CGRP相关新型偏头痛药物研究进展

<正>1概述偏头痛是一种慢性致残性疾病,表现为持续4～72 h的中至重度头痛且反复发作,影响着全球高达15%的人口~([1])。在20世纪90年代,偏头痛治疗的重大突破是曲坦类药物的发现。曲坦类药物是特异性5-羟色胺(5-Ht 1b和5-Ht 1 d)受体选择性激动剂,被认为是现有最好的急性偏头痛的特异性治疗药物,它们通过使血管收缩来发挥治疗偏头痛的作     

Chemically distinct rat olivocochlear neurons.

We have produced a neurochemical map of the cell bodies of origin of the cochlear efferent terminals in rat by combining glutamic acid decarboxylase (GAD), choline acetyltransferase (ChAT), or calcitonin gene-related peptide (CGRP) immunocytochemistry with retrograde transport of horseradish peroxidase. The locations of cochlear efferent cell bodies are in general agreement with the medial and lateral systems described by White and Warr (J. Comp. Neurol. 219:203-214, 1983) with some minor modifications. The lateral system consists of at least two pools of chemically distinct neurons located within the lateral superior olive (LSO) ipsilateral to the injected cochlea. One pool immunostains with an antibody to GAD while the other immunostains with antibodies to ChAT and to CGRP. The medial efferent system consists of periolivary neurons that are almost exclusively large and ChAT-positive but CGRP-negative. They are located both ipsilateral and contralateral to the cochlea they project to. There are a few GAD-positive small neurons in the medioventral and rostral periolivary regions that project ipsilaterally, but these may prove tobe ectopic neurons. The ipsilateral lateroventral periolivary region (LVPO) contains some efferent neurons, all of which are ChAT-positive but CGRP-negative. Additional cochlear efferent neurons, some of which are ChAT-positive and others GAD-positive, are present within and immediately dorsal to the fiber capsule surrounding the medial limb, and to a lesser extent the lateral limb, of the ipsilateral LSO. Not all GAD-positive or ChAT-positive olivary cells project to the cochlea. We have complemented the results in the brainstem by demonstrating two immunocytochemically distinct populations of efferent terminals in the cochlea simultaneously, one CGRP-positive and the other GAD-positive. Approximately equal numbers of boutons immunoreactive for both markers are present beneath inner hair cells throughout the entire length of the cochlea. Surprisingly high numbers of GAD-positive and CGRP-positive boutons are also present on outer hair cells, with each class having its spatially and morphologically distinct features. The lack of CGRP-positive periolivary cells that are retrogradely labeled by cochlear injections of HRP suggests that the lateral olivocochlear system sends projections to outer hair cells. Our results raise questions about species differences in the organization of targets of the lateral and medial olivocochlear systems.     

The isolated spinal cord-skin preparation of the newborn rat and effects of some algogenic and analgesic substances

We have developed an isolated spinal cord-skin preparation of the newborn rat. The spinal cord together with a piece of skin connected to the cord by the saphenous nerve was isolated from 1- to 4-day-old rats and separately superfused with artificial cerebrospinal fluid in two neighbouring chambers. Potentials were recorded extracellularly from the third lumbar ventral root. Application of capsaicin (0.5-2 μM) or KCl (60–350 mM) with brief pressure pulses to the perfusion bath of the skin evoked a depolarizing response of 20- to 40-s duration in the ventral root. The response was depressed by [Met⁵]enkcphalin (0.03–3 μM). morphine (0.1–2 μM) and a tachykinin antagonist, [D-Arg¹,D-Trp^7,9,Leu¹¹] substance P (spantide), 1–10 μM), applied to the spinal cord by superfusion, whereas the response was augmented by centrally administered calcitonin gene-related peptide (0.1–0.2 μM) or bicuculline (0.5–1 μM).     

Calcitonin gene-related peptide-like immunoreactivity, in botulinum toxin-paralysed rat muscles

Changes in calcitonin gene-related peptide-like immunoreactivity (CGRP-LI) at the motor endplates of botulinum toxin-paralysed rat muscles were investigated using immunohistochemistry. One day following toxin injection, a dramatic increase in CGRP-LI was detected at the motor endplates and within preterminal axons of the soleus and gastrocnemius muscles. The upregulation of CGRP-LI persisted throughout the period during which muscle fibres were paralysed and new neuromuscular junctions were being formed by the growing sprouts. Decline of CGRP-LI at the motor endplates coincided with clinical recovery. Both up- and down-regulation of CGRP-LI took place earlier in the soleus than in the gastrocnemius muscle. Up-regulation of CGRP-LI was also detected in a subpopulation of motor axons in the sciatic nerves and in the spinal motor neurons innervating the paralysed muscles. These results indicate that levels of CGRP are regulated, at least partly, by changes in the target innervation. They also suggest an important role for CGRP in the regenerative processes following muscle paralysis.     

Ascending general visceral pathways within the brainstems of two teleost fishes: Ictalurus punctatus and Carassius auratus.

The primary general visceral nucleus in goldfish (Carassius auratus) and catfish (Ictalurus punctatus) is located at the ventroposterior boundary of the vagal gustatory lobe and receives coelomic visceral, but not gustatory inputs. The neuronal tracer horseradish peroxidase (HRP) was employed to visualize sources of input to and ascending projections from the primary general visceral nucleus in these species. In addition, immunocytochemical techniques were utilized to define the cytological divisions within the pontine gustatory-visceral complex. The pontine secondary visceral nuclei in both catfish and goldfish contains numerous somata and fibers immunoreactive for calcitonin gene-related peptide (CGRP). In contrast, the secondary gustatory nuclei are devoid of fibers and cells immunoreactive for CGRP. In both the goldfish and the channel catfish, the primary general visceral nucleus receives input from the vagal gustatory lobe, as well as the medullary reticular formation. In the channel catfish, the primary general visceral nucleus projects bilaterally to the secondary visceral nucleus, which lies rostrolateral to the secondary gustatory nucleus in the dorsal pons. Fibers cross the midline via the rostral part of the isthmic commissure. Injection of HRP into the primary general visceral nucleus of a goldfish labels ascending fibers that project to a secondary visceral nucleus situated ventral, lateral, and rostral to the secondary gustatory complex. In general, the results indicate that general visceral systems ascend in parallel to gustatory systems within the brainstem, and that general visceral but not gustatory nuclei are immunoreactive for the peptide CGRP.     

血浆ET-1和CGRP含量测定对脑出血病人的应用价值

目的 :探讨血浆内皮素 (ET)和降钙素基因相关肽 (CGRP)在脑出血病人急性期与恢复期中的变化。方法 :采用放射免疫分析技术 ,对 4 2例脑出血病人的急性期与恢复期 ,分别测定血浆ET和CGRP含量 ,并用 35例健康对照人员作比较。结果 :脑出血病人血浆ET含量明显高于对照组 (p <0 0 1) ,治疗后有所下降 ;而血浆CGRP含量 ,在脑出血的急性期 ,显著高于对照组 (p <0 0 1) ,恢复期降为正常。结论 :血浆ET和CGRP在脑出血的急性期显著升高 ,并与脑损伤程度有关 ,提示神经肽参与出血性脑损伤的发生发展过程     

Caudal end of the rat spinal dorsal horn

We have previously demonstrated that the transformation of the caudal spinal cord through the conus medullaris to the filum terminale takes place in three steps. In the conus medullaris the twin layers of CGRP-immunoreactive and IB4-labeled primary afferent fibers as well as the translucent portion of the superficial dorsal horn equivalent to the substantia gelatinosa discontinue before the complete removal of the dorsal horn. Parallel with these changes VGLUT1-immunoreactive myelinated primary afferent fibers arborize not only in the deep layers but also in the entire extension of the remaining dorsal horn, while scattered CGRP fibers still remains at the margin of and deep in the dorsal horn. PKCgamma-immunoreactive dorsal horn neurons discontinue parallel with the disappearance of the IB4-labeled nerve fibers. These observations suggest that in the dorsal horn certain neurons are linked to the substantia gelatinosa, while others are substantia gelatinosa-independent neurons.     

Substance P and calcitonin gene-related peptide: immunohistochemical localisation and microvascular effects in rabbit skeletal muscle

Summary 1. The distribution and microvascular effects of substance P (SP) and calcitonin gene-related peptide (CGRP) were studied in the rabbit tenuissimus muscle using immunohistochemistry and intravital microscopy. 2. Individual fibers within nerve bundles and along blood vessels in the muscle were found to be immunoreactive (IR) for both SP and CGRP, thus showing an apparently complete coexistence for these peptides. In dorsal root ganglia most SP-positive cells were also CGRP-IR, but the latter cells were somewhat more numerous than SP-IR cells. 3. When applied topically to the muscle, both SP and CGRP increased blood flow in a dose-dependent manner, but CGRP was more potent and caused responses of longer duration. Both SP and CGRP dilated transverse arterioles, but they had little or no effect on the smaller terminal arterioles. This resulted in a redistribution of blood flow to the connective tissue adjacent to the muscle. 4. SP, but not CGRP, elicited vigorous vasomotion in larger arterioles and caused the formation of aggregates of platelets and leukocytes in the venules. Neither flow increase, nor vasomotion or aggregate formation were influenced by pretreatment of the animals with mepyramine, cimetidine or indomethacin. Capsaicin (1 M) had a powerful effect on transverse arterioles resembling that of both SP and CGRP. 5. It is concluded that some of the vascular effects hitherto ascribed to SP on the basis of nerve stimulation and application of capsaicin might, at least in part, be due to release of CGRP. Send offprint requests to: A. Ohlen, Department of Physiology, Karolinska Institutet, S-104 01 Stockholm, Sweden     

慢性肾衰患者外源性PGE1与血浆内皮素及降钙素基因相关肽的关系

目的观察外源性前列腺素E1(PGE1)对慢性肾衰(CRF)患者血浆内皮素 (ET)及降钙素基因相关肽(CGRP)水平的影响. 方法利用放射免疫分析法测定9 0例CRF患者经PGE1治疗后的血浆ET及CGRP水平的变化并分析其相互关系. 结果 CRF患者血浆ET的水平较对照组明显升高(P<0.01) ,血浆CGRP的水平较对照组明显降低(P<0.01). ET与肾功能参数 (BUN,Cr)呈明显的正相关(r=0.58 ,P<0.01;r=0.62,P<0.01),与尿量呈明显的负相关(r=-0.60,P< 0.01). PGE1治疗后,ET和CGRP水平均有所好转(P<0.05),同时肾功能参数(BUN, Cr) 和尿量亦有改善(P<0.05). 结论 CRF多伴有血浆ET及CGRP 水平的变化,应用PGE1治疗有益于延缓肾功能的进一步恶化.     

Nitroglycerin-inhibited whole blood aggregation is partially mediated by calcitonin gene-related peptide–a neurogenic mechanism

1. The role of the vasculature and calcitonin gene-related peptide (CGRP) in nitroglycerin (NTG)-mediated platelet inhibition was studied.
2. In vitro incubations of CGRP in whole blood induced a dose-dependent inhibition of platelet aggregation with an IC₅₀ of 62.1 nM.
3. The platelet inhibition induced by CGRP was blocked by co-incubation of 0.53 μM CGRP_8-37, as well as 30 μM N^G-nitro-monomethyl-L-arginine (L-NMMA).
4. In a separate group of experiments, 100 nM NTG in rat whole blood (WB) induced platelet inhibition of 6.0±1.3% (mean±s.d.), which was enhanced to 77.6±3.5% by the addition of rat aortic tissue (AT) (P<0.001). The inclusion of CGRP_8-37 with NTG and AT in WB reduced platelet inhibition to 31.6±6.8% (P<0.01). Incubation of WB and AT with 30 μM L-NMMA reduced NTG-induced inhibition of platelet aggregation to 26.4±4.2% (P<0.001).
5. It is concluded that vascular tissue contributes to the antiplatelet mechanism of action of NTG. Furthermore, NTG apparently evokes the release of CGRP from vascular tissue and this neuropeptide contributes to the antiplatelet actions of NTG.
6. The antiplatelet activity of CGRP in whole blood is mediated primarily through the activation of nitric oxide synthase.