眶下神经干周围注射滑石粉建立新型大鼠三叉神经痛动物模型的实验研究

发布时间：2018-02-08 12:28

本文关键词： 动物模型三叉神经痛滑石粉大鼠　出处：《青岛大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:建立一种适合临床研究的、简单、易行、可靠的三叉神经痛动物模型。方法:实验一选取Wistar雄性大鼠20只,随机分为两组,一组在眶下神经孔周围注射30㳠滑石粉混悬液0.3ml,另一组注射同等剂量的生理盐水,于术前3天以及术后3天、1周、2周、3周、4周、6周、8周、12周进行机械痛阈行为学测定。观察大鼠眶下神经支配区域对机械刺激的疼痛反应阈值及疼痛行为学表现。实验二选取Wistar雄性大鼠20只,随机分为两组,一组在眶下神经孔周围注射30㳠滑石粉混悬液0.3ml,另一组注射同等剂量的生理盐水,于术前3天以及术后3天、1周、2周、4周进行视频行为学测定。摄像机放置在笼子前面1米处观察大鼠探究行为、静止行为、抓脸行为、摇头行为、舔后肢行为和舔身体行为的次数和时间。实验三选取Wistar大鼠20只,分为滑石粉注射组和对照组,每组各10只。按照实验1的方法制作滑石粉三叉神经痛模型。分别于术后3天、4周、8周、12周取大鼠眶下神经、延髓组织作组织病理学观察及脱髓鞘染色,应用免疫组化的方法检测眶下区组织肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)以及延髓中疼痛物质(p物质、内啡肽等)的表达。结果:实验一实验组大鼠于术后3天出现眶下神经支配区域机械痛反应阈值与术前及对照组相比明显降低(P㩳0.01),大鼠易激惹,具有搔抓面部或攻击行为。机械痛阈值降低持续至术后12周。实验二视频显示:实验组大鼠于术后3天出现探究行为、静止行为、抓脸行为、摇头行为的变化,与术前及对照组相比上述行为变化有统计学意义(P㩳0.01),舔后肢、舔身体行为无明显变化。大鼠术后1周、2周、4周的探究行为、静止行为、抓脸行为、摇头行为变化进一步明显,与术前及对照组相比上述行为变化均有统计学意义(P㩳0.01)。舔后肢、舔身体行为与术前及对照组相比上述行为变化无统计学意义。实验三实验组术后3天组织病理学观察:主要是炎症表现,炎性因子表达少.术后1周炎症更剧烈免疫组化出现炎性因子表达.术后4周炎性因子表达最高,4-12周炎性反应逐渐减轻,局部瘢痕加重,可见压迫眶下神经,脱髓鞘染色可见脱髓鞘表现;延髓组织中疼痛物质的表达于3天-3周无明显变化,术后4-12周出现疼痛物质表达增加并出现脱髓鞘表现。结论:1、眶下神经干注射滑石粉可致眶下神经支配区域出现痛觉超敏现象,自发性疼痛以及相关行为学改变;2、组织病理学及免疫组化检查显示该动物模型早期为炎性因子引起疼痛,晚期为眶下神经压迫致病。3、该动物模型可早期诱导疼痛且持续时间长,简单易行,为目前较理想的三叉神经痛动物模型。
[Abstract]:Objective: to establish a simple, easy and reliable animal model of trigeminal neuralgia suitable for clinical study. Methods: in experiment 1, 20 male Wistar rats were randomly divided into two groups. The talc powder suspension was 0.3 ml, and the other group was injected with the same dose of normal saline. The mechanical pain threshold was measured 3 days before operation and 3 weeks after surgery. The threshold of pain response to mechanical stimulation in the innervation region of the suborbital nerve and the behavior of pain behavior in the suborbital nerve innervation region of rats were observed. The mechanical pain threshold was measured at the end of 4 weeks, 6 weeks and 8 weeks and 12 weeks after operation. Experiment two selected 20 male Wistar rats, They were randomly divided into two groups. One group was injected around the suborbital foramen with 30? The talc powder suspension was 0.3 ml. Another group was injected with the same dose of normal saline. The video behavior was measured 3 days before operation and 3 days after operation. The camera was placed in front of the cage to observe the exploring behavior and static behavior of the rats. The number and time of face scratching, shaking head, licking hind limbs and licking body behavior. Experiment 3 selected 20 Wistar rats and divided them into talcum powder injection group and control group. The model of trigeminal neuralgia with talc powder was made according to the method of experiment 1 in each group. The suborbital nerve was taken from rats at 3 days, 4 weeks, 8 weeks and 12 weeks after operation, and the medulla oblongata was observed by histopathological observation and demyelinating staining. Tumor necrosis factor- 伪 (TNF- 伪), interleukin-1 尾 (IL-1 尾) and pain substance in medulla oblongata were detected by immunohistochemical method. Results: the mechanical pain response threshold of the suborbital innervation region in experimental group 1 was significantly lower than that in preoperation and control group. The mechanical pain threshold was decreased until 12 weeks after operation. Experiment 2 video showed that the rats in the experimental group showed the changes of inquiry behavior, static behavior, face grabbing behavior and shaking head behavior 3 days after operation. Compared with the preoperative and control groups, the changes of the above behaviors were statistically significant (P < 0.05). There was no significant change in the behavior of lapping hind limbs and licking the body. The behavior of inquiry, static behavior, face grabbing and shaking head were significantly changed in rats 1 week and 2 weeks after operation, compared with those before operation and control group. There was no significant difference in the behavior between the experimental group and the control group. The histopathological changes were observed 3 days after operation in the experimental group: inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation, inflammation. The expression of inflammatory factors was less. 1 week after operation, inflammatory factors were expressed more intensely by immunohistochemistry. The highest expression of inflammatory factors was observed at 4 to 12 weeks after operation. The inflammatory reaction gradually decreased and the local scar became more severe, and the suborbital nerve was compressed. Demyelination staining showed demyelination, and the expression of pain substance in medulla oblongata did not change significantly at 3 weeks after 3 days. At 4-12 weeks after operation, the expression of pain substance increased and demyelination appeared. Conclusion injection of talcum powder into the suborbital nerve trunk can induce hyperalgesia in the innervation region of the suborbital nerve. Spontaneous pain and related behavioral changes. Histopathological and immunohistochemical examination showed that the animal model was caused by inflammatory factors at the early stage and by suborbital nerve compression at the late stage. The animal model could induce pain early and last for a long time. It is an ideal animal model of trigeminal neuralgia.
【学位授予单位】：青岛大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R745.11;R-332

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