Regulation of calcitonin gene-related peptide secretion from trigeminal nerve cells by botulinum toxin type A: Implications for migraine therapy
Regulation of calcitonin gene-related peptide secretion from trigeminal nerve cells by botulinum toxin type A: Implications for migraine therapy
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DOI：
10.1111/j.1526-4610.2004.04007.x
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发表时间：
2004-01-01
期刊：
影响因子：
5
通讯作者：
Cady, R
中科院分区：
文献类型：
--
作者：
Durham, PL;Cady, R;Cady, R
客观。-确定A型肉毒毒素对培养的三叉神经节神经元分泌降钙素基因相关肽的影响。肉毒杆菌毒素通过阻断神经肌肉接头处的乙酰胆碱释放而引起肌肉麻痹的能力是众所周知的。先前的研究和临床观察未能证明与肉毒杆菌毒素或肉毒杆菌中毒疾病有关的感觉变化。然而，最近的研究表明，A型肉毒杆菌毒素注射到颅周肌肉可能有预防偏头痛的好处。这一观察结果重新辩论的机制，由肉毒杆菌毒素A型介导的感觉抑制。原代培养的大鼠三叉神经节被用来确定是否A型肉毒杆菌毒素可以直接减少降钙素基因相关肽，神经肽参与偏头痛的潜在病理生理释放。用临床有效剂量的A型肉毒杆菌毒素或对照载体处理未处理的培养物或用去极化刺激物（氯化钾）或辣椒素（一种已知激活感觉C纤维的试剂）刺激的培养物3、6或24小时。使用特异性放射免疫测定法测定各种处理后分泌到培养基中的降钙素基因相关肽的量。高百分比（大于90%）的三叉神经节神经元中存在的1至3天的文化被证明表达降钙素基因相关肽。用去极化刺激（氯化钾）、炎性物质混合物或辣椒素治疗引起三叉神经元释放的降钙素基因相关肽显著增加（4- 5倍）。有趣的是，用治疗浓度的A型肉毒杆菌毒素（1.6或3.1单位）过夜处理三叉神经节培养物并不影响从这些神经元释放的降钙素基因相关肽的量。降钙素基因相关肽的刺激释放后，化学去极化与氯化钾或激活与辣椒素，然而，大大抑制了肉毒杆菌毒素，但不是由控制车辆。用1.6和3.1单位的A型肉毒杆菌毒素过夜处理观察到类似的抑制作用。A型肉毒杆菌毒素的这些浓度完全在局部注射容易达到的组织浓度范围内或以下。与对照值相比，将培养物与毒素孵育24、6或甚至3小时，在抑制刺激的降钙素基因相关肽分泌方面非常有效。这些数据提供了第一个证据，肉毒杆菌毒素A型可以直接减少降钙素基因相关肽从三叉神经元释放的量。结果表明，A型肉毒杆菌毒素治疗偏头痛的有效性可能部分是由于其抑制降钙素基因相关肽从激活的感觉神经元释放的能力。
Objective.-To determine the effect of botulinum toxin type A on calcitonin gene-related peptide secretion from cultured trigeminal ganglia neurons.Background.-The ability of botulinum toxins to cause muscle paralysis by blocking acetylcholine release at the neuromuscular junction is well known. Previous studies and clinical observations have failed to demonstrate sensory changes related to botulinum toxins or the disease of botulism. Recent studies, however, have suggested that botulinum toxin type A injected into pericranial muscles may have a prophylactic benefit in migraine. This observation has renewed the debate of a mechanism of sensory inhibition mediated by botulinum toxin type A.Methods.-Primary cultures of rat trigeminal ganglia were utilized to determine whether botulinum toxin type A could directly decrease the release of calcitonin gene-related peptide, a neuropeptide involved in the underlying pathophysiology of migraine. Untreated cultures or cultures stimulated with a depolarizing stimulus (potassium chloride) or capsaicin, an agent known to activate sensory C fibers, were treated for 3, 6, or 24 hours with clinically effective doses of botulinum toxin type A or a control vehicle. The amount of calcitonin gene-related peptide secreted into the culture media following the various treatments was determined using a specific radioimmunoassay.Results.-A high percentage (greater than 90%) of the trigeminal ganglia neurons present in 1- to 3-day-old cultures was shown to express calcitonin gene-related peptide. Treatment with depolarizing stimuli (potassium chloride), a mixture of inflammatory agents, or capsaicin caused a marked increase (4- to 5-fold) in calcitonin gene-related peptide released from the trigeminal neurons. Interestingly, overnight treatment of trigeminal ganglia cultures with therapeutic concentrations of botulinum toxin type A (1.6 or 3.1 units) did not affect the amount of calcitonin gene-related peptide released from these neurons. The stimulated release of calcitonin gene-related peptide following chemical depolarization with potassium chloride or activation with capsaicin, however, was greatly repressed by the botulinum toxin, but not by the control vehicle. A similar inhibitory effect of overnight treatment with botulinum toxin type A was observed with 1.6 and 3.1 units. These concentrations of botulinum toxin type A are well within or below the range of tissue concentration easily achieved with a local injection. Incubation of the cultures with toxin for 24, 6, or even 3 hours was very effective at repressing stimulated calcitonin gene-related peptide secretion when compared to control values.Conclusions.-These data provide the first evidence that botulinum toxin type A can directly decrease the amount of calcitonin gene-related peptide released from trigeminal neurons. The results suggest that the effectiveness of botulinum toxin type A in the treatment of migraine may be due, in part, to its ability to repress calcitonin gene-related peptide release from activated sensory neurons.