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論文名稱 Title |
活化血清素神經系統對於慢性頸部脊髓損傷後呼吸功能恢復之影響 Effects of serotonergic activation on the respiratory functional recovery following chronic cervical spinal cord injury |
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系所名稱 Department |
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畢業學年期 Year, semester |
語文別 Language |
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學位類別 Degree |
頁數 Number of pages |
74 |
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研究生 Author |
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指導教授 Advisor |
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召集委員 Convenor |
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口試委員 Advisory Committee |
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口試日期 Date of Exam |
2015-07-16 |
繳交日期 Date of Submission |
2015-08-25 |
關鍵字 Keywords |
血清素2A亞型接受器、頸部第2節脊髓半邊切除、血清素、呼吸、橫膈肌 5-HT2A receptor, C2Hx, diaphragm, serotonin, ventilation |
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統計 Statistics |
本論文已被瀏覽 5706 次,被下載 220 次 The thesis/dissertation has been browsed 5706 times, has been downloaded 220 times. |
中文摘要 |
頸部脊髓半邊切除會阻斷延腦向下投射的呼吸傳遞路徑,導致損傷側橫膈肌癱瘓,呼吸功能喪失。研究指出血清素神經系統參與急性頸部脊髓損傷後橫膈神經的活性誘發,但對於慢性頸部損傷後呼吸功能的影響不明瞭,本論文欲探討慢性頸部脊髓損傷後活化血清素神經系統對於潮氣容積與橫膈肌活性的調控作用。將雄性Sprague-Dawley大鼠進行頸部第2節脊髓半邊切除手術,實驗流程分為組織學與生理學測量兩部分:(1) 組織學:損傷後8週將動物進行免疫螢光與免疫化學染色,免疫螢光染色前塗抹逆向追蹤劑標定橫膈運動神經元,觀察頸部第3-4節脊髓血清素分布;(2) 生理學:損傷後8週測量大鼠自發性呼吸下呼吸氣流及橫膈肌電圖 (electromyogram,EMG),並於靜脈注射血清素前驅物5-羥色胺酸 (5-hydroxy-tryptophan,5-HTP,10 mg/kg) 、血清素回收抑制劑 (fluoxetine,10 mg/kg) 或血清素2A亞型接受器致效劑 (TCB-2,0.05 mg/kg) 活化血清素神經系統。組織學顯示脊髓損傷後損傷側第3-4節頸部脊髓血清素含量降低。生理學發現損傷組的潮氣容積與損傷側橫膈肌放電強度低於未損傷組,在處理5-HTP或TCB-2後,損傷側橫膈肌強度顯著上升,但潮氣容積無明顯變化,然而處理fluoxetine對損傷側橫膈肌與潮氣容積皆無影響。由上述結果可推論頸部脊髓損傷後8週脊髓橫膈運動神經元周圍血清素含量仍然下降,藉由增加血清素含量或活化血清素2A亞型接受器可誘發損傷側橫膈肌放電活性,然而仍不足以改善整體呼吸功能。 |
Abstract |
Cervical spinal cord hemisection interrupts the bulbospinal respiratory pathway and leads to ipsilateral hemidiaphragm paralysis and respiratory dysfunction. Previous studies have reported that serotonergic activation can induce partial recovery of phrenic activity after acute cervical hemisection at 2nd cervical spinal cord (C2Hx). Nevertheless, it is unknown whether activation of the serotonergic system can still modulate respiratory function during the chronic state. The aim of the present study was to investigate whether serotonergic system activation facilitates the respiratory functional recovery following chronic cervical spinal cord injury. Adult male Sprague-Dawley rats received C2Hx or C2 laminectomy were used for histological and physiological study after 8 weeks post-injury: (1) The phrenic motoneurons were labeled by a retrograde tracer (Cholera toxin subunit B) and expression of serotonin (5-HT) fibers at 3-4th cervical spinal cord were detected by immunohistochemical staining. (2) Tidal volume and diaphragm muscle electromyogram (EMG) activity were recorded in anesthetized and spontaneously breathing rats. The results demonstrated 5-HT was reduced in the ipsilateral to the lesion following C2Hx. The tidal volume and ipsilateral diaphragm EMG activity were decreased at 8 weeks post-injury compared with uninjured. Both 5-HTP and TCB-2 induced a significant and persistent increased in the ipsilateral diaphragm amplitude. However, the tidal volume was not changed. In addition, administration of fluoxetine did not influence diaphragm EMG and tidal volume. These results suggest that exogenous supplement with serotonin and activation of 5-HT2A receptor can increase the ipsilateral diaphragm activity; however, these approaches are not sufficient to improve respiratory functional recovery following chronic cervical spinal cord injury. |
目次 Table of Contents |
論文審定書+i 中文摘要+ii Abstract+iii 圖次+vi 表次+vii 縮寫表+viii 第一章 前言+1 1.1 頸部脊髓損傷對於呼吸功能的影響+1 1.2 血清素神經系統+3 1.3 目的+9 第二章 材料與方法+11 2.1 實驗動物與分組+11 2.2 頸部脊髓損傷手術+11 2.3 血清素藥物配置+12 2.4 生理參數測量+12 2.5 逆向標記橫膈運動神經元+13 2.6 脊髓組織處理與冷凍切片+14 2.7 損傷組織染色與殘餘面積定量+14 2.8 免疫化學染色+14 2.9 免疫螢光染色+15 2.10 數據分析與統計+16 第三章 結果+18 3.1 頸部第2節脊髓損傷程度+18 3.2 慢性頸部脊髓損傷對於血清素表現之影響+18 3.3 慢性頸部脊髓損傷對於呼吸型態、心跳、血壓與橫膈肌放電活性之影響+18 3.4 5-HTP對於慢性頸部脊髓損傷後呼吸型態、心跳、血壓與橫膈肌放電活性之影響+19 3.5 Fluoxetine對於慢性頸部脊髓損傷後呼吸型態、心跳、血壓與橫膈肌放電活性之影響+20 3.6 TCB-2對於慢性頸部脊髓損傷後呼吸型態、心跳、血壓與橫膈肌放電活性之影響+21 第四章 討論+23 4.1 方法討論+23 4.2 慢性頸部脊髓損傷後對於呼吸功能的影響+24 4.3 慢性頸部脊髓損傷後活化血清素神經系統對於呼吸功能的調節+24 4.4 結論+30 第五章 圖與說明+32 第六章 表與說明+46 第七章 參考文獻 +48 第八章 附錄+57 8.1 補充數據+57 8.2 會議論文摘要與壁報+62 |
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