中药甘松挥发油对HEK细胞Nav1.5电流电压及电导电压影响.docVIP

中药甘松挥发油对HEK细胞Nav1.5电流电压及电导电压影响 　 作者：葛郁芝，吴志婷，胡朗吉， Yeh Jay 【摘要】 　　目的应用膜片钳技术研究HEK细胞Nav1.5电流电压依赖性Na+电流失活与激活曲线, 甘松挥发油对电流-电压关系(I-V)，电导-电压关系(G-V)曲线的影响, 以及对快Na+通道失活(H-I)的影响。方法应用膜片钳技术研究HEK细胞Nav1.5电压依赖性Na+电流失活曲线, IV和HI曲线。结果正常自然依赖性失活曲线向超极化移位，移动速度10 min内是4 mV。甘松挥发油能降低I－V曲线Na电流的峰电位，但不能改变其Na电位（ENa）从内流向外流的反转电位的方向。GV曲线明显向超极化方面移动，Vg1/2 =(-59.8±4.22) mV是对照自然失活曲线；Vg1/2 =(-58.8±5.69) mV 是10 PPM甘松挥发油；Vg1/2 =(-68.0±4.36) mV是药物被洗脱后失活曲线（P＜0.01，n=6）。在除去自然移动值后10 PPM甘松挥发油的Na电位失活作用明显。甘松挥发油应用前, 应用10 PPM甘松挥发油以及药物洗脱后H－I曲线与自然失活曲线比较，校正自然失活因素后10 PPM甘松挥发油实际移动Vg1/2是(18.0±4.50) mV (Plt;0.01, n=5)，洗脱40 min后接近对照状态。结论甘松挥发油对IV，GV曲线有向超极化移位的影响。对快Na+通道有加速失活与抑制激活的作用, 然而电压依赖性Na失活是可逆性的。这一作用奠定了甘松挥发油抗心律失常的作用机制。 【关键词】 甘松挥发油； HEK细胞； Nav1.5； 膜片钳 　　Abstract：ObjectiveTo investigate the effects of traditional Chinese medicine volatile oil of Nardostachy Chinesis Batal (TCMVONCB) on the curves of current-voltage relationship(I-V)，conductance-voltage relationship (G-V) and fast Na channel inactivation (H-I) of voltage-dependent in Nav1.5 channels in human embryonic kidney (HEK) cells.MethodsThe whole-cell patch-clamp technique and HEK cells were employed to research the effects of TCMVONCB on the curves of I-V，G-V and H-I of voltage-dependent current in Nav1.5 channels.ResultsTCMVONCB reduced the peak of Na current in the I-V curve, but could change its Na (ENa) reversal potential direction from within flows to outside　G-V curve move to the hyper-polarization, Vg1/2 =(-59.8±4.22)mV was control curve; Vg1/2 =(-58.8 ± 5.69)mV was 10 PPM TCMVONCB curve; Vg1/2 =(-68.0 ± 4.36)mV was for wash out curve (Plt;0.01, n=6). 10 ppm TCMVONCB has significant effects on the current of Nav1.5 after removing the natural movement in the value, the Vg1/2 after 10 pmm TCMVONCB was Vg1/2 =18.04.5 mV (Plt;0.01, n=5)comparing the control, after 40 min of wash out, the value of Vg1/2 remove back and close to the state of control. ConclusionTCMVONCB has the effects of hyper-polarizing shift on the curves of I-V and G-V and Na channel to acc