脊髓損傷、急性疼痛、和嗎啡疼痛耐受性都是臨床上重要的課題。初次傷害引起脊髓型態損傷和生化變化，續發毀滅性的二次傷害，最後摧毀了中樞神經系統、造成神經退化。週邊組織的損傷合併著疼痛接受體的敏感性以及相繼而來發生的中樞神經元的興奮，這些被稱為中樞的敏感性。而疼痛接受體的敏感性和中樞的敏感性被認為是引發原發性和次發性過痛的原因。目前最有效之止痛藥物仍為嗎啡類止痛劑，長期給予這些藥物會產生耐受性，耐受性是指藥物效力的逐次減弱，同時產生後必須將劑量加大才可達到一定的止痛效果。嗎啡耐受性分子生物學的機轉仍舊不明。

近來發現calpain-Cdk5(cyclin-dependent kinase-5)-p35路徑的調控和神經保護、急性疼痛、嗎啡止痛有關。在本論文中，我們以calapain抑制劑-MDL28170和Cdk5抑制劑-roscovitine治療大白鼠脊髓半切除損傷、福馬林引起的急性疼痛、和慢性嗎啡耐受性。結果，抑制calpain-Cdk5-p35路徑可以保護大白鼠脊髓半切除損傷、並避免之後的神經退化，抑制福馬林引起的疼痛縮腳反應、並調控DARPP-32(dopamine and c-AMP regulated phosphoprotein, MW=32 kDa)的磷酸化，將已右移的嗎啡止痛劑量-反應曲線向左反轉、並調降已上昇的ED50(有效劑量的50％)。結論，抑制calpain-Cdk5-p35路徑對於脊髓損傷、急性疼痛、和嗎啡耐受性治療有效，並具有臨床應用的價值。

Spinal cord injury, acute pain, and morphine tolerance are important issues in the clinical practice. A primary injury to the spinal cord causes both morphological and biochemical changes with initiation of the devastating secondary pathophysiological pathways that ultimately destroy CNS cells and cause degeneration of nerve fibers. Tissue injury is associated with sensitization of nociceptors and subsequent changes in the excitability of central neurons, known as central sensitization. Nociceptor sensitization and central sensitization are believed to underlie the development of primary and secondary hyperalgesia, respectively. The most efficacious drugs used to relieve pain are the opioid analgesics. Chronic administration leads to the development of tolerance. Tolerance is manifested as a decreased potency of the drug, so that progressively larger doses must be administered to achieve a given level of analgesia. The processes underlying opioid tolerance still need to be elucidated.

Recently, it is found calpain-Cdk5 (cyclin-dependent kinase-5)-p35 pathway modulation implicated in neuroprotection, acute nociceptive response, and morphine analgesia. In this thesis, we evaluate calpain inhibitor-MDL28170 and Cdk5 inhibitor-roscovitine against rat spinal cord hemisection, formalin-induced acute nociceptive responses, and chronic morphine tolerance. We found calpain-Cdk5-p35 pathway inhibition could protect spinal cord hemisection and subsequent neurodegeneration, inhibit formalin-induced flinch response involving DARPP-32 (dopamine and c-AMP regulated phosphoprotein, MW=32 kDa) phosphorylation, and reverse right shifted morphine dose-response curve with upregulated ED50 (50% of effective dose) reduction. Taken together, calpain-Cdk5-p35 pathway inhibition is useful in the management of spinal cord injury, acute inflammatory pain, and attenuate morphine tolerance development with further clinical application.

目錄
序 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ I
序章 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ II
目錄 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ IV
中文摘要 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ VI
Abstract ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ VIII

第一章 緒論及文獻回顧 ˙˙˙˙˙˙˙˙˙˙˙ p1
1-1 細胞死亡:壞死和細胞凋亡˙˙˙˙˙˙˙˙ p2
1-2 Calpain ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p3
1-3 Calpain的活化機轉˙˙˙˙˙˙˙˙˙˙˙ p3
1-4 Calpain抑制劑 ˙˙˙˙˙˙˙˙˙˙˙˙ p4
1-5 Calpain的受質 ˙˙˙˙˙˙˙˙˙˙˙˙ p5
1-6 Calpain抑制劑應用於動物模式˙˙˙˙˙˙ p8
1-7 Calpain和脊髓損傷˙˙˙˙˙˙˙˙˙˙˙ p8
1-8 抑制Calpain活性以治療脊髓損傷 ˙˙˙˙˙p9
1-9 Calpain-Cdk5-p35路徑˙˙˙˙˙˙˙˙˙˙p10
1-10 Alzheimer Disease和Tau˙˙˙˙˙˙˙˙˙p11
1-11 Cyclin-dependent kinase-5 ˙˙˙˙˙˙˙p13
1-12 研究的動機以及目的 ˙˙˙˙˙˙˙˙˙˙ p14

第二章 大白鼠椎管內注射roscovitine
抑制Cdk5的活性並減緩由福馬林
引起的疼痛縮腳反應 ˙˙˙˙˙˙˙˙˙ p16
2-1 摘要 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p17
2-2 緒論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p17
2-3 材料與方法 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p19
2-4 結果 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p21
2-5 討論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p22

第三章 大白鼠椎管內注射roscovitine
延緩嗎啡疼痛耐受性的產生 ˙˙˙˙˙˙ p28

3-1摘要 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p29
3-2 緒論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p29
3-3 方法和材料 ˙˙˙˙˙˙˙˙˙˙˙˙˙ p31
3-4 結果 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p33
3-5 討論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p34

第四章 Calpain抑制劑抑制p35-p25-Cdk5
活化, 避免tau過度磷酸化及保護
大白鼠脊髓半切除的神經損傷 ˙˙˙˙˙ p39

4-1 摘要 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p40
4-2 緒論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p41
4-3 材料與方法˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p43
4-4 結果 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p48
4-5 討論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p50

第五章 綜合討論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙p74
5-1 結論 ˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙ p75
5-2 本論文之重要性 ˙˙˙˙˙˙˙˙˙˙˙˙ p79
5-3 未來研究方向 ˙˙˙˙˙˙˙˙˙˙˙˙˙ p80

參考文獻˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙˙p82

已發表論文一覽表˙˙˙˙˙˙˙˙˙˙˙˙˙˙p99

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