经皮三叉神经慢性电刺激对慢性癫痫大鼠海马和皮层内VGLUT1与VGAT表达的影响

发布时间：2018-05-09 14:21

本文选题：癫痫 + 三叉神经电刺激　；参考：《安徽医科大学》2014年硕士论文

【摘要】：目的观察经皮三叉神经慢性电刺激(trigeminal nerve chronic stimulation，TCNS)对匹罗卡品诱导的慢性癫痫大鼠海马和皮层内囊泡型谷氨酸转运体-（1vesicularglutamate transporter1, VGLUT1）与囊泡型γ-氨基丁酸转运体（vesicular GABAtransporter，VGAT）表达的影响，探讨三叉神经电刺激治疗癫痫的可能机制。方法选用成年健康雄性SD大鼠150只，随机选出100只建立匹罗卡品慢性癫痫模型，然后将达到癫痫持续状态（Status epilepticus，SE）的实验大鼠随机分为模型(Pilo)组和经皮三叉神经电刺激(TNS)组；正常对照（Control）组给予同剂量的生理盐水代替匹罗卡品腹腔注射，随后Control组和Pilo组均给予为期一个月的经皮三叉神经假性电刺激、TNS组给予为期一个月的真性电刺激。各组大鼠均于经皮三叉神经电刺激结束后24h、72h、7d、14d、28d取材，通过免疫荧光双标和Western blot方法，，分析海马和皮层内VGLUT1和VGAT的表达情况。所得数据以均数±标准差(x±s)表示，组间均数比较采用单因素方差(One-way ANOVA)分析,以P0.05为差异有统计学意义，P0.01为差异有显著统计学意义。结果 1.VGLUT1在海马和皮层内的表达 TNS组和Pilo组大鼠海马、皮层内VGLUT1的表达较Control组均明显增加，总体上均呈先升高后下降的趋势，约在72h左右达到高峰。在24h时，TNS组大鼠海马和皮层内VGLUT1的表达均明显高于Pilo组（P0.01）,而在以后各个时间点均低于Pilo组（P0.01，P0.05）；TNS组大鼠海马内VGLUT1的表达量在28d时、皮层内在14d和28d时与Control组相比差异无统计学意义（P0.05）,但与Pilo组相比差异均有统计学意义（P0.05）。 2.VGAT在海马和皮层内的表达 TNS组和Pilo组大鼠海马、皮层内VGAT的表达较Control组均明显增加，在72h之前VGAT的表达量逐渐增加，约在72h左右达到高峰，随后开始下降。在72h时，TNS组大鼠海马和皮层内VGAT的表达量均明显低于Pilo组（P0.01）,而在24h、7d、14d、28d时均明显高于Pilo组（P0.01，P0.05）。在28d时，Pilo组大鼠海马和皮层内VGAT的表达量与Control组相比差异无统计学意义（P0.05）、但与TNS组相比差异仍有统计学意义（P0.05）。结论在匹罗卡品诱导的慢性癫痫大鼠模型形成过程中，予以经皮三叉神经慢性电刺激处理减弱了脑的兴奋性机制、增强了其内源性抑制性机制，从而使脑的兴奋易感性降低；这一作用机制可能与脑内囊泡型谷氨酸转运体-1先增多后减少、囊泡型γ-氨基丁酸转运体明显增多有关，这可能是三叉神经电刺激减少癫痫发作的机制之一。
[Abstract]:Purpose The effects of trigeminal nerve chronic stimulation on the expression of vesicular-glutamate transporter-1 (VGLUT1) and vesicular GABA transporter-1 (VGLUT1) in hippocampus and cortex of pilocarpine induced chronic epilepsy rats were observed. To explore the possible mechanism of trigeminal nerve stimulation in the treatment of epilepsy. Method A total of 150 adult male Sprague-Dawley (SD) rats were randomly selected to establish pilocarpine chronic epilepsy model, and then the experimental rats with status epilepticus were randomly divided into two groups: model Pilo group and percutaneous trigeminal nerve stimulation group (TNSgroup). The normal control group was given the same dose of normal saline instead of pilocarpine intraperitoneal injection, then the Control group and the Pilo group were given one month of percutaneous trigeminal nerve pseudoelectric stimulation for one month. The expression of VGLUT1 and VGAT in hippocampus and cortex were analyzed by immunofluorescence double labeling and Western blot method at 24 h, 72 h, 7 d, 14 d and 28 d after the end of percutaneous trigeminal nerve stimulation. The data were expressed as mean 卤standard deviation (x 卤s). One-way ANOVAanalysis was used to compare the mean between groups. There was a significant difference between the two groups (P0.05) (P 0.01). Result Expression of 1.VGLUT1 in hippocampus and cortex The expression of VGLUT1 in hippocampus and cortex of TNS group and Pilo group was significantly higher than that of Control group, and the expression of VGLUT1 increased first and then decreased, reaching the peak at about 72 h. At 24 hours, the expression of VGLUT1 in hippocampus and cortex was significantly higher in the group of Pilo than that in the group of Pilo, and the expression of VGLUT1 in the hippocampus was lower than that in the group of Pilo at every time point, and the expression of VGLUT1 in the hippocampus of the rats in the group of Pilo was lower than that in the group of P0.01and P0.05 at 28d. There was no significant difference between the cortex and the Control group on the 14th and 28th days, but there was significant difference between the cortex and the Pilo group (P 0.05). Expression of 2.VGAT in hippocampus and cortex The expression of VGAT in hippocampus and cortex of TNS group and Pilo group was significantly higher than that of Control group. The expression of VGAT increased gradually before 72 h, reached the peak at about 72 h, then began to decrease. At 72 h, the expression of VGAT in hippocampus and cortex of rats in TNS group was significantly lower than that in Pilo group (P 0.01), and was significantly higher than that in Pilo group at 14 d (14 d) at 24 h. On the 28th day, the expression of VGAT in hippocampus and cortex of rats in the Pilo group was not significantly different from that in the Control group (P 0.05), but there was still a significant difference compared with the TNS group. Conclusion In the course of the formation of pilocarpine induced chronic epilepsy rat model, the chronic electrical stimulation of the trigeminal nerve weakened the excitability mechanism of brain, enhanced its endogenous inhibitory mechanism, and reduced the susceptibility to brain excitability. This mechanism may be related to the increase of vesicular glutamate transporter -1 and the decrease of vesicular type 纬 -aminobutyric acid transporter, which may be one of the mechanisms of trigeminal electrical stimulation to reduce epileptic seizures.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R742.1

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