博碩士論文 982204019 詳細資訊




以作者查詢圖書館館藏 以作者查詢臺灣博碩士 以作者查詢全國書目 勘誤回報 、線上人數:22 、訪客IP:18.116.13.171
姓名 廖元邦(Benjamin Yuan-Bang Liau)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 蛋白磷酸激酶A 與蛋白磷酸激酶C epsilon 參與在 酸以及溶血磷脂質引起的疼痛敏感現象
(Protein Kinase A and Protein Kinase C epsilon Are Involved in Acid- and Lysophospholipids-induced Hyperalgesia)
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摘要(中) 中文摘要
痛覺是ㄧ種當身體受到環境中的有害刺激時,所產生的一種不舒服的重要感
覺,也是警告我們身體遭受到傷害,必須規避或治療的一種保護機制。
前人研究發現在海藻醣引發的預發炎模式中,蛋白激酶A 與Cε 可能扮演在
急性疼痛轉為慢性疼痛的關鍵角色。此外,組織酸化被認為是造成疼痛的主要原
因之一,但目前還不清楚的是因組織酸化而引起疼痛是透過怎麼樣的下游機制。
此外細胞內的溶血磷脂質是否能引發痛覺,會參與在哪類痛覺的機制中,也仍然
尚未被釐清。
我探究的方式是在小鼠的後肢腳掌皮下注射酸性緩衝溶液與三種溶血磷脂
質,並且分別在各組實驗裡另注射不同訊號傳遞路徑的抑制劑。注射後在不同的
時間點分別用馮法瑞尼龍測量絲以及哈格里夫斯熱感測試儀來做測試,觀察機械
性疼痛和熱感刺激疼痛是否被下游路徑的抑制劑所抑制,以及抑制的程度與時間
區段。實驗發現注射酸以及三類溶血磷脂質都造成機械性與熱感痛覺敏感性增強
的現象並持續超過一天。另外發現蛋白激酶A 的抑制劑、AC 之阻斷劑會在疼痛
敏感性上升的初期有抑制作用;蛋白激酶Cε 的抑制劑、Gi 之抑制劑、PLC 之阻
斷劑則會在疼痛敏感性上升的中期以後才有抑制作用。有關痛覺的離子通道方面
的ASICs 之阻斷劑、TRP 之阻斷劑,也能抑制機械性和熱感痛覺敏感性升高的
現象。期許在更多的痛覺機制被釐清之後,能被應用在研發緩和痛覺,副作用更
少的藥物上。
摘要(英) Abstract
Pain is a complex experience that involves the transduction of noxious
environmental stimuli. The ability to detect noxious stimuli is essential to an
organism’s survival and wellbeing.
Some studies have demonstrate that in carrageenan-induced priming state protein
kinase A and Cε play critical roles in the transition from acute pain to chronic pain. A
decrease in tissue pH has been observed following inflammation, hematomas, and
isometric exercise. A positive correlation exists between pain and local acidity.
However, the downstream mechanism of proton-induced pain remains unclear.
Furthermore, the regulatory pathway between lysophospholipids and pain has not
been investigated completely.
To answer these questions, I used mechanical and thermal tests in different time
period to examine the hyperalgesia induced by acidic solution, psychosine,
lysphosphatidylcholine and sphingosylphosphatidylcholine. In addition, protein
kinase A inhibitor and adenylyl cyclase blocker inhibit hyperalgesia in the early stage,
but protein kinase Cε inhibitor, Gi inhibitor and phospholipase C blocker inhibit
hyperalgesia in the middle and late stage. Transient receptor potential family blocker
and acid-sensing ion channel blocker also inhibit acid-induced mechanical and
thermal hyperalgesia. I wish these studies can be used in pain curation in the future.
關鍵字(中) ★ 痛覺
★ 蛋白磷酸激酶
★ 疼痛敏感
關鍵字(英) ★ pain
★ protein kinase
★ hyperalgesia
論文目次 目錄
第一章 緒論 ………………………………………………………………………. 1
1.1 痛覺 ………………………………………………………………………….. 2
1.2 痛覺的訊息傳導途徑 ……………………………………………………….. 3
1.3發炎物質引起痛覺過敏現象 ………………………………………………... 5
1.4 疼痛與組織酸化 …………………………………………………………….. 6
1.5 與氫離子相關的離子通道 ………………………………………………….. 7
1.5.1辣椒素受體 ……………………………………………………………… 8
1.5.2 酸敏感離子通道家族 …………………………………………………... 8
1.6 與痛覺訊號傳遞相關的溶血磷脂質 ……………………………………….. 9
1.6.1 脫脂磷酸脂膽鹼 ………………………………………………………. 10
1.6.2 神經磷醯膽鹼 …………………………………………………………. 11
1.6.3 硝氨醇半乳糖 …………………………………………………………. 12
1.7 疼痛傳遞可能經由的GPCR路徑 ………………………………………… 13
1.7.1 G 蛋白偶合受體 ………………………………………………………. 13
1.7.2 G蛋白偶合受體下游反應路徑 …………………………………………14
1.7.3 蛋白磷酸化酶 A ………………………………………………………. 15
1.7.4 蛋白磷酸化酶 C ………………………………………………………. 15
1.7.5 蛋白磷酸化酶 A 和蛋白磷酸化酶 Cε 參與的疼痛反應 ………… 16
1.8 研究動機與目的 …………………………………………………………… 17
第二章 材料與方法 ………………………………………………………………19
2.1 實驗材料 …………………………………………………………………… 20
2.1.1 實驗動物 ……………………………………………………………… 20
2.1.2 實驗藥品 ……………………………………………………………… 20
2.2 實驗方法 …………………………………………………………………… 23
2.2.1 動物疼痛行為實驗 …………………………………………………… 23
2.2.1.1 小鼠腳掌皮下注射 ……………………………………………… 23
2.2.1.2 機械性疼痛行為實驗 …………………………………………… 23
2.2.1.3 熱疼痛行為實驗 ………………………………………………… 25
2.3 統計分析 …………………………………………………………………… 26
第三章 結果 …………………………………………………………………….. 28
3.1 脫脂磷酸脂膽鹼、神經磷醯膽鹼與硝氨醇半乳糖的注射,會提高對於機械性疼動的痛覺敏感性 …………………………………………………………….. 29
3.2 脫脂磷酸脂膽鹼、神經磷醯膽鹼與硝氨醇半乳糖的注射,會提高對於熱感疼動的痛覺敏感性 ……………………………………………………………….. 31
3.3 PKA的抑制劑能在LPC注射後的0-90分鐘內抑制LPC造成的機械性與熱感疼痛閾值降低的現象;PKCε的抑制劑的抑制作用時間則是在LPC注射後的120-240分鐘間 …………………………………………………………………… 33
3.4 Gi的抑制劑能夠降低在注射LPC、SPC、psychosine的60-240分鐘間,所造成的機械性疼痛與熱感疼痛閾值的降低現象 …………………………………35
3.5 注射pH低的溶液會增加機械性疼痛和熱感疼痛的敏感性。隨著pH值的降低與昇高,疼痛敏感性也會隨之增強與減弱 …………………………………37
3.6 PKA的抑制劑可以在注射酸後的180分鐘內,抑制注射酸造成的機械性疼痛敏感閾值降低的現象 ……………………………………………………………39
3.7 PKCε的抑制劑可以在注射酸後的180-480分鐘內,抑制注射酸造成的機械性疼痛敏感閾值降低的現象 ………………………………………………………41
3.8 PKA的抑制劑可以在注射酸後的120分鐘內,抑制注射酸造成的熱感疼痛敏感閾值降低的現象;PKCε的抑制劑則可以在注射酸後的240-480分鐘內,抑制注射酸造成的熱感疼痛敏感閾值降低的現象 …………………………………43
3.9 PLC的阻斷劑可以在注射酸後的60-240分鐘內,抑制酸造成的機械性疼痛敏感閾值降低的現象;Gi的抑制劑對酸造成的機械性疼痛敏感與熱感疼痛敏感閾值降低的抑制時間為注射後的60-240分鐘內;AC的阻斷劑對酸造成的機械性疼痛敏感閾值降低的抑制時間則為注射酸後的60-120分鐘內 …………………45
3.10 ASICs的阻斷劑會在注射酸後的240分鐘內抑制酸造成的機械型疼痛敏感閾值降低的現象;TRP的阻斷劑對酸造成的機械性疼痛敏感與熱感疼痛敏感閾值降低的抑制時間則是在注射酸後的90-180分鐘之間 ……………………………47
第四章 討論 ………………………………………………………………………50
4.1 注射 LPC、SPC 與 psychosine 造成之機械性與熱感痛覺敏感現象 … 51
4.2 PKA 和 PKC 的抑制劑對 LPC 造成之疼痛敏感的抑制 ……………… 53
4.3 Gi 的抑制劑對 LPC、SPC 和 psychosine 造成之疼痛敏感的抑制 …… 54
4.4 酸造成之機械性與熱感痛覺敏感現象 …………………………………… 55
4.5 PKA 和 PKC 的抑制劑對酸造成之疼痛敏感的抑制 ……………………57
4.6 PLC、Gi 和 AC 的抑制劑對酸造成之疼痛敏感的抑制 ………………… 59
4.7 ASICs 和 TRP 的阻斷劑對酸造成之疼痛敏感的抑制 ………………… 60
第五章 參考文獻 …………………………………………………………………62
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指導教授 孫維欣(Wei-Hsin Sun) 審核日期 2013-7-17
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