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目的用从豚鼠内耳组织中纯化的P0蛋白免疫大鼠,观察实验动物听神经髓鞘变化,推测P0蛋白在听神经病的发生发展中的作用。方法采用SDS-PAGE从内耳组织中分离、纯化P0蛋白,用Western Blot法鉴定纯化的P0蛋白。用纯化的P0蛋白作为抗原免疫大鼠,观察其听性脑干反应阈、耳声发射的变化、动物血清IgG水平以及听神经的髓鞘在光镜电镜下的形态学变化,免疫组织化学技术观察P0蛋白在听神经上的分布。结果免疫后实验组大鼠听力学显示听神经病的特征:耳声发射正常,而听性脑干反应严重异常;实验组血清IgG水平显著升高。电镜下可见听神经的脱髓鞘病变,免疫组织化学技术显示P0分布在蜗神经纤维的髓鞘上。结论P0蛋白作为抗原能诱发蜗神经的脱髓鞘病变,可能是听神经病的重要发病机制之一。
Objective To observe the changes of auditory myelin sheath in guinea pigs treated with P0 protein purified from the inner ear tissue of guinea pig and to investigate the role of P0 protein in the development of auditory neuropathy. Methods P0 protein was isolated and purified from inner ear tissue by SDS-PAGE, and the purified P0 protein was identified by Western Blot. The purified P0 protein was used as an antigen to immunize the rats to observe the changes of auditory brainstem response threshold, otoacoustic emissions, serum IgG level and morphology of auditory nerve myelin under light microscopy. Immunohistochemistry P0 protein in the auditory nerve distribution. Results After the immunization, the auditory neurology of the rats in experimental group showed the characteristics of auditory neuropathy: normal otoacoustic emissions and severe abnormalities of auditory brainstem response. Serum IgG levels in the experimental group were significantly increased. Electron microscope shows demyelinating lesions of the auditory nerve. Immunohistochemistry shows that P0 is located on the myelin of the cochlear nerve fibers. Conclusion P0 protein as an antigen can induce demyelinating lesions of the cochlear nerve, which may be one of the important pathogenesis of auditory neuropathy.