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目的:探讨耳聋左慈丸及其有效拆方(熟地黄泽泻药对、泽泻单味药)对庆大霉素诱导内耳听毛细胞凋亡相关蛋白及酶活性变化的影响。方法:复制内耳螺旋器体外培养庆大霉素损伤模型,采用原位荧光标记及免疫荧光标记观察庆大霉素损伤毛细胞区域凋亡酶Caspases活性、凋亡相关蛋白Bcl-2和Bax蛋白表达量的改变及中药的影响。结果:与正常对照组比较,0.3mmol.L-1庆大霉素能够引起内耳毛细胞Caspase-3,Caspase-9酶活性明显增强,凋亡调节因子Bcl-2/Bax蛋白表达明显下降。与模型组比较,耳聋左慈丸对Caspase-9酶活性有显著抑制作用,对Caspase-3酶活性有抑制趋势,对Bcl-2/Bax蛋白表达有明显上调作用;熟地黄泽泻药对、泽泻单味均能明显抑制Caspase-3,Caspase-9酶活性,显著上调Bcl-2/Bax蛋白表达。结论:耳聋左慈丸、熟地黄泽泻药对及泽泻单味可能通过纠正线粒体凋亡信号转导途径的异常变化,拮抗庆大霉素对毛细胞的损伤。
Objective: To investigate the effects of Deqin Zuo Ci Pills and its effective decomposed formulas (Rehmannia melanoides laxative and Alisma orientale) on changes of apoptosis-related proteins and enzyme activities induced by gentamicin in inner ear hair cells. METHODS: The gentamycin injury model was cultured in vitro by the inner ear screw, and the activity of apoptosis enzyme Caspases and the expression of apoptosis-related proteins Bcl-2 and Bax were detected by in situ fluorescence and immunofluorescence. Changes in quantity and influence of Chinese medicine. RESULTS: Compared with the normal control group, 0.3 mmol.L-1 gentamicin could significantly increase the activity of Caspase-3 and Caspase-9 in hair cells of the inner ear, and the expression of Bcl-2/Bax protein decreased significantly. Compared with the model group, Erqi Zuoci Pill had a significant inhibitory effect on Caspase-9 enzyme activity, inhibited the activity of Caspase-3 enzyme, and significantly increased the expression of Bcl-2/Bax protein; Both diarrhea and single odor significantly inhibited the activity of Caspase-3 and Caspase-9, and significantly up-regulated the expression of Bcl-2/Bax protein. Conclusion: The deafness Zuo Ci Pill, Rehmannia melanoides laxative pair and the Alisma of Alisma orientalis may correct the abnormal changes of mitochondrial apoptosis signal transduction pathway and antagonize the damage of gentamycin to hair cells.