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研究生:施春兆
研究生(外文):Chuen-Chao Shi
論文名稱:駱駝蓬及穿心蓮藥效成分舒張離體血管及氣管平滑肌之探討
論文名稱(外文):Study on the relaxation effects of the principles of Peganum harmala L and Andrographis paniculata on vascular and tracheal smooth muscles in vitro
指導教授:陳介甫陳介甫引用關係廖志飛廖志飛引用關係
指導教授(外文):Chieh-Fu ChenJyh-Fei Liao
學位類別:博士
校院名稱:國立陽明大學
系所名稱:藥理學研究所
學門:醫藥衛生學門
學類:藥學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:192
中文關鍵詞:駱駝蓬穿心蓮血管舒張一氧化氮內皮細胞
外文關鍵詞:peganum harmalaandrographis paniculatavasorelaxantnitric oxideendothelium
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延伸先前對駱駝蓬與穿心蓮抽提物之藥理研究,本論文主要是探討四種駱駝蓬生物鹼(harmine、harman、harmaline和harmalol)及三種穿心蓮成分(穿心蓮甲素、穿心蓮乙素、穿心蓮丙素)之降血壓、離體血管舒張、抗血小板凝集及離體氣管平滑肌舒張作用,並評估其對相關接受器之親和力。在駱駝蓬生物鹼之研究方面,發現靜脈給予此四種生物鹼(1-10 mg/kg)對於麻醉大白鼠具有劑量相關性的短暫降血壓及減緩心跳作用,而此作用可被一氧化氮(NO)合成抑制劑L-NNA (2.5 mg/kg, i.v.)預處理所抑制,顯示NO參與它們的心臟血管作用。這四種生物鹼皆以濃度相關地舒張由phenylephrine (PE),5-HT及KCl引致大鼠離體胸主動脈之收縮,其中harmalol之血管舒張作用是非內皮依賴性,而其餘三種生物鹼之作用是部份內皮依賴性。其作用在內皮細胞可使內鈣釋放或/和外鈣內流而增加[Ca2+]i 導致NO合成與釋放;其作用在血管平滑肌可同時抑制無外鈣下PE引致之瞬間性血管收縮及再加入鈣離子引致之持續性收縮,然此拮抗PE與5-HT引致之血管收縮是屬於非競爭性,另以受體結合實驗顯示,在相似之濃度下對於大鼠心臟之α1-adrenoceptor及小鼠大腦皮質之5-HT2接受器均具有親和力。對於人類和兔子之血小板,harmine、harman和harmaline均可抑制不同凝集刺激劑(如:AA與TXA2)引致之血小板凝集。此抑制arachidonic acid (AA)引致之血小板凝集與抑制AA引致之血小板[Ca2+]i增加可能與thromboxane A2 (TXA2)形成有關。對於天竺鼠離體氣管條,這四種生物鹼亦以濃度相關地舒張因carbachol、histamine及KCl引致之氣管收縮。然此抑制carbachol所引起氣管收縮可能部份經由活化β-adrenoceptor外,直接以競爭性拮抗muscarinic 接受器所致。此外在穿心蓮成分之研究方面,發現穿心蓮甲素和穿心蓮乙素皆以濃度相關地舒張由PE,5-HT及KCl引致大鼠離體胸主動脈之收縮,其中穿心蓮甲素之血管舒張作用部份是內皮依賴性,其作用在內皮細胞可使外鈣內流而增加[Ca2+]i 導致NO合成與釋放,而穿心蓮甲素與穿心蓮乙素作用在血管平滑肌也以非競爭性的拮抗PE或5-HT引致之氣管收縮。三種穿心蓮成分均可同時抑制無外鈣下PE引致之瞬間性收縮及再加入鈣離子引致之持續性收縮。穿心蓮乙素對大白鼠心臟之α1-adrenoceptor有微弱親和力,因此推測其血管舒張作用可能不是作用在接受器層面。對於人類和兔子之血小板,三種穿心蓮成分對於不同凝集刺激劑(如:AA與TXA2)有不同程度之抑制作用,其中以對抗collagen之效力最強。另穿心蓮甲素抑制AA引致之血小板凝集與抑制AA引致之血小板[Ca2+]i增加可能與TXA2形成有關。對於天竺鼠離體氣管條,穿心蓮甲素可舒張carbachol、histamine及KCl引致之收縮,而穿心蓮乙素及丙素僅對histamine引起收縮有舒張作用,而三種成分拮抗histamine引致之收縮是非競爭性。接受器結合實驗顯示穿心蓮丙素對於天竺鼠肺臟histamine H1接受器之親和力與其抑制histamine引起氣管收縮之作用效力相當,而穿心蓮甲素只有微弱的親和力,穿心蓮乙素則明顯無作用。綜合上述,本論文闡述四種駱駝蓬生物鹼及三種穿心蓮成分之血管舒張、抗血小板凝集及氣管舒張作用並部份解開其可能之作用機轉。
Following previous studies on the pharmacological effects of Peganum harmala and Andrographis paniculata extracts, the present study mainly examined the in vivo cardiovascular, in vitro vasorelaxant, antiplatelet, and tracheal spasmolytic effects of 4 psychological active principles (harmine, harman, harmaline, and harmalol) from the seeds of Peganum harmala and 3 active principles (deoxyandrographolide, andrographolide, and neoandrographolide) from the leafs of Andrographis paniculata.
The following results were obtained in the study on harmala alkaloids. In pentobarbital-anesthetized rats, harmine, harman, harmaline, and harmalol (1-10 mg/kg, i.v.) dose-dependently produced transient hypotension and bradycardia, which were attenuated by a nitric oxide (NO) synthase inhibitor NG-nitro-L-arginine (2.5 mg/kg, i.v.) pretreatment, indicating the involvement of NO in their cardiovascular effects. In isolated rat endothelium-intact thoracic aortic rings, these 4 harmala alkaloids concentration-dependently relaxed phenylepherine (PE)-, 5-HT-, and KCl-induced contractions. The vasorelaxant effects of harmine, harman and harmaline were partially endothelium-dependent, but harmalol was endothelium-independent. In cultured rat aortic endothelial cells, harmine, harman, and harmaline caused an increase in [Ca2+]i by inducing the influx of external Ca2+ and/or increasing the release of internal Ca2+, which in turn increased the synthesis and release of NO. In vascular smooth muscle, pretreatment with these 4 alkaloids reduced both the phasic contraction induced by PE in the absence of external Ca2+ and the tonic contraction induced by reintroduction of 2.5 mM CaCl2, and inhibited PE- and 5-HT-induced contractions in a noncompetitive manner. However, receptor binding assays indicated that harmine, harman, harmaline and harmalol interacted with the rat cardiac a1-adrenoceptors and mouse cerebral cortex 5-HT2 receptors in the same concentrations. In human and rabbit platelets, harmine, harman, and harmaline inhibited platelet aggregations induced by different aggregation inducers. The inhibition of arachidonic acid (AA)-induced aggregation was associated with the inhibitory effects on AA-induced increases in [Ca2+]i and in the synthesis of thromboxan A2 (TXA2). In isolated guinea-pig tracheal preparations, these 4 alkaloids also concentration-dependently relaxed the contractions induced by carbachol, histamine, and KCl. The relaxant effect of harmine, harman, or harmaline for carbachol-induced contractions involved the activation of b-adrenoceptors and the competitive antagonism on muscarinic acetylcholine receptors.
The following results were obtained in the study on the principles of Andrographis paniculata. In isolated rat endothelium-intact thoracic aortic rings, deoxyandrographolide and andrographolide concentration-dependently relaxed PE-, 5-HT-, and KCl-induced contractions. The vasorelaxant effect of deoxyandrographolide was partially endothelium-dependent. In cultured rat aortic endothelial cells, deoxyandrographolide caused an increase in [Ca2+]i by inducing the influx of external Ca2+, which in turn increased the synthesis and release of NO. In vascular smooth muscle, deoxyandrographolide and andrographolide inhibited PE- and 5-HT-induced contractions in a noncompetitive manner and pretreatment with these 3 principles reduced both the phasic contraction induced by PE in the absence of external Ca2+ and the tonic contraction induced by reintroduction of 2.5 mM CaCl2. However, receptor binding assays indicated that only andrographolide had a low affinity for rat cardiac α1-adrenoceptors, suggesting that the vasorelaxant effects of these 3 principles did not acting on the receptor level. In human and rabbit platelets, these 3 principles inhibited platelet aggregations induced by different aggregation inducers, with the action on collagen-induced aggregation being the most potent. Inhibition of AA-induced aggregation by deoxyandrographolide was associated with its inhibitory action on AA-induced increases in [Ca2+]i and in the synthesis of TXA2. In guinea-pig tracheal preparations, deoxyandrographolide relaxed carbachol-, histamine-, and KCl-induced contractions, but andrographolide and neoandrographolide only relaxed histamine-induced contractions. The effects of these 3 principles on histamine-induced contractions were noncompetitive. Receptor binding assays indicated that neoandrographolide interacted with guinea-pig lung histamine H1 receptor with a Ki value comparable with its potency to inhibit histamine-induced contractions; in contrast, deoxyandrographolide had low affinity and andrographolide was without significant interaction.
In summary, the present study demonstrated the vasorelaxant, antiplatelet, and tracheal spasmolytic effects of 4 harmala alkaloids and 3 active principles of Andrographis paniculata and also partially resolved their possible underlying mechanisms of action.
封面
謝誌
縮寫表
中文摘要
英文摘要
第一章 緒論
第一節 駱駝蓬生物鹼簡介
第二節 穿心蓮之簡介
第三節 研究目的
第四節 文獻回顧
第二章 實驗材料與方法
第一節 實驗材料之來源
第二節 降血壓活體實驗
第三節 離體血管實驗
第四節 離體血管內皮細胞之實驗
第五節 血小板凝集試驗
第六節 離體天竺鼠氣管平滑肌之張力實驗
第七節 藥物接受器結合實驗
第八節 統計分析
第三章 實驗結果
第一節 駱駝蓬生物鹼之藥理作用
一、對麻醉大白鼠的血壓與心跳之影響
二、對大白鼠胸主動脈環之舒張作用
三、對培養血管內皮細胞之作用
四、對血小板凝集作用之影響
五、對離體天竺鼠氣管平滑肌之舒張作用
六、對不同接受器之親和力
第二節 穿心蓮成分之藥理作用
一、對大白鼠胸主動脈環之舒張作用
二、對培養血管內皮細胞之作用
三、對血小板凝集作用之影響
四、對離體天竺鼠氣管平滑肌之舒張作用
五、對不同接受器之親和力
第四章 討論
第一節 駱駝蓬生物鹼之藥理作用
第二節 穿心蓮成分之藥理作用
第五章 結論
第六章 參考文獻
第七章 附錄
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