听力减退大鼠听皮层中表达下降。结论正常听力组和糖尿病性听力损失组大鼠蜗核、听皮层组织部分蛋白质组间存在差异,这些蛋白质可能与糖尿病性听力减退有关。

Objective To investigate the differential proteomics of the auditory neuropathway in rats with diabetes-related hearing impairment.Methods 30 healthy male Wistar rats were randomly divided into the control group and the experimental group,with 15 rats in each group.The experimental group was injected with streptozotocin 55 mg/kg once through the tail vein to create the diabetes rat model,and the control group was injected with citric acid buffer in the same amount.After three months,tympanitis was excluded.Tested by ABR,Two-dimensional gel electrophoresis(2-DE)and Mass spectrometry(MS)were used to detect the different proteins in the auditory cortex and the cochlear nuclei between the normal rats(10 rats)and the diabetic rats(10 rats).Immunohistochemistry was used to identify the different proteins.Results The differences of protein expression in the auditory cortex and the cochlear nuclei between the normal rats and the diabetes rats were found,by means of identification of protein spots from 2-DE gels.Peptide mass fingerprints(PMF)maps identified 33 proteins(7 in cochlear nucleas and 16 in auditory cortex),including Prohibitin,Synapsin,Cofilin-1,Annexin A3,T-complex protein 1,dihydropyrimidinase-related protein 2,Pyridoxal phosphate phosphatase,et al.The immunohistochemical study demonstrated that Prohibitin and Synapsin expression in the auditory cortex of the diabetes rats were decreased.Conclusion The discrepancy of the protein expression in the auditory cortex and the cochlear nuclei between the normal rats and the diabetes rats were found,which may be related to diabetes-related hearing impairment.