维生素C与过氧化氢对老年豚鼠耳蜗外毛细胞大电导钙激活钾通道电流的影响

Effects of Vitamin C and Hydrogen Dioxide on the Electric Current of Large-conductance Calcium-activated Potassium Channels in Isolated Outer Hair Cells of Old Guinea Pigs' Cochlears

赖丹;黄非;覃纲;

1:泸州医学院附属医院耳鼻咽喉-头颈外科

2:宜宾市第一人民耳鼻咽喉科

摘要
目的研究过氧化氢(H2O2)及维生素C对老年豚鼠耳蜗外毛细胞大电导钙激活钾通道(large conductance Ca2+-activated potassium channels,BKCa channels)电流的影响及其机制。方法采用急性酶分离方法分离25只老年豚鼠耳蜗外毛细胞,以膜片钳全细胞记录方式观察BKCa通道电流(5只豚鼠);记录到稳定、正常的BKCa通道电流后,向新鲜分离贴壁外毛细胞的2ml浴槽的浴液中加入H2O2稀释液40μl,使浴液中H2O2浓度为4μmol/L,观察H2O2对BKCa通道电流的影响(5只豚鼠);再分组加入维生素C溶液10、20、40μl(各组5只豚鼠),使浴液中维生素C终浓度分别为25、50、100μg/ml,观察H2O2和维生素C联合作用对BKCa通道电流的影响。结果 1膜片钳全细胞记录模式下,记录到一串幅值较大、快速激活、几乎不失活的电流,激活电压大于-40-30mV,电流随膜电位的增加而增强,并表现出外向整流的特性;加入BKCa通道特异性阻断剂伊比利亚毒素(iberiotoxin,IbTX)100nmol/L后,通道活动完全阻断,证实为BKCa通道电流。2加入H2O2后,用药3分钟内,当VT为+50mV时,BKCa通道峰值电流密度最大值从22.09±0.27pA/pF升至43.53±1.09pA/pF,增幅97.06%。H2O24μmol/L+25、50、100μg/ml维生素C时,BKCa通道电流表现浓度依赖性抑制,电流幅值和峰值电流密度随着维生素C浓度增加而减小,I-V曲线下降,洗脱后仍不能恢复至加药前正常水平。结论老年豚鼠耳蜗外毛细胞存在氧自由基/BKCa途径,而维生素C可减轻氧自由基对外毛细胞BKCa通道的影响。
关键词
大电导钙激活钾通道;外毛细胞;氧自由基;维生素C;过氧化氢
基金项目(Foundation):
四川省卫生厅科学研究项目(130358)资助
作者
赖丹;黄非;覃纲;
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