三种检测耳蜗毛细胞死亡模式方法的比较

Comparison of three methods for detecting the death modes of hair cells in the cochlea

目的评估三种检测耳蜗毛细胞死亡模式方法的优缺点。方法以噪声暴露诱导灰鼠耳蜗毛细胞死亡,采用DNA荧光染料碘化丙锭(Propidiumiodide,PI)染色法,原位末端标记法(TUNEL)和Caspase-3染色法标记毛细胞死亡模式,对比三种检测方法。结果PI染色法可将正常、凋亡、坏死和缺失的毛细胞区分开来,TUNEL法能初步鉴别毛细胞死亡模式,Caspase-3标记可定性凋亡和坏死的毛细胞。结论PI染色法快速、简便、可靠、经济,可用于定量检测凋亡和坏死的毛细胞。

Objective To explore the virtues and defects of three different methods for detecting the death modes of hair cell in the cochlea after noise trauma. Methods Chinchillas were exposed to a 4 kHz narrow band noise at 108 dB SPL for 1h. After the noise exposure, the organs of Corti were processed for detecting of hair cell death modes. The apoptotic and necrotic hair cells were distinguished by three methods including staining with propidium iodide (PI), TUNEL assay and caspase-3 labeling respectively. The specimens were examined under a fluorescence microscope and three methods were compared with each other. Results Normal, apoptotic, necrotic and missing hair cells could be distinguished with PI staining. The death modes of hair cell could be primarily differentiated by TUNEL assay. Caspase-3 labeling was a qualitative method for analyzing apoptotic and necrotic hair cells. Conclusion PI staining is a fast, simple, reliable and cheap method. It can be used to quantitatively analyze the death modes of hair cell in the cochlea after noise trauma.