發(fā)布時間：2018-04-29 13:00

  本文選題：脊髓損傷 + 膈肌功能��； 參考：《第二軍醫(yī)大學》2015年碩士論文

【摘要】：[目的]脊髓膈神經(jīng)元胞體所在核團主要位于頸3-頸5水平,高位頸脊髓損傷病人,若損傷累及頸3-頸5水平,隨之則出現(xiàn)完全性或不完全性的膈肌癱瘓。受損的呼吸系統(tǒng)相關(guān)肌肉嚴重限制了患者的活動、生活質(zhì)量及發(fā)聲,因而在慢性進展階段不得不采用機械通氣以輔助患者呼吸。然而伴隨機械通氣所帶來的并發(fā)癥逐漸增加,如呼吸機相關(guān)性肺炎、張力性氣胸及縱膈氣腫等等。長期機械通氣引起的呼吸道相關(guān)并發(fā)癥是導致高位脊髓損傷患者高死亡率高主要原因。本課題組發(fā)現(xiàn)可以將大鼠副神經(jīng)作為動力神經(jīng),移位吻合至膈神經(jīng)以重建高位頸髓橫斷傷后膈肌運動功能。本課題擬在其基礎(chǔ)上建立單側(cè)頸3脊髓半切動物模型,進一步探討神經(jīng)移位術(shù)后大鼠膈肌功能恢復的程度,并通過神經(jīng)逆行示蹤的方法,研究呼吸相關(guān)神經(jīng)通路重塑的的情況。[方法]健康成年雄性S-D大鼠(100-150克)48只,平均隨機分配到A、B、C三組,每組16只。A組為健康大鼠組,B組為左側(cè)膈神經(jīng)、副神經(jīng)在鎖骨水平離斷組,C組為左側(cè)副神經(jīng)斜方肌支鎖骨水平移位吻合膈神經(jīng)干組。飼養(yǎng)6個月后,每組隨機選取8只大鼠行胸片、肺功能、動脈血氣分析,之后將這部分大鼠處死。剩余每組8只大鼠行頸3脊髓左側(cè)半切損傷,飼養(yǎng)1周后,再次將每組剩余8只大鼠,行胸片、肺功能、動脈血氣分析。再次選取健康成年雄性S-D大鼠(100-150克)24只,隨機平均分配至D、E、F三組,每組8只。其中D、E組為健康大鼠組,F組為神經(jīng)移位組,正常飼養(yǎng)6月后,E、F組大鼠行頸3脊髓左側(cè)半切。左側(cè)膈肌行綠色熒光蛋白基因重組的偽狂犬病毒定點注射,順時針選取4點,每點注射5微升。正常飼養(yǎng)4天后行心臟灌注、取腦。連續(xù)冰凍切片后于熒光顯微鏡觀察,并同大鼠腦解剖圖譜進行比對。選取帶有熒光的切片行免疫組織化學染色。使用NIS-Elements2.1軟件獲取圖像,使用Image-Pro Plus 6.0行綠色熒光顯色陽性神經(jīng)元胞體計數(shù)。統(tǒng)計學分析采用SPSS 20.0軟件,采用單因素方差分析進行組間比較。組間兩兩比較采用Bonferroni法(方差齊時)或Mann-Whitney法(方差不齊時),P=0.05。[結(jié)果]頸3左側(cè)脊髓半切前,B、C兩組大鼠肺功能相關(guān)指標均低于A組,但C組優(yōu)于B組。頸3左側(cè)脊髓半切后,A組大鼠各項肺功能指標約下降50%,B、C兩組指標同脊髓半切前基本持平,略有下降,此時C組大鼠各項指標均優(yōu)于A、B兩組。E組大鼠未觀察到熒光顯像的神經(jīng)元細胞,D、F組大鼠在延髓內(nèi)檢測到綠色熒光,免疫組織化學染色證實為綠色熒光蛋白基因重組偽狂犬病毒轉(zhuǎn)染的神經(jīng)元細胞核團。[結(jié)論]副神經(jīng)斜方肌支在鎖骨水平移位吻合膈神經(jīng)主干的方法可以部分重建頸3至頸5水平頸脊髓損傷大鼠的膈肌功能,同時可以一定程度上改善大鼠的呼吸功能。通過逆行示蹤的方式發(fā)現(xiàn)神經(jīng)移位術(shù)重新建立了膈肌同延髓之間的聯(lián)系,此神經(jīng)通路在頸脊髓損傷大鼠膈肌功能恢復中起到了重要的作用。
[Abstract]:[objective] the nucleus of the phrenic neuron is located mainly at the level of cervical 3 neck 5. If the injury involves the level of 3 neck 5 of the cervical cord, complete or incomplete diaphragmatic paralysis will occur in the patients with high cervical spinal cord injury. Damaged respiratory muscles severely limit the patient's activity, quality of life, and voice, so mechanical ventilation has to be used to assist the patient in the chronic stage of progression. However, complications associated with mechanical ventilation, such as ventilator-associated pneumonia, tension pneumothorax and mediastinal emphysema, are increasing. Respiratory tract related complications caused by long-term mechanical ventilation are the main cause of high mortality in patients with high spinal cord injury. Our team found that the accessory nerve can be used as the dynamic nerve and anastomosed to the phrenic nerve to reconstruct the diaphragm motor function after high cervical spinal cord transection. Based on the model of unilateral cervical 3 spinal cord hemisectomy, this study aims to explore the degree of recovery of diaphragm function in rats after nerve transposition, and to study the remodeling of respiratory related nerve pathway by retrograde tracing of nerve. [methods] Forty-eight healthy adult male S-D rats were randomly assigned to group A (n = 16), group B (n = 16) and group B (n = 16), with left phrenic nerve in group B (n = 8). The accessory nerve was transfered horizontally from the clavicular branch of the left trapezius muscle to the phrenic trunk group. After 6 months of feeding, 8 rats in each group were randomly selected for chest radiography, pulmonary function and arterial blood gas analysis. The remaining 8 rats in each group were treated with left side hemisection injury of the cervical 3 spinal cord. After feeding for 1 week, the remaining 8 rats in each group were again given chest radiographs, pulmonary function and arterial blood gas analysis. Twenty-four healthy adult male S-D rats were randomly divided into three groups (8 rats in each group). Among them, DU E group was healthy rat group and F group was nerve transposition group. After 6 months of normal feeding, the left side of the cervical 3 spinal cord was cut in the EOF group. The left diaphragm was injected with pseudorabies virus with green fluorescent protein gene recombination, 4 points clockwise, 5 microliters per point. After normal feeding for 4 days, the heart was perfused and brain was taken. Continuous frozen sections were observed by fluorescence microscope and compared with the anatomical map of rat brain. Sections with fluorescence were selected for immunohistochemical staining. NIS-Elements2.1 software was used to obtain images and Image-Pro Plus 6.0 was used to count the cell bodies of green fluorescent positive neurons. Statistical analysis was carried out with SPSS 20.0 software and single factor analysis of variance (ANOVA). The two groups were compared with each other by Bonferroni method (homogeneity of variance) or Mann-Whitney method (P0. 05 when variance was not equal). [results] the relative indexes of lung function in group C were lower than those in group A, but group C was superior to group B. The indexes of lung function in group A decreased about 50% after hemisection of the left side of the cervical 3 spinal cord, and the indexes in group C were almost the same as those in group C before spinal cord hemisection, and decreased slightly. At this time, the indexes of group C were better than those of group A B. Group E. the rats in group D and F had detected green fluorescence in medulla oblongata. It was confirmed by immunohistochemical staining that it was a neuronal nuclear mass transfected by recombinant pseudorabies virus (PRV) with green fluorescent protein gene (GFP). [conclusion] the method of horizontal transposition of trapezius branch of accessory nerve and anastomosis of phrenic nerve trunk can partly reconstruct the diaphragm function of the rats with cervical spinal cord injury from 3 to 5 levels, and improve the respiratory function of the rats to some extent. It is found by retrograde tracing that the connection between diaphragm and medulla oblongata is re-established by nerve transposition, which plays an important role in the recovery of diaphragmatic function in rats with cervical spinal cord injury.
【學位授予單位】：第二軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2015
【分類號】：R651.2

【參考文獻】

相關(guān)期刊論文 前1條

1 劉才棟;蔣文華;;大鼠副神經(jīng)脊髓核在中樞內(nèi)的定位[J];解剖學報;1986年02期

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本文編號：1820036