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目的探讨蜘蛛毒素GsMTx-4对在中国仓鼠卵巢癌(CHO)细胞中表达的心脏力敏感大电导钾通道(SAKca通道)电流的影响。方法在CHO细胞中瞬时转染以表达SAKca通道蛋白,采用电极内倒灌流法和内面向外式膜片钳技术研究SAKca通道电流特性及GsMTx-4对SAKca通道电流的影响。结果当膜电位在-140~100mV范围时,SAKca通道电导值为(276.6±4.5)pS。与0mmHg压力相比,-30mmHg的压力使SAKca通道开放频率(Po)-膜电位曲线向左平移了(23.3±3.1)mV。通过倒灌流法在电极内加入100nmol/L的GsMTx-4,SAKca通道Po在10min内降低了75%左右。结论 GsMTx-4对SAKca通道电流有抑制作用。
Objective To investigate the effect of spider toxin GsMTx-4 on cardiac current-sensitive large conductance potassium channel (SAKca channel) currents expressed in Chinese hamster ovary cancer (CHO) cells. Methods Transient transfection of SAKca channel protein in CHO cells was performed. The effects of GsMTx-4 on the currents of SAKca channels were studied by the method of electrode-in-stream and patch-clamp technique. Results The conductivity of SAKca channel was (276.6 ± 4.5) pS when the membrane potential was in the range of -140 ~ 100mV. Compared to 0 mmHg pressure, a pressure of -30 mmHg translates the SAKca channel open-circuit (Po) -membrane potential curve to the left (23.3 ± 3.1) mV. By adding the flow-through method, 100 nmol / L GsMTx-4 was added to the electrode, and the SAKca channel Po decreased by 75% within 10 min. Conclusion GsMTx-4 has inhibitory effect on SAKca channel current.