大鼠胫神经瘤移置模型的建立及其神经系统中神经营养素的表达变化

发布时间：2018-01-24 18:26

本文关键词： 胫神经瘤移置模型神经瘤压痛神经营养素神经生长因子脑源性神经营养因子　出处：《上海交通大学学报(医学版)》2017年01期 　论文类型：期刊论文

【摘要】：目的·建立大鼠胫神经瘤移置(TNT)模型,观察损伤神经水平及背根神经节(DRG)和脊髓内神经营养素(NTs)家族中神经生长因子(NGF)和脑源性神经营养因子(BDNF)的表达变化,初步探讨外周神经损伤后NTs表达变化对神经瘤形成和疼痛相关行为学的影响。方法·将大鼠胫神经横断并经皮下隧道移置在外踝上方皮下,建立TNT模型。于术后不同时间点应用Von Frey纤维丝测量神经瘤压痛和足底机械刺激反应阈值(PWT);实验终点处死大鼠,取损伤胫神经断端,Neurofi lament-200荧光标记神经纤维观察胫神经断端神经瘤形成情况。再次造模,并分别于第7、21、42、49日处死大鼠,取损伤侧胫神经、DRG和脊髓,ELISA检测胫神经和DRG中NGF和BDNF的表达变化,Western blotting检测脊髓中NGF和BDNF的表达变化。结果·术后第7日,神经瘤压痛反应频率和评分显著升高;术后第3日,足底外侧缘PWT开始降低且神经瘤压痛和足底外侧机械痛持续整个实验周期;但足底中部PWT无明显变化。胫神经免疫荧光染色表明损伤神经断端神经瘤形成。ELISA检测结果显示:胫神经内NGF表达增加,BDNF在术后第7、21日表达升高后逐渐降至正常水平;DRG内NGF在术后第7、21日表达升高后逐渐降至正常水平,BDNF在术后第7日表达升高后逐渐降至正常水平。在脊髓水平,在术后不同时间点NGF和BDNF的表达均升高并持续整个实验周期。结论·TNT可导致大鼠损伤神经断端神经瘤形成及神经病理性疼痛(神经瘤压痛和足底机械痛觉高敏),其发生和发展过程可能与NGF和BDNF有关。
[Abstract]:Objective: to establish TNT model of rat tibial neuroma. The level of injured nerve and the expression of nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF) in DRG (dorsal root ganglion) and neurotrophin (NTs) family in spinal cord were observed. Objective: to investigate the effect of NTs expression on neuroma formation and pain related behavior after peripheral nerve injury. Methods: the tibial nerve was transected and subcutaneous tunnel was placed above the lateral malleolus. The TNT model was established. Von Frey fibril was used to measure the tenderness of neuroma and the threshold of plantar mechanical stimulation at different time points after operation. The rats were killed at the end of the experiment. Neurofi lament-200 labeled nerve fibers were taken from the injured tibial nerve to observe the formation of neuroma at the broken end of the tibial nerve. The rats were killed on the 7th day at the 21st day and the rats were killed on the 9th day. The expression of NGF and BDNF in the tibial nerve and DRG were detected by Elisa and the tibial nerve was taken from the injured side of the tibial nerve. Western blotting was used to detect the expression of NGF and BDNF in spinal cord. Results: on the 7th after operation, the frequency and score of tenderness response were significantly increased. On 3rd, PWT began to decrease in the lateral margin of the plantar and the tenderness of neuroma and mechanical pain on the lateral plantar continued throughout the experimental cycle. But there was no obvious change of PWT in the middle of plantar. The expression of NGF in the tibial nerve was increased by immunofluorescence staining of tibial nerve. The expression of BDNF decreased gradually to normal level at the 7th and 21st postoperatively. The expression of NGF in DRG gradually decreased to normal level on the 7th postoperative and 21st, and then decreased to normal level on 7th after operation, and then decreased to normal level at spinal cord level. The expression of NGF and BDNF increased at different time points after operation and lasted for the whole experiment cycle. Conclusion: TNT can induce neuroma formation and neuropathic pain in injured rats. Neuroma tenderness and plantar mechanical pain Gao Min. Its occurrence and development may be related to NGF and BDNF.
【作者单位】：上海交通大学附属第六人民医院疼痛科;
【基金】：国家自然科学基金(81370933,81400803,81672237)~~
【分类号】：R739.4;R-332
【正文快照】： 疼痛是截肢后的常见并发症,临床表现为幻肢痛和残肢痛。在经受截肢手术的打击之后,约80%的截肢患者出现幻肢痛;另外,约有61%的患者遭受残肢痛的困扰[1-2]。目前认为,截肢后疼痛的发病机制可能与断端神经瘤形成有关[3-5]。损伤的外周神经有强大的再生能力。在其再生过程中,神

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