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研究生:黃薇如
研究生(外文):Wei-Ju Huang
論文名稱:第二型環氧化酶及前列腺素參與胰島素對雄性大鼠胃腸運動之效應
論文名稱(外文):Involvement of Cyclooxygenase 2 and Prostaglandin E2 in the Effects of Insulin on Gastriointestinal Motility in Male Rats
指導教授:王錫崗王錫崗引用關係董明倫董明倫引用關係
指導教授(外文):Paulus S. WangMing-Luen Doong
學位類別:博士
校院名稱:國立陽明大學
系所名稱:生理學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2009
畢業學年度:97
語文別:中文
論文頁數:255
中文關鍵詞:糖尿病胰島素第二型環氧化酶前列腺素腸胃道
外文關鍵詞:diabetesinsulincyclooxygenaseprostaglandin E2gastrointestinal tract
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臨床發現大部分無論第一型或是第二型糖尿病患者通常伴隨有胃腸運動不適的現象,往往在進食過後產生消化上的障礙,嚴重者甚至影響正常飲食及生理運作。在補充胰島素後可改善胃腸活動的不適。因此針對糖尿病症與所誘發胃腸之關聯性值得深入探究。本論文認為糖尿病腸胃道活動的異常與糖尿病誘發的發炎反應(Diabetic Inflammation)與神經性病變(Diabetic Neuropathy)有密切關聯。因此利用正常與糖尿病大鼠給予胰島素注射,並透過蕈膽鹼受體拮抗劑atropine進行胃腸運動實驗,同時利用Western Blot 觀察胰島素受體與發炎物質第二型環氧化酶COX-2表現,與其下游前列腺素PGE2 的分泌,並利用平滑肌肉收縮實驗觀測糖尿病對胃腸平滑肌肉的收縮效應。結果顯示在不同時期的糖尿病階段對於胃腸活動有不同的影響,初期胃腸活動受到抑制但是在後續則上升。同時觀察COX-2的改變發現初期有降低趨勢但是後期回升,而PGE2在糖尿病初期降低且平滑肌肉收縮亦弱化。在糖尿病初期給予胰島素補充後可回升胃腸活動,同時透過改變COX-2的表現與PGE2的生成影響平滑肌肉收縮。另一方面結果發現糖尿病大鼠透過蕈膽鹼受體使胃腸活動受到抑制,但是胰島素透過了第一與第三型蕈膽鹼受體逆轉了胃腸活動。本論文結論證實第一型糖尿病初期由於自體免疫開始破壞胰臟β細胞導致發炎反應開始進行,發炎反應開始時發炎指標第二型環氧化酶COX-2增加,此時由於胰臟β細胞尚未完全破壞導致初期糖尿病的假性正常現象。但是在發炎反應持續進行數小時後,即β細胞完全破壞造成血糖異常,胰島素作用異常,此時卻發生了第二型環氧化酶COX-2活性表現降低的現象,根據已知文獻了解COX-2於發炎反應後期有抗發炎的效應產生,同時推測由於糖尿病患者COX-2降低導致發炎現象加劇,進而促使更嚴重的發炎作用進行,再度刺激新的COX-2產生。糖尿病初期由於COX-2活性降低導致PGE2生成少,胃排空降低,小腸收縮遲緩,造成初期糖尿病有消化功能差,腹漲,心灼熱,便秘,等現象;後期由於發炎反應加重誘發新生COX-2再度生成,因此促使後期糖尿病患者反而產生胃排空增加,小腸收縮快導致腹瀉等併發症。胰島素的投入可透過與胰島素受體的結合進而改變COX-2的活性連帶影響的胃腸功能。此外在糖尿病神經性病變方面也解釋了由於初期糖尿病透過蕈膽鹼受體的破壞而導致外胃腸功能的遲緩而胰島素可透過M1與M3兩型受體改善糖尿病消化功能的異常。
Delayed gastric emptying in patients with both type 1 and type 2 diabetes mellitus (DM) occurs in approximately 50% of these patients. However, the role and the action mechanism of insulin on gastrointestinal (GI) motility are still unclear. The purpose of the present study was to investigate the involvement of cyclooxygenase-2 (COX-2) in diabetes inflammation, and muscarinic cholinergic receptor in diabetes neuropathy about the effects of insulin on GI motility in male rats. The normal and streptozotocin (STZ)-pretreated rats were injected intraperitoneally with or without insulin, atropine and specific muscarinic receptor antagonists before examination of measurement of gastric emptying, spontaneous contractile activity of smooth muscle strips, and prostaglandin E2 (PGE2) analysis. COX-2 expression and insulin receptor (IR) were analyzed by the technique of western blot. Our results showed that the GI motility was different between the early stage and the later period of DM. The COX-2 expression decreased after 1 week STZ-induced diabetes but was reversed several days later. Administration of insulin accelerated GI motility as well as the IR and COX-2 expression. In the early stage of DM, we observed the IR expression in GI and found that IR was changed under the insulin and DM treatment, and was also different between STZ-pretreated rats and hyperglycemic rats. Expression of COX-2 in stomach was decreased in DM rats but restored by insulin. The COX inhibitor, indomethacin, decreased the gastric emptying which was induced or reversed by insulin in normal and DM rats, respectively. PGE2 production in stomach corresponded to the COX-2 expression. The contraction of GI smooth muscle stimulated by PGE2 was increased in insulin-pretreated normal and DM rats. On the other hand, atropine interrupted the insulin effect on gastric emptying, and muscarnic M1/M3 receptor antagonists interrupted the insulin-reversed gastric emptying in normal and DM rats. We conclude that the side-effects of GI motility were associated with the COX-2 expression during diabetic inflammation. The GI smooth muscle contraction was affected via the COX-2 activation and PGE2 production. Besides, the delayed GI motility in early stage of diabetes was associated with the muscarinic cholinergic system in diabetic neuropathy. Insulin improved the delayed GI motility in the early stage of DM through the IR and COX-2 expression plus PGE2 production of COX-2 and PGE2 pathway in stomach and intestine as well as reversed the delayed gastric emptying via the nervous actions of muscarinic M1 and M3 receptors in DM rats.
目 錄
致謝…………………………………………………………………………5
中英文名詞對照表及英文縮寫……………………………………………11
藥物作用一覽表……………………………………………………………13
圖錄…………………………………………………………………………15
表錄…………………………………………………………………………17

主題一 第二型環氧化酶及前列腺素參與胰島素對雄性大鼠胃腸運動之效應
……………………………………………………………………19
中文摘要……………………………………………………………………21
英文摘要……………………………………………………………………23

第壹章、文獻回顧…………………………………………………………27
一、 概論…………………………………………………………………27
二、 糖尿病………………………………………………………………29
(一) 定義………………………………………………………………29
(二) 種類………………………………………………………………29
(三) 症狀………………………………………………………………31
(四) 診斷………………………………………………………………32
(五) 治療………………………………………………………………32
(六) 糖尿病對消化道的影響…………………………………………32
三、 胰島素………………………………………………………………32
(一) 來源………………………………………………………………32
(二) 構造………………………………………………………………32
(三) 功能………………………………………………………………33
(四) 胰島素與糖尿病的關聯…………………………………………33
四、 胃排空與胃腸收縮…………………………………………………33
(一) 腸胃道的構造與特性……………………………………………33
(二) 腸胃道與糖尿病之關聯…………………………………………35
五、 第一型糖尿病與發炎反應…………………………………………37
六、 環氧化酶……………………………………………………………38
(一) 構造與特性………………………………………………………38
(二) 分類………………………………………………………………38
(三) COX-2 與NSAID………………………………………………38
(四) COX-2 與消化道………………………………………………38
七、 前列腺素……………………………………………………………39
(一) 構造與分類………………………………………………………39
(二) PGE2……………………………………………………………41
八、 大麻受體……………………………………………………………41
(一) 構造與分類………………………………………………………41
(二) 大麻受體與糖尿病………………………………………………43
九、 第一型糖尿病與神經性病變(Diabetic neuropathy)………………43
十、 自主神經系統(autonomic nervous system) ………………………43
十一、 目的與假說……………………………………………………44
(一) 目的………………………………………………………………44
(二) 假說………………………………………………………………45
第貳章 材料與方法…………………………………………………………48
一、實驗動物………………………………………………………………48
二、大鼠糖尿病模式的建立………………………………………………48
三、胃排空及小腸通過率實驗……………………………………………48
(一) 原理………………………………………………………………48
(二) 方法………………………………………………………………48
四、血漿膽囊收縮素(CCK)濃度之檢測…………………………………50
(一) 原理………………………………………………………………50
(二) 方法………………………………………………………………50
五、小腸平滑肌組織之製備………………………………………………50
(一) 原理………………………………………………………………50
(二) 方法………………………………………………………………50
六、西方墨漬分析…………………………………………………………51
(一) 原理………………………………………………………………51
(二) 方法………………………………………………………………51
七、組織及血漿中前列腺素 prostagladin E1 (PGE1)及 E2 (PGE2) 之檢測:
利用酵素免疫分析法 (enzyme immunoassay, EIA) ………………53
(一) 原理與方法………………………………………………………53
八、統計分析………………………………………………………………53
第參章 論文第一部分 - 胰島素對於正常雄性大鼠胃腸運動之效應……55
一、 序言…………………………………………………………………57
二、 方法…………………………………………………………………57
三、 結果…………………………………………………………………61
四、 討論…………………………………………………………………63
第肆章 論文第二部分 - 胰島素對於糖尿病雄性大鼠胃腸運動之效應…69
一、 序言…………………………………………………………………71
二、 方法…………………………………………………………………72
三、 結果…………………………………………………………………76
四、 討論…………………………………………………………………78
第伍章 論文第三部分 - 胰島素對於糖尿病雄性大鼠胃腸道中胰島素受體表現之影響……………………………………87
一、 序言…………………………………………………………………88
二、 方法…………………………………………………………………89
三、 結果…………………………………………………………………90
四、 討論…………………………………………………………………90
第陸章 論文第四部分 - 胰島素對於糖尿病雄性大鼠胃腸道中第二型環氧化酶表現之影響………………………………97
一、 序言…………………………………………………………………98
二、 方法…………………………………………………………………99
三、 結果…………………………………………………………………99
四、 討論………………………………………………………………100
第柒章 論文第五部分 - 胰島素對於糖尿病雄性大鼠胃腸道及血漿中前列腺素分泌之影響………………………………107
一、 序言………………………………………………………………108
二、 方法………………………………………………………………108
三、 結果………………………………………………………………110
四、 討論………………………………………………………………111
第捌章 論文第六部分 -前列腺素對胰島素與糖尿病大鼠平滑肌收縮之效應………………………………………………115
一、 方法………………………………………………………………116
二、 結果與討論………………………………………………………116
第玖章 論文第七部分 - 胰島素對於糖尿病雄性大鼠胃腸道中大麻受體表現之影響………………………………………121
一、 序言………………………………………………………………122
二、 方法………………………………………………………………122
三、 結果………………………………………………………………123
四、 討論………………………………………………………………123
結論…………………………………………………………………………125

主題二 多巴胺與腎上腺素受體在安非他命抑制雄鼠胃腸活動作用中所扮演之角色……………………………………………………………129
中文摘要……………………………………………………………………131
英文摘要……………………………………………………………………133

第壹章、文獻回顧…………………………………………………………135
一、 概論………………………………………………………………137
二、 安非他命…………………………………………………………137
(一) 構造與分類……………………………………………………137
(二) 功能……………………………………………………………137
三、 多巴胺……………………………………………………………139
(一) 構造與分類……………………………………………………139
(二) 多巴胺受體之功能……………………………………………139
四、 腎上腺素受體……………………………………………………139
(一) 構造與分類……………………………………………………139
(二) 功能……………………………………………………………139
五、 目的與假說………………………………………………………140
第貳章、材料與方法………………………………………………………141
一、 實驗動物…………………………………………………………142
二、 胃排空及小腸通過率實驗………………………………………142
(一) 原理……………………………………………………………142
(二) 方法……………………………………………………………142
第參章 主題二論文第一部分–多巴胺受體對安非他命抑制雄鼠胃腸運動之調控………………………………145
一、 序言………………………………………………………………146
二、 方法………………………………………………………………146
三、 結果………………………………………………………………147
四、 討論………………………………………………………………148
第肆章 主題二論文第二部分–腎上腺素受體對安非他命抑制雄鼠胃腸運動之調控……………………………153
一、 序言………………………………………………………………154
二、 方法………………………………………………………………154
三、 結果………………………………………………………………156
四、 討論………………………………………………………………157
結論…………………………………………………………………………165
參考文獻……………………………………………………………………169
附錄…………………………………………………………………………185
個人履歷…………………………………………………………………187
已接受發表及投稿中之論文……………………………………………190
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