伞形科辛味中药独活、蛇床子的主要成分及组合对大鼠离体胸主动脉的作用
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摘要
目的:研究伞形科辛味中药独活、蛇床子的3种主要成分蛇床子素、欧前胡素、异欧前胡素对大鼠离体胸主动脉舒张作用,探讨3种成分最佳配伍组合及其作用机制,探索成分组合与药效活性的相关性。方法:采用离体血管环实验,分别测定蛇床子素、欧前胡素、异欧前胡素3种成分舒张血管的量效曲线。采用均匀设计法得到3种成分不同组合与血管活性的关系,优化配比得到最优组合(组合药),并进行试验验证。观察组合药对大鼠胸主动脉张力的作用,记录KCl预收缩大鼠离体胸主动脉环的张力变化。采用左旋硝基精氨酸甲酯(L-NAME,3×10~(-4)mol/L)、左旋硝基精氨酸(L-NNA,3×10~(-4)mol/L)、ODQ(1×10~(-5)mol/L)、吲哚美辛(INDO,1×10~(-5)mol/L)、四乙基硝酸铵(TEA,1×10~(-5)mol/L)预孵后,观察组合药对血管张力变化的影响。并观察其对CaCl_2引起的外Ca~(2+)内流的影响。结果:蛇床子素、欧前胡素、异欧前胡素均对KCl预收缩离体胸主动脉环有舒张作用,并呈剂量依赖性。3种成分的最优组合为蛇床子素14.25μg/mL、欧前胡素148.97μg/mL、异欧前胡素202.12μg/mL。经药效验证,组合药对KCl预收缩血管环具有舒张作用,去除内皮后,该作用显著减弱(P<0.01)。预先用L-NAME、L-NNA、ODQ孵育后,组合药的舒张作用减弱,与未经抑制剂处理的对照组比较有统计学意义(P<0.05)。组合药对CaCl_2外Ca~(2+)内流所致最大收缩效应有显著抑制作用(P<0.01)。结论:蛇床子素、欧前胡素、异欧前胡素3种成分对大鼠离体胸主动脉的舒张作用具有协同增效之功,其可能的作用机制主要与抑制血管平滑肌上的Ca~(2+)通道,减少细胞外Ca~(2+)内流及抑制内皮细胞上NO途径有关。
Objective: To investigate the effects of osthole, imperatorin and isoimperatorin, which are the three main components of Radix Angelicae pubescens and Fructus Cnidii, on isolated thoracic aorta in Wistar rats, and explorethe optimal combination of the three components and the potential mechanism, and the correlation between combination and pharmacodynamic activity. Methods: The dose-effect curves of vasodilatation of osthole, imperatorin and isoimperatorin was measured by in vitro vascular ring experiment. The homogenous design method was used to obtain the relationship between the different combinations of three components and vascular activity, and the optimal combination(combination drug) was obtained by optimizing the ratio, and the experimental verification was conducted. The effect of the optional combination on vascular tension in rats was evaluated, and the effect of the optimal combination on endothelium-intact/denuded thoracicaortic rings pre-contracted with KCl was recorded. The effect of the optimal combination on endothelium-intactaortic rings, pre-inhibited separately with NG-nitro-LArginine methyl ester(L-NAME, 3×10~(-4) mol/L), NG-Nitro-L-arginine(L-NNA, 3×10~(-4) mol/L), ODQ(1×10~(-5) mol/L), indometacin(INDO, 1×10~(-5) mol/L) and tetraethyl ammoniumnitrate(TEA, 1×10~(-5) mol/L) was observed, and the effect of CaCl_2-induced Ca~(2+)influx was also observed. Results: Osthole, imperatorin and isoimperatorin all had a diastolic effect on KCl pre-contracted thoracic aortic rings in a dose-dependent manner. The optimum combination of the three components was 14.25μg/mL, 148.97μg/mL and 242.12μg/mL, respectively. The combination had a diastolic effect on the KC1 pre-contracted vascular rings. After removing the endothelium, the effect was significantly reduced(P<0.01). The diastolic effect of the combination was weakened after pre-incubation with L-NAME, L-NNA and ODQ, which was statistically significant compared with the control group without inhibitor treatment(P<0.05). The combination had a significant inhibitory effect on the maximum contraction induced by CaCl_2 external calcium influx(P<0.01). Conclusion: The diastolic effect on isolated thoracic aorta in rats of the combination of the three components of osthole, imperatorin and isoimperatorin was better than that of the sum of the three components alone, indicating that the combination of the three components had synergistic effect. The mechanism may be not only associated with inhibition of Ca~(2+) channels on the vascular smooth muscle, which leads to the decrease of Ca~(2+) influx, but also related to the NO pathway on the endothelial cells.
Objective: To investigate the effects of osthole, imperatorin and isoimperatorin, which are the three main components of Radix Angelicae pubescens and Fructus Cnidii, on isolated thoracic aorta in Wistar rats, and explorethe optimal combination of the three components and the potential mechanism, and the correlation between combination and pharmacodynamic activity. Methods: The dose-effect curves of vasodilatation of osthole, imperatorin and isoimperatorin was measured by in vitro vascular ring experiment. The homogenous design method was used to obtain the relationship between the different combinations of three components and vascular activity, and the optimal combination(combination drug) was obtained by optimizing the ratio, and the experimental verification was conducted. The effect of the optional combination on vascular tension in rats was evaluated, and the effect of the optimal combination on endothelium-intact/denuded thoracicaortic rings pre-contracted with KCl was recorded. The effect of the optimal combination on endothelium-intactaortic rings, pre-inhibited separately with NG-nitro-LArginine methyl ester(L-NAME, 3×10~(-4) mol/L), NG-Nitro-L-arginine(L-NNA, 3×10~(-4) mol/L), ODQ(1×10~(-5) mol/L), indometacin(INDO, 1×10~(-5) mol/L) and tetraethyl ammoniumnitrate(TEA, 1×10~(-5) mol/L) was observed, and the effect of CaCl_2-induced Ca~(2+)influx was also observed. Results: Osthole, imperatorin and isoimperatorin all had a diastolic effect on KCl pre-contracted thoracic aortic rings in a dose-dependent manner. The optimum combination of the three components was 14.25μg/mL, 148.97μg/mL and 242.12μg/mL, respectively. The combination had a diastolic effect on the KC1 pre-contracted vascular rings. After removing the endothelium, the effect was significantly reduced(P<0.01). The diastolic effect of the combination was weakened after pre-incubation with L-NAME, L-NNA and ODQ, which was statistically significant compared with the control group without inhibitor treatment(P<0.05). The combination had a significant inhibitory effect on the maximum contraction induced by CaCl_2 external calcium influx(P<0.01). Conclusion: The diastolic effect on isolated thoracic aorta in rats of the combination of the three components of osthole, imperatorin and isoimperatorin was better than that of the sum of the three components alone, indicating that the combination of the three components had synergistic effect. The mechanism may be not only associated with inhibition of Ca~(2+) channels on the vascular smooth muscle, which leads to the decrease of Ca~(2+) influx, but also related to the NO pathway on the endothelial cells.
引文
[1]苗明三,马霄,王灿.中药有效组分配伍研究的探讨.中药新药与临床药理,2009,20(5):487-490
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[3]李振坤,陈建新,杨洪军,等.基于LARS算法的川芎成分组合与血管活性关系分析.中国实验方剂学杂志,2009,15(3):24-27
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[5]李杨,张梅.中药组分配伍研究方法及应用实践.中药与临床,2016,7(5):51-53,64
[6]杨金果,李珩,李运伦.中药有效组分配伍的研究进展.上海中医药杂志,2012,46(3):89-92
[7]BAIK J S,SOHN J T,OK S H,et al.Levobupivacaine-induced contraction of isolated rat aorta is calcium dependent.Can J Physiol Pharmacol,2011,89(7):467-476
[8]SEOK Y M,KIM H Y,GARMAA O,et al.Effects of magnolol on vascular contraction in rat aortic rings.Clin Exp Pharmacol Physiol,2012,39(1):28-36
[9]张映桥,余涛,许激扬,等.天麻素对离体大鼠胸主动脉环的舒张作用及其机制.中国中药杂志,2012,37(14):2135-2138
[10]XIE Y L,ZHOU M,MA H H,et al.Vasorelaxation effects of gastrodin on isolated thoracic aorta rings in rats.Chin J Integr Med,2015,21(12):944-948
[11]翟向红.姜酚对家兔离体胸主动脉环的作用及机制.郑州:郑州大学,2013
[12]NELSON M T,QUAYLE J M.Physiological roles and properties of potassium channels in arterial smooth muscle.Am J Phsiol,1995,268(4):799-782
[2]盛华刚.中药组分配伍模式的研究进展.中国实验方剂学杂志,2013,19(16):349-352
[3]李振坤,陈建新,杨洪军,等.基于LARS算法的川芎成分组合与血管活性关系分析.中国实验方剂学杂志,2009,15(3):24-27
[4]王阶,郭丽丽,王永炎.中药方剂有效成(组)分配伍研究.中国中药杂志,2006,31(1):5-9
[5]李杨,张梅.中药组分配伍研究方法及应用实践.中药与临床,2016,7(5):51-53,64
[6]杨金果,李珩,李运伦.中药有效组分配伍的研究进展.上海中医药杂志,2012,46(3):89-92
[7]BAIK J S,SOHN J T,OK S H,et al.Levobupivacaine-induced contraction of isolated rat aorta is calcium dependent.Can J Physiol Pharmacol,2011,89(7):467-476
[8]SEOK Y M,KIM H Y,GARMAA O,et al.Effects of magnolol on vascular contraction in rat aortic rings.Clin Exp Pharmacol Physiol,2012,39(1):28-36
[9]张映桥,余涛,许激扬,等.天麻素对离体大鼠胸主动脉环的舒张作用及其机制.中国中药杂志,2012,37(14):2135-2138
[10]XIE Y L,ZHOU M,MA H H,et al.Vasorelaxation effects of gastrodin on isolated thoracic aorta rings in rats.Chin J Integr Med,2015,21(12):944-948
[11]翟向红.姜酚对家兔离体胸主动脉环的作用及机制.郑州:郑州大学,2013
[12]NELSON M T,QUAYLE J M.Physiological roles and properties of potassium channels in arterial smooth muscle.Am J Phsiol,1995,268(4):799-782