ERK在偏头痛动物模型中的激活及与肥大细胞脱颗粒间的关系

发布时间：2018-01-01 13:20

  本文关键词：ERK在偏头痛动物模型中的激活及与肥大细胞脱颗粒间的关系　出处：《山西医科大学》2011年硕士论文　论文类型：学位论文

  更多相关文章： 偏头痛 细胞外信号调节激酶 肥大细胞 硬脑膜 三叉神经核尾部 硝酸甘油 大鼠

【摘要】：目的:通过观察硝酸甘油(GTN)诱导的大鼠偏头痛模型中硬脑膜和三叉神经核尾部磷酸化细胞外信号调节激酶(p-ERK)的表达情况,及p-ERK的表达与肥大细胞脱颗粒间的关系,揭示p-ERK在偏头痛发病过程中的病理生理作用,为偏头痛的治疗提供新的实验依据。 方法:将72只雄性SD大鼠随机分为偏头痛模型组(n=54)和空白对照组(n=18),模型组采用皮下注射GTN(10mg/kg)法建立大鼠偏头痛模型,再将模型组大鼠随机分为三个亚组:Compound48/80组(腹腔注射Compound48/80,2mg/kg);SCG+Compound48/80组(腹腔先注射SCG, 10mg/kg后再注射Compound48/80, 2mg/kg)和实验对照组(腹腔注射同等剂量的生理盐水),空白对照组大鼠皮下注射等量生理盐水。应用免疫组织化学染色法观察模型组各亚组大鼠分别在腹腔注射Compound48/80、SCG+Compound48/80、生理盐水的15min、60min、120min后硬脑膜、三叉神经核尾部磷酸化ERK(p-ERK)的表达情况;空白对照组给予等量生理盐水与GTN组对照,也分别在15min、60min、120min后取硬脑膜、三叉神经核尾部观察p-ERK的表达情况。 结果:1)行为学观察:63只雄性SD大鼠注射GTN,诱发偏头痛模型成功57只(成功率为90.4%),造模5～15min左右大鼠出现双耳发红、频繁挠头、烦躁乱动的现象,约40～60min达高峰,此现象持续1～3h,继之大鼠呈现活动减少、倦卧状态。而Compound48/80组和SCG+Compound48/80组频繁挠头、爬笼次数增多等烦躁现象约18～30min时达高峰。空白对照组仅在15min内略有挠头现象,之后趋于正常,且未出现双耳发红、爬笼次数增多等现象。2)镜下观察:免疫组织化学染色发现空白对照组大鼠各时间点硬脑膜、三叉神经核尾部均有少量p-ERK阳性免疫反应产物;模型组各亚组各时间点硬脑膜、三叉神经核尾部均有较空白对照组对应时间点染色深而多的p-ERK阳性免疫反应产物,以模型组Compound48/80亚组的15min组最为显著,且这些阳性免疫反应产物主要分布于脱颗粒的肥大细胞附近。3)统计结果:ERK在模型组各亚组和空白对照组的各时间点硬脑膜和三叉神经核尾部都发生了磷酸化,但空白对照组的各时间点间无显著差异(P0.05);模型组各亚组各时间点p-ERK阳性免疫反应产物都显著高于空白对照组对应时间点(P0.05);模型组Compound48/80亚组各时间点p-ERK的阳性免疫产物显著高于SCG+ Compound48/80组和实验对照组对应时间点(P0.05);模型组各亚组15min组分别显著高于同组的60min组和120min组(P0.05)。 结论:皮下注射GTN后,大鼠硬脑膜和三叉神经核尾部处p-ERK的表达增加,且与肥大细胞脱颗粒的数量呈正相关,但随时间的延长磷酸化的ERK会失活,提示ERK可能参与了偏头痛早期痛觉信号的传导,肥大细胞脱颗粒可能是促使p-ERK表达增加的原因,支持三叉神经血管炎症学说。
[Abstract]:Objective: to observe the expression of extracellular signal regulated kinase (ERK) in dural and trigeminal nucleus of rats with migraine induced by GTNN. The relationship between the expression of p-ERK and mast cell degranulation reveals the pathophysiological role of p-ERK in the pathogenesis of migraine and provides a new experimental basis for the treatment of migraine. Methods: 72 male Sprague-Dawley rats were randomly divided into migraine model group (n = 54) and blank control group (n = 18). In the model group, the migraine model was established by subcutaneous injection of GTNN 10 mg / kg. Then the model group rats were randomly divided into three subgroups:: component 48 / 80 (intraperitoneal injection of Compound48 / 802 mg / kg); SCG Compound48/80 group (SCG was injected intraperitoneally, 10 mg / kg then Compound48/80). 2 mg / kg) and experimental control group (intraperitoneal injection of the same dose of normal saline). The rats in the blank control group were subcutaneously injected with the same amount of normal saline. The rats of each subgroup of the model group were respectively injected with Compound48/80 in the abdominal cavity by immunohistochemical staining. SCG component 48 / 80, normal saline 15 mins, 60 mins, 120 minutes later, dura mater. The expression of phosphorylated ERK p-ERK in the tail of trigeminal nucleus; The same amount of normal saline was given to the blank control group and the control group, and the expression of p-ERK was observed at the end of trigeminal nucleus, and the dura mater was taken 120 minutes after 15 min or 60 min. Results the behavior of 63 male Sprague-Dawley rats was observed by behavior observation. 57 rats were successfully induced migraine (success rate was 90.4%) by injecting GTNN into male Sprague-Dawley rats. The rats showed redness of ears, frequent scratching of their heads and agitation for about 15 minutes, and reached the peak for about 4060 minutes. This phenomenon lasted for 1 minute for 3 hours, followed by a decrease in the activity of the rats. While Compound48/80 group and SCG Compound48/80 group frequently scratched their heads. The number of cage climbing increased and so on, which reached the peak at 1830 minutes. The blank control group only slightly scratched its head within 15 minutes, then tended to normal, and did not appear auricular redness. Observation under microscope: immunohistochemical staining showed that there were a small amount of p-ERK positive immunoreactive products in the dura mater and tail of trigeminal nucleus of rats in the blank control group at every time point. Compared with the blank control group, there were more p-ERK positive immunoreactive products in the dural and the tail of trigeminal nucleus in each subgroup of model group than in the blank control group. The Compound48/80 subgroup of model group was the most significant in 15min group. These immunoreactive products were mainly distributed near degranulated mast cells. 3). The results showed that the phosphorylation of the dura mater and the tail of trigeminal nucleus occurred at all time points in the model group and the blank control group. However, there was no significant difference between different time points in the blank control group (P 0.05). The p-ERK positive immunoreactive products in each subgroup of the model group were significantly higher than those in the blank control group at each time point (P0.05). The positive immunoreactive products of p-ERK in Compound48/80 subgroup of model group were significantly higher than those in SCG Compound48/80 group and experimental control group at each time point. P0.05; The 15min group of each subgroup in the model group was significantly higher than that in the 60min group and 120min group in the same group. Conclusion: after subcutaneous injection of GTN, the expression of p-ERK in dural and trigeminal nucleus of rats was increased, and it was positively correlated with the number of mast cell degranulation. However, phosphorylated ERK will be inactivated over time, suggesting that ERK may be involved in the early migraine pain signal transduction, mast cell degranulation may be the cause of the increase of p-ERK expression. Supports the theory of trigeminal neurovascular inflammation.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R747.2;R-332

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