甲醛刺激断乳期大鼠造成中枢系统FOS蛋白的变化
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摘要
目的:观察断乳期大鼠、成年大鼠右后足背受甲醛刺激后形成兴奋性,由脊髓上传至海马、前皮层;并受NR2A亚单位阻断剂NVP-AAM077和NR2B亚单位阻断剂ifenprodil影响的情况。方法:检测机械性刺激缩足阈值(PWT),检查受甲醛刺激前后脊髓背角,海马CA1区和大脑前皮层HE染色阳性区域和免疫组化的FOS蛋白阳性区域和Western Blot条带。结果:相同伤害性刺激条件下,断乳期和成年大鼠PWT、HE染色、FOS蛋白的免疫组化和Western Blot显示三个部位同步兴奋性增高,在给予ifenprodil后又抑制了兴奋,而成年大鼠ifenprodil和NVP-AAM077皆抑制兴奋的作用。结论:NMDA受体NR2B亚单位抑制剂ifenprodil,可显著减轻甲醛刺激引起断乳期大鼠的炎性疼痛。
Objective: To study the FOS protein changes in central nvervous system stimulated by formaldehyde in weaning rats.Methods: The right hind paws of the weaning rats(P20-P38) were stimulated by formaldehyde to form excitability, which was uploaded from the spinal cord to the hippocampus and anterior cortex; and this excitability was affected by the NR2 A subunit blocker NVP-AAM077 and NR2 B subunit blocker ifenprodil. The mechanical stimulation threshold(PWT) was measured; the number of HE staining positive cells in the spinal dorsal horn, hippocampal CA1 region and anterior cortex was examine; FOS protein expressions were detected by immunustaining and Western Blot before and after stimulation with formaldehyde.Results: Under the same noxious stimulation conditions, PWT, HE staining, immunohistochemistry of FOS protein and Western Blot showed increased excitability at three sites of central nvervous system in rats in weaning period and adulthood,while inhibited excitability was observed after administration of ifenprodil, but NR2 A only had an inhibitory effect on adult rats.Conclusion: Inflammatory pain can be caused by formaldehyde stimulation in weaning rats, and ifenprodil inhibiting NMDA receptor NR2 B subunit can significantly reduce inflammatory pain and impove central nervous system changes.
Objective: To study the FOS protein changes in central nvervous system stimulated by formaldehyde in weaning rats.Methods: The right hind paws of the weaning rats(P20-P38) were stimulated by formaldehyde to form excitability, which was uploaded from the spinal cord to the hippocampus and anterior cortex; and this excitability was affected by the NR2 A subunit blocker NVP-AAM077 and NR2 B subunit blocker ifenprodil. The mechanical stimulation threshold(PWT) was measured; the number of HE staining positive cells in the spinal dorsal horn, hippocampal CA1 region and anterior cortex was examine; FOS protein expressions were detected by immunustaining and Western Blot before and after stimulation with formaldehyde.Results: Under the same noxious stimulation conditions, PWT, HE staining, immunohistochemistry of FOS protein and Western Blot showed increased excitability at three sites of central nvervous system in rats in weaning period and adulthood,while inhibited excitability was observed after administration of ifenprodil, but NR2 A only had an inhibitory effect on adult rats.Conclusion: Inflammatory pain can be caused by formaldehyde stimulation in weaning rats, and ifenprodil inhibiting NMDA receptor NR2 B subunit can significantly reduce inflammatory pain and impove central nervous system changes.
引文
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[2] Wang LZ, Wei CN, Xiao F, et al. Incidence and risk factors for chronic pain after elective caesarean delivery under spinal anaesthesia in a Chinese cohort:a prospective study[J]. Int J Obstet Anesth, 2018,34:21-27.
[3] Ferland CE, Vega E, Ingelmo PM. Acute pain management in children:challenges and recent improvements[J]. Curr Opin Anaesthesiol, 2018, 31(3):327-332.
[4] Price TJ, Inyang KE. Commonalities between pain and memory mechanisms and their meaning for understanding chronic pain[J]. Prog Mol Biol Transl Sci, 2015,131:409-434.
[5] Liu Y, Zhou LJ, Wang J, et al. TNF-αdifferentially regulates synaptic plasticity in the hippocampus and spinal cord by microglia-dependent mechanisms after peripheral nerve injury[J]. J Neurosci, 2017, 37(4):871-881.
[6]户雪雪,彭慧浈,程小娥,等.米诺环素缓解大鼠炎性痛的脊髓机制[J].中华医学杂志,2017, 97(32):2 538-2 543.Hu XX, Peng HZ, Cheng XE, et al. Spinal analgesic mechanism of minocycline in formalin-induced inflammatory pain[J]. National Medical Journal of China,2017,97(32):2 538-2 543.
[7] Sch?fer S, Berger JV, Deumens R, et al. Influence of intrathecal delivery of bone marrow-derived mesenchymal stem cells on spinal inflammation and pain hypersensitivity in a rat model of peripheral nerve injury[J]. J Neuroinflammation, 2014,11:157.
[8] Mangione AS, Obara I, MaiarúM,et al. Nonparalytic botulinum molecules for the control of pain[J]. Pain,2016,157(5):1 045-1 055.
[9] Wlaschin JJ, Gluski JM, Nguyen E,et al. Dual leucine zipper kinase is required for mechanical allodynia and microgliosis after nerve injury[J]. Elife, 2018,7:1-19.
[10] Tochiki KK, MaiarúM, Norris C, et al. The mitogen and stress-activated protein kinase 1 regulates the rapid epigenetic tagging of dorsal horn neurons and nocifensive behaviour[J]. Pain, 2016,157(11):2 594-2 604.
[11] Thomson LM, Zeng J, Terman GW. An N-methyl-Daspartate receptor mediated large, low-frequency, spontaneous excitatory postsynaptic current. in neonatal rat spinal dorsal horn neurons[J] Neuroscience, 2006, 141(3):1 489-1 501.
[12] Zhang XM, Yan XY, Zhang B, et al. Activity-induced synaptic delivery of the GluN2A-containing NMDA receptor is dependent on endoplasmic reticulum chaperone Bip and involved in fear memory[J]. Cell Res, 2015,25(7):818-836.
[13] Guo W, Wei F, Zou S, et al. Group I metabotropic glutamate receptor NMDA receptor coupling and signaling cascade mediate spinal dorsal horn NMDA receptor2B tyrosine phosphorylation associated with inflammatory hyperalgesia[J]. J Neurosci, 2004, 24(41):9 161-9 173.
[14] Han M, Kim J. Effect of dietary iron loading on recognition memory in growing rats[J]. PLoS One, 2015,10(3):1-20.
[15] Greenhill SD, Juczewski K, de Haan AM, et al. NEURODEVELOPMENT. Adult cortical plasticity depends on an early postnatal critical period[J]. Science,2015,349(6246):424-427.