β-细辛醚对气道副交感节前运动神经元突触传递的抑制作用
摘要
细辛是我国的一种传统中药,临床应用已有几个世纪。文献报道,细辛会引起急性呼吸毒性作用,但相关的毒性成分、机制等都未明确。细辛经过煎煮后使用,其毒性便会大幅降低,故人们推测细辛挥发油可能是导致其呼吸毒性作用的主要物质。目前,人们已经分离了许多细辛挥发油的单体,包括α-和β-细辛醚(cis-1-Propenyl-2,4,5-trimethoxybenzene)、甲基丁香酚(]methyl eugenol)和黄樟醚(safrole)等。在最近的研究中,在麻醉兔用细辛散剂灌胃,观察到膈神经放电频率和幅度均被抑制,提示细辛的呼吸毒性可能与其对延髓呼吸中枢的抑制作用有关。
呼吸节律由延髓腹外侧区的前包钦格复合体(Pre-Botzinger complex, PBC)和邻近的一些神经元产生。这些区域的起博神经元通过兴奋性突触传递和电突触相互联系,并向颅神经前运动环路(cranial premotor circuit)以及延髓-脊髓吸气性前运动神经元(bulbospinal inspiratory premotor neuron)投射,延髓-脊髓吸气性前运动神经元再支配脊髓膈神经和肋间神经吸气运动神经元。有证据显示：呼吸节律发生器和气道副交感节前运动神经元(airway preganglionic parasympathetic motoneurons, APPMs)在解剖和功能上是相互“藕联”的。气道副交感神经或它们的节前细胞体与舌下神经和膈神经的吸气电活动同时相发生。支配膈神经前运动神经元的一组延髓呼吸神经元同时也支配APPMs。
离体延髓脑片是研究呼吸节律产生和气道副交感神经节前运动神经元的行之有效的模型。在我们的研究中,我们利用这个模型来研究细辛的有效成分p-细辛醚对中枢性呼吸驱动和对APPMs紧张性突触后活动的影响。
结果如下：
1.细辛乙醇提取物抑制了舌下神经根放电活动的频率和强度,说明细辛酯溶性成分有中枢呼吸毒性作用。
2.p-细辛醚浓度依赖性地减少舌下神经根吸气性放电活动的持续时间和面积。放电活动频率和幅度在最初不受影响或暂时增加,其后被进行性抑制。随着舌下神经根放电被抑制,APPMs的紧张性和相位性(phasic)突触后电流(包括兴奋性和抑制性)也进行性减弱。提示p-细辛醚对延髓呼吸神经元网络神经传递的抑制参与了细辛的呼吸毒性作用。
Therapeutic application of Asarum, a herbal medicine that has been used for centuries, reportedly causes acute respiratory disturbance. The responsible constituents, the sites of action, and the mechanisms involved in this side effect are unclear. It is generally believed, however, that components of Asarum volatile oils may be the cause of Asarum-induced respiratory disturbance, because the risk of this complication is reduced when Asarum is boiled prior to use. Researchers have isolated many of the Asarum volatile oil constituents, including α-and (3-asarone (cis-l-Propenyl-2,4,5-trimethoxybenzene), methyl eugenol, and safrole. In one recent study, intragastric application of Asarum pulvis inhibited the frequency and intensity of phrenic nerve discharges in anesthetized rabbits, suggesting the acute respiratory disturbance caused by Asarum may be a result of actions at the medullary respiratory centers.
Respiratory rhythm is generated in the Pre-Botzinger complex (PBC) and nearby in the ventrolateral medulla. Pacemaker neurons in this area are interconnected by excitatory synapses and gap junctions, and send projections to cranial premotor circuits and bulbospinal inspiratory excitatory neurons that project to spinal phrenic and intercostal inspiratory motoneurons. Some evidence has suggested that the respiratory rhythm generator and the airway preganglionic parasympathetic motoneurons (APPMs) may be coupled both anatomically and functionally. The airway parasympathetic nerves or their preganglionic cell bodies fire in phase with the hypoglossal and the phrenic nerves. A subset of the medullary respiratory neurons that project to the phrenic motoneurons also innervate the APPMs.
The brainstem slice is a well-established model to study the respiratory rhythm generator and the airway autonomic premotor neurons. In the present work, we utilized this model to investigate the effects of β-asarone, a volatile constituent of Asarum, on the central respiratory drive and the tonic postsynaptic activity of the APPMs.
Asarum ethanol extract inhibited the frequency and intensity of the hypoglossal respiratory-like bursts. β-Asarone caused progressive decreases in the duration and area of the hypoglossal bursts in a concentration-dependent manner. The frequency and amplitude of the bursts were initially unaltered or temporarily increased, but were then inhibited progressively after prolonged exposure. As with the inhibition of the hypoglossal bursts, the tonic and the phasic excitatory and inhibitory postsynaptic currents in the APPMs were attenuated. These data suggest that the Asarum-caused acute respiratory disturbance involves β-asarone-induced inhibition of neurotransmission in the medullary respiratory neuronal network.
呼吸节律由延髓腹外侧区的前包钦格复合体(Pre-Botzinger complex, PBC)和邻近的一些神经元产生。这些区域的起博神经元通过兴奋性突触传递和电突触相互联系,并向颅神经前运动环路(cranial premotor circuit)以及延髓-脊髓吸气性前运动神经元(bulbospinal inspiratory premotor neuron)投射,延髓-脊髓吸气性前运动神经元再支配脊髓膈神经和肋间神经吸气运动神经元。有证据显示：呼吸节律发生器和气道副交感节前运动神经元(airway preganglionic parasympathetic motoneurons, APPMs)在解剖和功能上是相互“藕联”的。气道副交感神经或它们的节前细胞体与舌下神经和膈神经的吸气电活动同时相发生。支配膈神经前运动神经元的一组延髓呼吸神经元同时也支配APPMs。
离体延髓脑片是研究呼吸节律产生和气道副交感神经节前运动神经元的行之有效的模型。在我们的研究中,我们利用这个模型来研究细辛的有效成分p-细辛醚对中枢性呼吸驱动和对APPMs紧张性突触后活动的影响。
结果如下：
1.细辛乙醇提取物抑制了舌下神经根放电活动的频率和强度,说明细辛酯溶性成分有中枢呼吸毒性作用。
2.p-细辛醚浓度依赖性地减少舌下神经根吸气性放电活动的持续时间和面积。放电活动频率和幅度在最初不受影响或暂时增加,其后被进行性抑制。随着舌下神经根放电被抑制,APPMs的紧张性和相位性(phasic)突触后电流(包括兴奋性和抑制性)也进行性减弱。提示p-细辛醚对延髓呼吸神经元网络神经传递的抑制参与了细辛的呼吸毒性作用。
Therapeutic application of Asarum, a herbal medicine that has been used for centuries, reportedly causes acute respiratory disturbance. The responsible constituents, the sites of action, and the mechanisms involved in this side effect are unclear. It is generally believed, however, that components of Asarum volatile oils may be the cause of Asarum-induced respiratory disturbance, because the risk of this complication is reduced when Asarum is boiled prior to use. Researchers have isolated many of the Asarum volatile oil constituents, including α-and (3-asarone (cis-l-Propenyl-2,4,5-trimethoxybenzene), methyl eugenol, and safrole. In one recent study, intragastric application of Asarum pulvis inhibited the frequency and intensity of phrenic nerve discharges in anesthetized rabbits, suggesting the acute respiratory disturbance caused by Asarum may be a result of actions at the medullary respiratory centers.
Respiratory rhythm is generated in the Pre-Botzinger complex (PBC) and nearby in the ventrolateral medulla. Pacemaker neurons in this area are interconnected by excitatory synapses and gap junctions, and send projections to cranial premotor circuits and bulbospinal inspiratory excitatory neurons that project to spinal phrenic and intercostal inspiratory motoneurons. Some evidence has suggested that the respiratory rhythm generator and the airway preganglionic parasympathetic motoneurons (APPMs) may be coupled both anatomically and functionally. The airway parasympathetic nerves or their preganglionic cell bodies fire in phase with the hypoglossal and the phrenic nerves. A subset of the medullary respiratory neurons that project to the phrenic motoneurons also innervate the APPMs.
The brainstem slice is a well-established model to study the respiratory rhythm generator and the airway autonomic premotor neurons. In the present work, we utilized this model to investigate the effects of β-asarone, a volatile constituent of Asarum, on the central respiratory drive and the tonic postsynaptic activity of the APPMs.
Asarum ethanol extract inhibited the frequency and intensity of the hypoglossal respiratory-like bursts. β-Asarone caused progressive decreases in the duration and area of the hypoglossal bursts in a concentration-dependent manner. The frequency and amplitude of the bursts were initially unaltered or temporarily increased, but were then inhibited progressively after prolonged exposure. As with the inhibition of the hypoglossal bursts, the tonic and the phasic excitatory and inhibitory postsynaptic currents in the APPMs were attenuated. These data suggest that the Asarum-caused acute respiratory disturbance involves β-asarone-induced inhibition of neurotransmission in the medullary respiratory neuronal network.
引文
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