正中神经电刺激对脑外伤后昏迷大鼠前额叶皮质Orexin-A和OX1R表达的影响

发布时间：2018-09-05 16:05

【摘要】：目的：通过正中神经电刺激（Median nerve electrical stimulation，MNS）TBI后昏迷动物模型，研究MNS对大鼠前额叶皮质（Prefrontal cortex，PFC）Orexin-A及OX1R表达的影响，初步探讨MNS对TBI后昏迷促醒的作用及可能机制。方法：将大鼠分为空白组、假刺激组、刺激组，建立持续性脑外伤后昏迷大鼠模型并评估意识状态，给予MNS后，再次评估意识状态。各组分别于电刺激后6h、12h、24h每个时间点均处死20只，快速分离PFC组织，采用酶联免疫法（Enzyme-linked immunoassay，ELISA）、蛋白质印迹(Western Blot)、免疫组化（Immunohistochemistry，IHC）技术检测大鼠PFC区Orexin-A及OX1R的表达水平变化。结果： ELISA检测表明刺激组PFC区Orexin-A水平升高，与空白组、假刺激组相比差异有统计学意义（P＜0.05），且6h、12h、24h刺激组PFC区Orexin-A水平呈上升的趋势，各个时间点刺激组与空白组、假刺激组相比差异有统计学意义（P＜0.05）；Western Blot检测结果表明三组PFC区OX1R的水平差异有统计学意义（P＜0.05），其中刺激组PFC区OX1R水平升高，与空白组、假刺激组相比差异统计学意义（P＜0.05），三组PFC区OX1R水平在6h、12h、24h呈上升趋势，6h、12h刺激组与假刺激组相比，差异有统计学意义（P＜0.05），24h刺激组与假刺激组相比，差异无统计学意义（P＞0.05）；IHC检测结果表明检测表明刺激组PFC区OX1R水平与空白组和假刺激组相比差异有统计学意义（P＜0.05）。6h，12h，，24h三组PFC区OX1R水平呈上升趋势，且差异有统计学意义（P＜0.05）。6h三组PFC区OX1R水平差异无统计学意义（P＞0.05），而12h和24h差异有统计学意义（P＜0.05）。结论：1、MNS对TBI后昏迷大鼠有促醒作用，其机制可能通过上调大鼠PFC区Orexin-A和OX1R水平； 2、TBI后昏迷大鼠PFC区Orexin-A和OX1R水平在损伤后出现缓慢上升的趋势，在24h达到一个高值，考虑其可能是正常生理性的应急反应，以保护受损的神经元，降低神经元的损害。
[Abstract]:Aim: to study the effect of MNS on the expression of Orexin-A and OX1R in the prefrontal cortex (Prefrontal cortex,PFC) of rats after TBI (median nerve stimulation) TBI, and to explore the effect and possible mechanism of MNS on the promotion of coma after TBI. Methods: the rats were divided into blank group and sham stimulation group. The model of coma rats after persistent brain injury was established and the conscious state was evaluated. After MNS, the conscious state was evaluated again. 20 rats were killed at each time point at 12 h and 24 h after electrical stimulation. PFC tissues were separated quickly. The expression of Orexin-A and OX1R in PFC region of rats were detected by enzyme linked immunosorbent assay (Enzyme-linked immunoassay,ELISA) and Western blot (Western Blot), immunohistochemistry (Immunohistochemistry,IHC). Results: ELISA showed that the level of Orexin-A in the PFC region of the stimulation group was higher than that of the blank group and the sham stimulation group (P < 0. 05), and the Orexin-A level in the PFC region of the stimulation group at 6 h or 12 h and 24 h after stimulation showed an increasing trend, and at each time point, the level of Orexin-A in the PFC region of the stimulation group was higher than that in the blank group. The results of Western Blot analysis showed that the level of OX1R in the PFC region of the three groups was significantly higher than that in the control group (P < 0. 05), and the level of OX1R in the PFC region of the stimulation group was higher than that in the blank group (P < 0. 05). The level of OX1R in the PFC region of the three groups showed an increasing trend at 6 h and 12 h after stimulation (P < 0. 05). There was no significant difference between the two groups (P > 0. 05). The level of OX1R in the three groups was not significantly higher than that in the sham stimulation group (P < 0. 05). (2) there was no significant difference between the two groups (P > 0. 05), and there was no significant difference between the two groups (P < 0. 05). The results of IHC test showed that the level of OX1R in the PFC region of the stimulated group was significantly higher than that of the blank group and the sham stimulation group (P < 0. 05). The OX1R level in the PFC region of the three groups showed an upward trend in 24 hours after 12 hours of stimulation (P < 0. 05). There was no significant difference in OX1R level among the three groups (P < 0. 05), but there was a significant difference between 12 h and 24 h (P < 0. 05). ConclusionTwo one MNS can promote the wake-up of the coma rats after TBI, the mechanism may be by upregulating the levels of Orexin-A and OX1R in the PFC region of the rats, and the Orexin-A and OX1R levels in the PFC area of the rats after TBI may increase slowly after injury, and reach a high value at 24 hours. Consider that it may be a normal physiological emergency response to protect damaged neurons and reduce neuronal damage.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R741

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