【摘要】：目的：通過正中神經電刺激（Median nerve electrical stimulation，MNS）TBI后昏迷動物模型，研究MNS對大鼠前額葉皮質（Prefrontal cortex，PFC）Orexin-A及OX1R表達的影響，初步探討MNS對TBI后昏迷促醒的作用及可能機制。 方法：將大鼠分為空白組、假刺激組、刺激組，建立持續(xù)性腦外傷后昏迷大鼠模型并評估意識狀態(tài)，給予MNS后，再次評估意識狀態(tài)。各組分別于電刺激后6h、12h、24h每個時間點均處死20只，快速分離PFC組織，采用酶聯(lián)免疫法（Enzyme-linked immunoassay，ELISA）、蛋白質印跡(Western Blot)、免疫組化（Immunohistochemistry，IHC）技術檢測大鼠PFC區(qū)Orexin-A及OX1R的表達水平變化。 結果： ELISA檢測表明刺激組PFC區(qū)Orexin-A水平升高，與空白組、假刺激組相比差異有統(tǒng)計學意義（P＜0.05），且6h、12h、24h刺激組PFC區(qū)Orexin-A水平呈上升的趨勢，各個時間點刺激組與空白組、假刺激組相比差異有統(tǒng)計學意義（P＜0.05）；Western Blot檢測結果表明三組PFC區(qū)OX1R的水平差異有統(tǒng)計學意義（P＜0.05），其中刺激組PFC區(qū)OX1R水平升高，與空白組、假刺激組相比差異統(tǒng)計學意義（P＜0.05），三組PFC區(qū)OX1R水平在6h、12h、24h呈上升趨勢，6h、12h刺激組與假刺激組相比，差異有統(tǒng)計學意義（P＜0.05），24h刺激組與假刺激組相比，差異無統(tǒng)計學意義（P＞0.05）；IHC檢測結果表明檢測表明刺激組PFC區(qū)OX1R水平與空白組和假刺激組相比差異有統(tǒng)計學意義（P＜0.05）。6h，12h，，24h三組PFC區(qū)OX1R水平呈上升趨勢，且差異有統(tǒng)計學意義（P＜0.05）。6h三組PFC區(qū)OX1R水平差異無統(tǒng)計學意義（P＞0.05），而12h和24h差異有統(tǒng)計學意義（P＜0.05）。 結論：1、MNS對TBI后昏迷大鼠有促醒作用，其機制可能通過上調大鼠PFC區(qū)Orexin-A和OX1R水平； 2、TBI后昏迷大鼠PFC區(qū)Orexin-A和OX1R水平在損傷后出現(xiàn)緩慢上升的趨勢，在24h達到一個高值，考慮其可能是正常生理性的應急反應，以保護受損的神經元，降低神經元的損害。
[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.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2014
【分類號】：R741

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1 黃強;戴偉民;揭園慶;余國峰;范曉峰;吳安;呂堯;李運平;;重型顱腦損傷昏迷患者持續(xù)右正中神經刺激促進復蘇的臨床研究[J];中國臨床神經科學;2011年03期

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