Changes of CGRP8-37-induced Antinociception and CGRP-immunoreactivity at Spinal Levels in Morphine Tolerant Rats

神经药理学报 ›› 2013, Vol. 3 ›› Issue (2): 1-12.

• 研究论文 • 下一篇

Changes of CGRP8-37-induced Antinociception and CGRP-immunoreactivity at Spinal Levels in Morphine Tolerant Rats

YANG Ying, YU Long-Chuan

北京大学生命科学学院神经生物学实验室，生物膜与膜生物工程国家重点实验室，北京 100871，中国

出版日期:2013-04-26 发布日期:2014-06-27
通讯作者: 于龙川，男，北京大学生命科学学院教授，博士生导师；研究方向：痛觉的调节机制，药物耐受与成瘾的机制；Tel: 86-10-62753667, E-mail: yulc@pku.edu.cn
作者简介:杨莹，女，美国杜克大学博士生
基金资助:
国家自然科学基金（30470542，30870802，8117043）,君政基金（20040001057）,国家重点基础研究发展计划（973）项目（2009CB522002）

Changes of CGRP8-37-induced Antinociception and CGRP-immunoreactivity at Spinal Levels in Morphine Tolerant Rats

YANG Ying, YU Long-Chuan

Neurobiology Laboratory and National Laboratory of Biomembrane and Membrane Biotechnology, College of Life Sciences, Peking University, Beijing 100871, China

Online:2013-04-26 Published:2014-06-27
Contact: 于龙川，男，北京大学生命科学学院教授，博士生导师；研究方向：痛觉的调节机制，药物耐受与成瘾的机制；Tel: 86-10-62753667, E-mail: yulc@pku.edu.cn
About author:杨莹，女，美国杜克大学博士生
Supported by:
国家自然科学基金（30470542，30870802，8117043）,君政基金（20040001057）,国家重点基础研究发展计划（973）项目（2009CB522002）

摘要/Abstract

摘要： 目的：研究吗啡耐受后大鼠脊髓水平降钙素基因相关肽抑制剂CGRP8-37镇痛效应的变化，以及对降钙素基因相关肽表达的影响。方法：在大鼠脊髓蛛网膜下腔内埋管，应用热板法和Randall Selitto Test测量大鼠对伤害性热刺激和机械刺激引起的后爪缩爪反应潜伏期(hindpaw withdrawl lantency, HWL)作为痛觉反应指标, 应用免疫组织化学的方法观察吗啡耐受前、吗啡耐受后及耐受恢复后大鼠脊髓背角和背根神经节中降钙素基因相关肽免疫活性物质的变化。结果： (1) 证实在脊髓蛛网膜下腔注射10 nmol的CGRP8-37后，大鼠对伤害性热刺激和机械刺激的缩爪反应潜伏期显著增加，表明在正常大鼠脊髓水平CGRP8-37具明确的镇痛作用。（2）在吗啡耐受大鼠蛛网膜下腔注射10 nmol CGRP8-37后，大鼠对伤害性热刺激和机械刺激的缩爪反应潜伏期显著增加，表明吗啡耐受大鼠脊髓水平CGRP8-37具有明确的镇痛作用。在吗啡耐受恢复后，脊髓水平CGRP8-37具有镇痛作用。(3) 与正常大鼠相比，在吗啡耐受大鼠蛛网膜下腔注射同样剂量的CGRP8-37所产生的镇痛作用有显著性下降，而在耐受恢复后，其作用基本恢复到正常水平。(4) 应用免疫组化方法，我们证实在L3-L5段脊髓背角第I、II层中存在大量CGRP免疫活性纤维，在脊髓L3-L5段背根神经节中存在大量的小到中等大小的CGRP免疫阳性的神经元。(5) 在吗啡耐受后，L3-L5段脊髓背角和L3-L5段背根神经节中的CGRP免疫活性物质的含量明显升高；耐受恢复后，背根神经节中CGRP免疫活性物质的含量基本恢复到正常水平，脊髓背角浅层中CGRP免疫活性物质的含量虽然有所下降，但仍稍高于正常水平。结论: 在吗啡耐受后，大鼠脊髓水平CGRP8-37的镇痛作用及CGRP的含量都发生了可塑性变化，提示CGRP在吗啡耐受中具有重要作用。

Abstract: Objective: It is known that intrathecal administration of calcitonin gene-related peptide 8-37 (CGRP8-37), an antagonist to CGRP1 receptor, induced antinociception. The present study was performed to investigate the changes in nociceptive modulation of CGRP8-37 and CGRP-immunoreactivity at spinal levels after morphine tolerance. Methods: The hindpaw withdrawal latencies (HWLs) to both thermal and mechanical stimulation were assessed by hot plate and Randall Selitto Test. The CGRP-like immunoreactivity at spinal levels was tested by immunohistochemistry. Results: The hindpaw withdrawal latencies (HWLs) to both thermal and mechanical stimulation increased significantly after intrathecal injection of 10 nmol of CGRP8-37. There were also significant increases in HWLs to both thermal and mechanical stimulation after intrathecal administration of CGRP8-37 in morphine tolerant rats and rats recovered from morphine tolerance. It is interesting that the CGRP8-37-induced antinociceptive effects were lower in morphine tolerant rats than those in morphine naive rats and rats recovered from morphine tolerance. Furthermore, there were increases in CGRP-like immunoreactivity in dorsal horn of the spinal cord and the dorsal root ganglia (DRG) of L3 – L5 in morphine tolerant rats. Conclusion: The results of the present study demonstrated the down-regulations in CGRP8-37-induced antinociception after morphine tolerance, which is possibly resulted from the changes in both the opioid and the CGRP systems. The latter was implicated partly in the present study, in that there were plastic changes in CGRP-like immunoreactivity in dorsal horn and DRG after morphine tolerance. All results above suggested that CGRP might play an important role in morphine tolerance at the spinal levels.

Key words: antinociception, calcitonin gene-related peptide (CGRP), CGRP8-37, morphine tolerance, spinal cord, dorsal root ganglia (DRG)

YANG Ying, YU Long-Chuan. Changes of CGRP8-37-induced Antinociception and CGRP-immunoreactivity at Spinal Levels in Morphine Tolerant Rats [J]. 神经药理学报, 2013, 3(2): 1-12.

YANG Ying, YU Long-Chuan. Changes of CGRP8-37-induced Antinociception and CGRP-immunoreactivity at Spinal Levels in Morphine Tolerant Rats [J]. Acta Neuropharmacologica, 2013, 3(2): 1-12.

参考文献

[1] Yu Long-chuan, Hou Jun-feng, Fu Feng-hua, et al. Roles of calcitonin gene-related peptide and its receptors in pain-related behavioral responses in the central nervous system[J]. Neurosci Biobehv Rev, 2009, 33(8): 1185–1191.

[2] Russell F A, King R, Smillie S J, et al. Calcitonin gene-related Peptide: physiology and pathophysiology[J]. Physiol Rev, 2014, 94(4): 1099-1142.

[3] Denise van Rossum, Uwe Karsten Hanisch, Remi Quirion. Neuroanatomical localization, pharmacological characterization and functions of CGRP, related peptides and their receptors[J]. Neurosci Biobehav Rev, 1997, 21(5): 649-678.

[4] 刘芳薇, 洪炎国. 降钙素基因相关肽家族在疼痛和阿片耐受中的作用[J].生理学报, 2013, 65(3):347-354.

[5] Stephen G Howitt, Kalle Kilk, Wang Yang, et al. The role of the 8-18 helix of CGRP8-37 in mediating high affinity binding to CGRP receptors, coulombic and steric interactions[J]. Br J Pharmacol, 2003, 138(2): 325-332.

[6] Francois B Jolicoeur, Daniel Menard, Alain Fournier, et al. Structure-activity analysis of CGRP’s neurobehavioral effects[J]. Ann NY Acad Sci, 1992, 657:155-163.

[7] Yu Long-chuan, Per Hansson, Tomas Lundeberg. The calcitonin gene-related peptide antagonist CGRP8-37 increases the latency to withdrawal responses in rats[J]. Brain Res, 1994, 653(1-2):223-230.

[8] Yu Long-chuan, Per Hansson, Gunilla Brodda-Jansen, et al. Intrathecal CGRP8-37-induced bilateral increase in hindpaw withdrawal latency in rats with unilateral inflammation [J]. Br J Pharmacol, 1996, 117(1):43-50.

[9] Yu Long-chun, Per Hansson, Lundeberg T. The calcitonin gene-related peptide antagonist CGRP8-37 increases the latency to withdrawal responses bilaterally in rats with unilateral experimental mononeuropathy, an effect reversed by naloxone[J]. Neuroscience, 1996, 71(2):523–531.

[10] Yu Yi, Thomas Lundeberg, Yu Long-chuan. Role of calcitonin gene-related peptide and its antagonist on the evoked discharge frequency of wide dynamic range neurons in the dorsal horn of the spinal cord in rats[J]. Regul Pept, 2002, 103(1):23-27.

[11] Yu Long-chuan, Per Hansson, Thomas Lundeberg. Opioid antagonists naloxone, beta-funaltrexamine and naltrindole, but not nor-binaltorphimine, reverse the increased hindpaw withdrawal latency in rats induced by intrathecal administration of the calcitonin gene-related peptide antagonist CGRP8-37[J]. Brain Res, 1995, 698(1-2):23-29.

[12] Ballet S, Mauborgne A, Benoliel J J, et al. Polyarthritis-associated changes in the opioid control of spinal CGRP release in the rat[J]. Brain Res, 1998, 796(1):198-208.

[13] Bao Lan, Jin Shan-xue, Zhang Chen, et al. Activation of delta opioid receptors induces receptor insertion and neuropeptide secretion[J]. Neuron, 2003, 37(1):121-133.

[14] Sunita Chakrabarti, Martin Oppermann, Alan R Gintzler. Chronic morphine induces the concomitant phosphorylation and altered association of multiple signaling proteins: a novel mechanism for modulating cell signaling[J]. Proc Natl Acad Sci USA, 2001, 98(7):4209-4214.

[15] Subrata Basu Ray, Himanshu Gupta, Yogendre Kumar Gupta. Up-regulation of m-opioid receptors in the spinal cord of morphine-tolerant rats[J]. J Biosci, 2004, 29(1):51-56.

[16] Zhou Xiang, Li Jin-ju, Yu Long-chuan. Plastic changes of calcitonin gene-related peptide in morphine tolerance: behavioral and immunohistochemical study in rats[J]. J Neurosci Res, 2003, 74(4):622-629.

[17] Kelly J Powell, Weiya Ma, Maaja Sutak, et al. Blockade and reversal of spinal morphine tolerance by peptide and non-peptide calcitonin gene-related peptide receptor antagonists[J]. Br J Pharmacol, 2000, 131(5):875-884.

[18] Zimmermann M. Ethical guidelines for investigations of experimental pain in conscious animals[J]. Pain, 1983, 16(2):109-110.

[19] Li J J, Zhou X, Yu L C. Involvement of neuropeptide Y and Y1 receptors in antinociception in the arcuate nucleus of hypothalamus, an immunohistochemical and pharmacological study in intact rats and rats with inflammation[J]. Pain, 2005, 118(1-2):232-242.

[20] Sun Yan-gang, Gu Xing-long, Yu Long-chuan. The neural pathway of galanin in the hypothalamic arcuate nucleus of rats: Activation of beta-endorphinergic neurons projecting to periaqueductal gray matter[J]. J Neurosci Res, 2007, 85(11):2400-2406.

[21] Tony L Yaksh, Thomas A Rudy. Chronic catheterization of the spinal subarachnoid space[J]. Physiol Behav, 1976, 17(6):1031-1036.

[22] Mizushima T, Obata K, Yamanaka H, et al. Activation of p38 MAPK in primary afferent neurons by noxious stimulation and its involvement in the development of thermal hyperalgesia[J]. Pain, 2005, 113(1-2):51-60.

[23] Multon S, Pardutz A, Mosen J, et al. Lack of estrogen increases pain in the trigeminal formalin model: a behavioural and immunocytochemical study of transgenic ArKO mice[J]. Pain, 2005, 114(1-2):257-265.

[24] Luis R Gardell, Wang Rui-zhong, Shannon E Burgess, et al. Sustained morphine exposure induces a spinal dynorphin-dependent enhancement of excitatory transmitter release from primary afferent fibers[J]. J Neurosci, 2002, 22(15):6747-6755.

[25] Belanger S, Ma W, Chabot J G, et al. Expression of calcitonin gene-related peptide, substance P and protein kinase C in cultured dorsal root ganglion neurons following chronic exposure to mu, delta and kappa opiates[J]. Neuroscience, 2002, 115(2):441-453.

[26] Forrest L Smith, Ruby R Javed, Mark J Elzey, et al. The expression of a high level of morphine antinociceptive tolerance in mice involves both PKC and PKA[J]. Brain Res, 2003, 985(1):78-88.

[27] Wang Zhi-yong, Jean-Guy Chabot, Remi Quirion. On the possible role of ERK, p38 and CaMKII in the regulation of CGRPexpression in morphine-tolerant rats[J]. Mol Pain, 2011, 7:68-.

[28] Wang Z, Ma W, Jean-Guy Chabot, et al. Calcitonin gene-related peptide as a regulator of neuronal CaMKII-CREB, microglial p38-NFκB and astroglial ERK-Stat1/3 cascades mediating the development of tolerance to morphine-induced analgesia[J]. Pain, 2010, 151(1):194-205.

Changes of CGRP8-37-induced Antinociception and CGRP-immunoreactivity at Spinal Levels in Morphine Tolerant Rats

Changes of CGRP8-37-induced Antinociception and CGRP-immunoreactivity at Spinal Levels in Morphine Tolerant Rats

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 1

编辑推荐

Metrics