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目的:研究附子含药血清对大鼠心肌细胞L型钙通道的影响,探讨附子抗缓慢型心律失常的作用机制。方法:附子水煎后水浴浓缩成1∶1,给予大鼠灌胃,每次15.6 g·kg-1,7 d后制备含药血清;细胞密度20个/m L,药物血清体积分数为10%。作用时间:从加药物血清20 min后开始记录,到1 h逐渐结束,各组样本数均为8;采用急性酶解法获得大鼠的心室肌细胞,用全细胞膜片钳技术记录L型钙通道电流,观察附子含药血清对单个心室肌细胞L型钙通道电流(Ica-t)幅度及动力学特性的影响。结果:10%空白血清组和10%附子含药血清组给药后的平均峰电流密度和平均激活率分别为:(-3.59±1.06)p A/p F(单位细胞膜面积的电流大小),12.87%(n=8,P<0.05)和(-4.01±0.88)p A/p F,51.17%(n=8,P<0.01);附子含药血清激活曲线用药前和用药后的V1/2和K(稳态半数激活电压和斜率因子)分别为(-12.66±0.44)m V和(5.70±0.35),(-14.71±0.88)m V和(5.68±0.72),(P<0.05,n=8);附子含药血清失活曲线用药前和用药后的V1/2和K分别为(-28.10±0.20)m V和(6.31±0.18),(-37.55±0.11)m V和(5.94±0.09),(P<0.05,n=8);附子含药血清恢复曲线用药前和用药后的τ为(171.01±11.13)ms和(160.15±4.97)ms(n=8),差异无统计学意义;附子含药血清可增加L型钙通道电流密度、激活曲线左移,失活曲线右移,对恢复曲线没有影响。结论:附子含药血清能增加心室肌细胞钙通道电流。
Objective: To study the effect of aconite-containing serum on L-type calcium channel in rat cardiomyocytes and to explore the mechanism of aconite-induced anti-arrhythmia. Methods: The aconite decoction was concentrated to 1: 1 in a water bath, and then intragastric administration was given to rats. The drug-containing serum was prepared after 15.6 g · kg-1 and 7 days. The cell density was 20 cells / m L and the drug serum volume fraction was 10 %. Duration of action: The drug was added from the serum 20 min after the start of recording to 1 h gradually ended, the number of samples in each group were 8; acute myocardial enzymolysis obtained rat ventricular myocytes, whole-cell patch clamp recording of L-type calcium channel The effects of aconite-containing serum on the amplitude and kinetics of L-type calcium channel current (Ica-t) in single ventricular myocytes were observed. Results: The average peak current density and average activation rate after administration of 10% blank serum group and 10% Fuzi drug serum group were: (- 3.59 ± 1.06) p A / p F (current per unit cell membrane area) 12.87% (n = 8, P <0.05) and (-4.01 ± 0.88) p A / p F, 51.17% (n = 8, (-12.66 ± 0.44) mV and (5.70 ± 0.35), (-14.71 ± 0.88) mV and (5.68 ± 0.72), respectively (P <0.05, n = 8). The inactivated serum of aconite-containing serum before and after treatment was (-28.10 ± 0.20) mV and (6.31 ± 0.18), (-37.55 ± 0.11) mV and (P <0.05, n = 8). The recovery curves of Fuzi containing serum before and after treatment were (171.01 ± 11.13) ms and (160.15 ± 4.97) ms (n = 8) Statistical significance; Fuzi serum containing L-type calcium channel can increase the current density, activation curve to the left, inactivation curve to the right, no effect on the recovery curve. Conclusion: Fuzi serum can increase the calcium current in ventricular myocytes.