目的观察单独及联合使用丹酚酸B和延胡索乙素对大鼠心室肌细胞L-型钙通道的影响。方法采用急性酶解分离法获得大鼠的单个心肌细胞,使用全细胞膜片钳技术记录钙通道电流。观察给药前后钙通道电流峰值(钙电流活化后峰值点与完全失活后电流轨迹的垂直距离)的变化。结果单独使用丹酚酸B(1,10,100μmol/L)对钙电流峰值的抑制率分别为:(25.3±16.4)%(n=4),(44.6±24.0)%(n=6),(86.0±20.4)%(n=4)。单独使用延胡索乙素(10,30,100μmol/L)对钙电流峰值的抑制率分别为:(22.2±6.4)%(n=5),(27.4±1.6)%(n=3),(51.0±23.0)%(n=9);联合使用丹酚酸B(1μmol/L)和延胡索乙素(10μmol/L)对钙通道电流峰值的抑制作用强于单独使用丹酚酸B(1μmol/L)或延胡索乙素(10μmol/L),差异有统计学意义(P〈0.05);盐酸阿托品(14mmol/L)能够逆转延胡索乙素对L-型钙通道的抑制作用,而加强丹酚酸B的抑制作用。结论丹酚酸B和延胡索乙素对大鼠心室肌细胞L-型钙通道均有抑制作用,两者之间能够产生协同效应,并且这两种有效成分调控L-型钙通道的作用机制不同。
Objective To observe the effects of the separate and joint use of salvianolic acid B(SalB) and tetrahydropalmatine(THP) on the L-type calcium channel of rat ventricular myocytes. Methods Single isolated ventricular myocytes of rats were obtained using acute enzymolysis separation. The current of the L-type calcium channel was recorded using whole-cell patch clamp technique. Changes of the current peak value of the calcium channel(the vertical distance between the peak value point after activation of the calcium electric current and the electric current track after complete inactivation) were observed before and after medication. Results The inhibition rate of using SalB(at the dose of 1,10,and 100 μmol/L) alone on the current peak value of the calcium channel was respectively(25.3±16.4) %(n=4) ,(44.6±24.0) %(n=6) ,and(86.0±20.4) %(n=4) . That of using THP(at the dose of 10,30,and 100 μmol/L) alone on the current peak value of the calcium channel was respectively(22.2±6.4) %(n=5) ,(27.4±1.6) %(n=3) ,and(51.0±23.0) %(n=9) . The inhibition potency of joint use of SalB(1 μmol/L) and THP(10 μmol/L) on the current peak value of the calcium channel was stronger than using SalB(1 μmol/L) alone or THP(10 μmol/L) alone,showing statistical difference(P0.05) . Atropine hydrochloric acid(14 mmol/L) could reverse the inhibition of THP on the L-type calcium channel,while strengthening the inhibition of SalB. Conclusions Both SalB and THP showed inhibition on the L-type calcium channel of rat ventricular myocytes. They could generate synergistic effects. Besides,their action mechanisms for regulating the L-type calcium channel were different.