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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 呂俊宏(June-Horng Lue) | |
dc.contributor.author | Yi-Ju Tsai | en |
dc.contributor.author | 蔡怡汝 | zh_TW |
dc.date.accessioned | 2021-06-13T16:58:17Z | - |
dc.date.available | 2005-04-21 | |
dc.date.copyright | 2005-04-21 | |
dc.date.issued | 2005 | |
dc.date.submitted | 2005-04-12 | |
dc.identifier.citation | 參考文獻
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/39029 | - |
dc.description.abstract | 中文摘要
本實驗室先前的研究結果顯示,正中神經損傷後在楔狀神經核內所產生的神經胜肽Y,與動物受到神經傷害後所續發的神經病變疼痛 (neuropathic pain) 的症狀似乎互有關聯;然而對於神經胜肽Y在大白鼠楔狀神經核及其背根神經節的研究則相當缺乏。因此本實驗利用正中神經截斷的神經瘤模式,並配合免疫細胞化學方法、神經追蹤劑、背側神經根切斷術、電刺激、電子顯微鏡技術及不同藥劑處理,來探討正中神經截斷損傷後神經胜肽Y在大白鼠楔狀神經核及其背根神經節的來源、突觸組成及減緩神經胜肽Y表現量變化之相關研究。 首先,我們利用螢光金神經追蹤劑標誌正中神經在背根神經節的主要分佈節段,發現正中神經主要分佈在第六及第七頸髓的背根神經節。進一步探討大白鼠正中神經截斷後存活不同時程,在第六及第七頸髓的背根神經節中神經胜肽Y的表現情形,發現正中神經截斷損傷後第三天,即可在手術側背根神經節中觀察到有少許的神經胜肽Y免疫反應神經元表現,爾後神經胜肽Y免疫反應神經元的數目慢慢增加,並且在神經受傷後四週達到高峰,四週後會慢慢減少,二十週時只可觀察到少量神經胜肽Y免疫反應的神經元。進而探討背根神經節內神經胜肽Y與神經損傷的關係,利用螢光金神經追蹤劑注射於截斷的正中神經內,進而標誌背根神經節內軸突受傷的神經元,並配合抗神經胜肽Y免疫細胞化學反應,進行雙重標誌方法。結果顯示在正中神經截斷損傷後一週、二週和四週,高達99%以上的神經胜肽Y免疫反應神經元是屬於受傷神經元,而且在這三組之間並無統計上的差異;另外,發現有高達54~71%以上的受傷神經元具有神經胜肽Y免疫反應,而且四週組的百分比顯著高於一週及二週組。爾後緊接著將螢光金與螢光紅神經追蹤劑分別注射於楔狀神經核和已截斷的正中神經內,分別標誌背根神經節中投射至楔狀神經核的神經元與軸突受傷的神經元,並且配合抗神經胜肽Y免疫細胞化學反應,進行三重標誌方法。結果顯示正中神經截斷後,在背根神經節中有許多被螢光紅標誌的受傷初級感覺神經元具有神經胜肽Y免疫反應而且同時被螢光金所標誌。另外經背側神經根切斷處理後的實驗動物,再進行正中神經截斷損傷,該側楔狀神經核並沒有觀察到任何神經胜肽Y免疫反應產物的表現,顯示楔狀神經核的神經胜肽Y免疫反應神經纖維是源自於初級感覺神經元的初級傳入神經纖維。緊接著,我們使用山葵過氧化氫酵素逆向神經追蹤劑及免疫細胞化學法分別標誌楔狀神經核內投射至丘腦的接轉神經元以及正中神經截斷後所產生的神經胜肽Y,爾後將含有楔狀神經核之組織切片經電子顯微鏡包埋技術處理後,在電子顯微鏡下觀察,發現具有神經胜肽Y免疫反應的神經終末會與楔狀丘腦接轉神經元的樹突形成突觸。經由以上的實驗證明,當正中神經截斷損傷後四週,在第六及第七頸髓的背根神經節中受傷的神經元會產生大量的神經胜肽Y,然而這些反應產物會經由神經元的初級傳入神經纖維投射至楔狀神經核內,並且藉由與楔狀丘腦接轉神經元的樹突所形成之突觸,屆時將訊息傳遞至視丘中。 當正中神經截斷損傷後,受傷側楔狀神經核會產生大量的神經胜肽Y免疫反應神經纖維與c-Fos免疫反應神經元,並且表現區域與時間點都十分相似,為了釐清神經胜肽Y免疫反應神經纖維與c-Fos免疫反應神經元的關係,本實驗使用神經胜肽Y拮抗劑注射於楔狀神經核吻段區域內,並且對已進行截斷四週的正中神經施以電刺激,發現c-Fos免疫反應神經元的數目顯著低於未注射神經胜肽Y拮抗劑之組別。為了進一步瞭解,減少的c-Fos免疫反應神經元是屬於何種類型神經元,本實驗使用螢光金神經追蹤劑注射於丘腦腹側基底核群,用來標誌楔狀神經核內投射至丘腦的接轉神經元,並配合抗c-Fos免疫細胞化學反應,進行雙重標誌方法。結果顯示未注射神經胜肽Y拮抗劑的組別,c-Fos免疫反應神經元為楔狀丘腦接轉神經元的比率 (c-Fos + CTN/c-Fos%) 高達75%,但注射神經胜肽Y拮抗劑後則降至44%,並且具有統計上的差異,由此看來神經胜肽Y拮抗劑會減少楔狀丘腦接轉神經元表現c-Fos。最後,我們在進行正中神經截斷手術前半小時分別使用腎上腺素α2受體競爭劑 (Clonidine)、NMDA受體阻斷劑 (MK-801)、局部麻醉劑 (Lidocaine) 及神經活性阻斷劑 (Tetrodotoxin, TTX) 處理,進而比較手術側楔狀神經核內神經胜肽Y的變化,以期望能提供減緩神經病變疼痛的有效措施。結果顯示先前經局部麻醉劑及神經活性阻斷劑處理的大白鼠,其神經胜肽Y免疫反應神經纖維所佔據楔狀神經核面積之百分比以及神經胜肽Y免疫反應神經纖維光密度值之百分比均有顯著的降低。由此看來,局部麻醉劑及神經活性阻斷劑均能有效抑制神經胜肽Y的產生。 綜合以上所述,周邊神經受到損傷後,神經胜肽Y在背根神經節內只表現在受傷的初級感覺神經元,並透過初級傳入神經纖維投射至楔狀神經核內,使得楔狀丘腦接轉神經元產生過度的神經興奮繼而表現c-Fos,而可能進一步參與神經病變疼痛的訊息傳遞。另外,使用不同藥劑前處理,能有效抑制神經胜肽Y產生,可以提供避免手術所引起神經病變疼痛的有效預防措施。 | zh_TW |
dc.description.abstract | 英文摘要
Our previous study had revealed that the neuropeptide may contribute to nerve injury-induced hyperalgesia in rats’ forelimb of median nerve injury with special reference to neuropeptide Y (NPY). Although several studies has revealed that the increased NPY in the gracile nucleus and dorsal root ganglion (DRG) following sciatic nerve injury, and it still lacks confirmation in the cuneate nucleus (CN) and DRG after complete median nerve transection (CMNT). In the present proposal, we will use the median nerve injury animal models, immunocytochemistry, neuronal tracer labeling, dorsal rhizotomy, electrical stimulation, electromicroscopic, and pre-emptive drug treatment in anesthetized rats. First, we examined characteristics of the neuropeptide Y-like immunoreactive (NPY-LI) DRG neurons after CMNT. With Fluorogold (FG) injection into normal median nerves, numerous FG-labeled DRG neurons were predominantly localized in the C6 and C7 DRGs, where focal regions were examined after CMNT. Along with NPY immunohistochemistry, a few NPY-LI neurons were detected in the ipsilateral but not contralateral DRGs after FG injection into the nerve. As early as three days after CMNT a few NPY-LI neurons could be detected, reaching a maximum in the percentage of the NPY-LI neurons in the DRGs at four weeks, subsided thereafter over twenty weeks. The NPY-LI DRG neurons were primarily medium-sized and large neurons. With FG injection into the transected median nerve, we found that about 99% of NPY-LI neurons were labeled for FG, suggesting that they are derived from the injured but not intact DRG neurons. Then, using double fluorescent dyes tract tracing, we detected that some of the injured DRG neurons were NPY-LI neurons that projected to the CN. Further treated with dorsal rhizotomy, our data indicated that after CMNT the induced NPY-LI fibers in the ipsilateral CN were exclusively originated from the injured DRG neurons. Finally, we have used retrograde tract-tracing of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP) and NPY immunohistochemistry labeling methods to clarify the synaptic relationships between cuneothalamic projection neurons (CTNs) and NPY-LI terminals in the CN of CMNT rats. Ultrastructurally, the NPY-LI terminals made asymmetric axodendritic synaptic contact with HRP-labeling CTN dendrites. Taken together, these findings suggest that injury-induced NPY-LI fibers in the CN may originate from the injured DRG neurons via the median nerve primary afferent fibers, affect the excitability of CTNs, and involve neuropathic sensation following CMNT. Then, four weeks after CMNT when given electrical stimulation, c-Fos-like immunoreactive (c-Fos-LI) cells (mean density = 60.1 | en |
dc.description.provenance | Made available in DSpace on 2021-06-13T16:58:17Z (GMT). No. of bitstreams: 1 ntu-94-F88446008-1.pdf: 1874615 bytes, checksum: 968b6aaec18e237ee8a1f4408f4cd098 (MD5) Previous issue date: 2005 | en |
dc.description.tableofcontents | 目錄
誌謝與感言………………………………………………………………………...Ⅰ 中文摘要…………………………………………………………………………...Ⅱ 英文摘要…………………………………………………………………………...Ⅴ 引言…………………………………………………………………………………1 第一章 大白鼠正中神經截斷後存活不同時程,背根神經節中神經胜肽Y免疫反應神經元的表現情形 前言………………………………………………………………………14 材料與方法………………………………………………………………15 結果………………………………………………………………………18 討論………………………………………………………………………24 第二章 大白鼠正中神經截斷後,背根神經節中神經胜肽Y與神經損傷的關係 前言………………………………………………………………………26 材料與方法………………………………………………………………27 結果………………………………………………………………………28 討論………………………………………………………………………30 第三章 大白鼠正中神經截斷後,背根神經節與楔狀神經核內神經胜肽Y表現的相互關係 前言………………………………………………………………………33 材料與方法………………………………………………………………34 結果………………………………………………………………………35 討論………………………………………………………………………37 第四章 大白鼠正中神經截斷後,楔狀神經核內神經胜肽Y免疫反應神經纖維與楔狀丘腦接轉神經元的關係 前言………………………………………………………………………40 材料與方法………………………………………………………………41 結果………………………………………………………………………43 討論………………………………………………………………………45 第五章 大白鼠正中神經截斷後,楔狀神經核中神經胜肽Y與c-Fos表現的相互關係 前言………………………………………………………………………47 材料與方法………………………………………………………………48 結果………………………………………………………………………52 討論………………………………………………………………………55 第六章 不同藥劑處理,對正中神經損傷的大白鼠楔狀神經核中神經胜肽Y表現的影響 前言………………………………………………………………………58 材料與方法………………………………………………………………59 結果………………………………………………………………………61 討論………………………………………………………………………62 參考文獻……………………………………………………………………………66 圖表與說明…………………………………………………………………………77 | |
dc.language.iso | zh-TW | |
dc.title | 大白鼠正中神經截斷損傷後楔狀神經核及背根神經節中神經胜肽Y相關因子之研究 | zh_TW |
dc.title | The Related Factors of Neuropeptide Y in the Cuneate Nucleus and Dorsal Root Ganglion after Median Nerve Transection of Rats | en |
dc.type | Thesis | |
dc.date.schoolyear | 93-2 | |
dc.description.degree | 博士 | |
dc.contributor.oralexamcommittee | 溫振源(Chen-Yuan Wen),謝正勇(Jeng-Yung Shieh),謝松蒼(Sung-Tsang Hsieh),劉江川(Jiang-Chuan Liu),吳慶祥 | |
dc.subject.keyword | 神經神肽,Y,正中神經,楔狀神經核, | zh_TW |
dc.subject.keyword | neuropeptide Y,cuneate nucleus,median nerve, | en |
dc.relation.page | 145 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2005-04-13 | |
dc.contributor.author-college | 醫學院 | zh_TW |
dc.contributor.author-dept | 解剖學研究所 | zh_TW |
顯示於系所單位: | 解剖學暨細胞生物學科所 |
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