湖北省自然科学基金资助项目(SJ-971073).
This work was supported by a grant from The Natural Science Foundation of Hubei Provience (SJ-971073).
为研究蝎毒素BmkTXKβ对家兔心房肌细胞瞬间外向钾电流(Ito)的影响,采用全细胞膜片钳技术记录应用BmkTXKβ前后的Ito电流. 结果显示BmkTXKβ(1 μmol/L)使心房肌细胞Ito(刺激电压为+50 mV)从(13.63±0.87)pA/pF减少到(7.98±0.78)pA/pF,抑制率为41.4% (n=16, P<0.001).冲洗后, Ito部分恢复至(11.18±0.82)pA/pF (n=6, P<0.01,与给药后比较).在0.01~100 μmol/L范围内BmkTXKβ呈浓度依赖性地抑制Ito,IC50的均值为0.95 μmol/L (n=10, P<0.01),但无频率依赖性(n=6,P>0.05). 1 μmol/L的BmkTXKβ可使Ito通道的失活动力学曲线明显左移,V1/2分别为(-23.6±2.7) mV和(-35.3±3.6) mV(n=8,P<0.05),曲线斜率基本不变.同时可使Ito通道的恢复过程明显减慢,恢复曲线右移,τ值从(51.2±8.5) ms延长至(93.5±13.4) ms(n=9,P<0.01),但不影响其激活过程.据此推断BmkTXKβ对家兔心房肌细胞Ito具有显著的抑制作用,主要作用于失活过程,延长该通道的恢复时间.
In order to study the effects of BmkTXKβ on transient outward potassium current(Ito) of isolated rabbit atrial myocytes,standard whole-cell patch clamp technique was used to record Ito before and after administration of extracellular BmkTXKβ with multiple concentrations. The results showed that: (1) At a dose of 1 μmol/L, it decreased Ito by 41.4% (n=16, P<0.001) at membrane potential of +50 mV [from (13.63±0.87) pA/pF to (7.98±0.78) pA/pF]. After washout, Ito restored to (11.18±0.82) pA/pF (n=6, P<0.01). (2) It significantly reduced Ito in a clearly concentration-dependent manner at the range of 0.01~100μmol/L with an IC50 value of 0.95 μmol/L (n=10, P<0.01), but without any change in frequency-dependence (n=6,P>0.05). (3) In the absence and presence of BmkTXKβ (1 μmol/L),the activation curves from relative conductance almost overlapped, whereas steady-state inactivation curve shifted to left from (-23.6±2.7) mV to (-35.3±3.6) mV at V1/2 point significantly (n=8,P<0.05). The time for 50% recovery delayed obviously from(51.2±8.5) ms to (93.5±13.4) ms in the absence and presence of 1 μmol/L BmkTXKβ(n=9,P<0.01). The results show that BmkTXKβ exerts direct blocking effect on Ito in rabbit atrial myocytes, which is mainly caused by a strong suppression effect on inactivation duration and prolongation of the recovery duration from inavtivation.
胡丹,黄从新,江洪,李庚山,曹志贱,李文鑫,王世敏.蝎毒素BmkTXKβ对家兔心房肌细胞瞬间外向钾电流的抑制作用[J].生物化学与生物物理进展,2003,30(2):266-271
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