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研究生:鄭崴文
研究生(外文):Wei-Wen Cheng
論文名稱:柳杉葉部精油對小鼠中樞神經系統影響及口服急毒性試驗評估
論文名稱(外文):Effect of the Essential Oil from Cryptomeria japonica Leaves on the Central Nervous System and Acute Oral Toxicity Evaluation in Mice
指導教授:王升陽
學位類別:碩士
校院名稱:國立中興大學
系所名稱:森林學系所
學門:農業科學學門
學類:林業學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:86
中文關鍵詞:柳杉精油神經藥理學活性芬多精口服急毒性
外文關鍵詞:Cryptomeria japonicaessential oilneuropharmacological activitiesphytoncideacute oral toxicity test
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森林浴(green shower;shinrin-yoku)對人類的健康具有許多之益處,本研究乃針對柳杉林內所釋放出來的森林芬多精,利用微固相萃取法以及氣相層析儀進行組成分分析。分析結果顯示,柳杉林內所釋放出之芬多精之主要成分為α-pinene (19.35%)、β-myrcene (16.98%)、d-limonene (15.21%)以及γ-muurolene (7.42%);精油之主要成分為elemol (18.22%)、16-kaurene (11.63%)、3-carene (9.66%)、sabinene (9.37%)、4-terpineol (9.06%)、α-eudesmol (5.70%)、α-pinene (5.62%)以及d-limonene (5.26%)。此外,本研究續利用數種動物行為模型來評估柳杉葉部精油及其重要之單萜類,d-limonene,對小鼠中樞神經系統的作用與影響。結果發現柳杉葉部精油具有明顯的延長戊巴比妥誘導小鼠之睡眠持續時間;在高腳十字迷宮試驗與冰醋酸扭體試驗中也顯示,柳杉葉部精油與d-limonene皆具有抗焦慮及鎮痛之功用;並且可平緩咖啡因誘導的興奮情形,以及具有保護小鼠受到戊四唑驚厥之傷害。
為探討柳杉葉部精油使用之安全性,本研究以ICR品系小鼠進行口服急毒性試驗。試驗結果顯示,柳杉葉部精油對小鼠口服急毒性之半致死劑量(LD50)為9.83 g/kg B.W.,且經精油處理小鼠之腦、心、肝、腎、脾及胸腺等重要臟器,均無因試驗物質引起肉眼及組織病理變化;血液學檢測中之紅血球數、血紅素、血球容積比、平均血紅素濃度及血小板等相較於對照組均無影響;血清生化檢測中之麩氨酸氨基轉氨酶、麩氨酸丙酸轉氨酶與肌氨酸等相較於對照組亦沒有影響(p>0.05)。
Forest-air bathing and walking (green shower, shinrin-yoku) is beneficial on human health. In this study the phytoncide released from Cryptomeria japonica plantation forestry was collected by using solid-phase micro extraction (SPME) and followed by GC/MS analysis, the composition of phytoncide was characterized. The main compositions of phytoncide were α-pinene (19.35%), β-myrcene (16.98%), d-limonene (15.21%) and γ-muurolene (7.42%). On the other hand, the essential oil of C. japonica was obtained by water distillation and anglicized by GC/MS, its main composition were elemol (18.22%)、16-kaurene (11.63%)、3-carene (9.66%)、sabinene (9.37%)、4-terpineol (9.06%)、α-eudesmol (5.70%)、α-pinene (5.62%), and d-limonene (5.26%). Furthermore, the neuropharmacological activity of the essential oils from leaves of C. japonica (ECJ) was evaluated by several animal behavior tests. ECJ could prolong the sleeping phase of ICR mice in pentobarbital-induced sleeping time model. Besides, both EJC and one of its monoterpene, d-limonene possessed the potent anxiolytic and analgestic.activities based on the results obtained from elevated plus-maze and writhing tests. The formulation also antagonized the behavioral effects of CNS-stimulant drug caffeine, and protected mice from PTZ induced convulsions.
To evaluate the safety of EJC, the acute oral toxicity test in ICR mice was conduct in this study. The LD50 of EJC was estimated 9.83 g/kg body weight for mice. No significant lesions of brain, heart, liver, kidney, spleen and thymus were found at gross and histopathological observation in the treated mice. Hematological parameters included of RBC, HGB, HCT, MCH, and PLT values were as normal as control. Besides, serum biochemistry changes in liver and renal function included of ALT, AST and creatinine values had no significant difference (p > 0.05) with control.
目錄 …………………………………………………………………………Ⅰ
表目次 ………………………………………………………………………Ⅴ
圖目次 ………………………………………………………………………Ⅶ
摘要…………………………………………………………………………1
Abstract……………………………………………………………………… 2
壹、前言………………………………………………………………………4
貳、文獻回顧…………………………………………………………………7
一、文明病的產生與治療藥物………………………………………………7
(一) 胺基丁酸(GABA)………………………………………………………8
(二) 焦慮解除劑與安眠藥…………………………………………………10
1. Benzodiazepines…………………………………………………………11
2. Barbiturates………………………………………………………………12
二、 動物行為研究模式介紹………………………………………………13
(一) 動物自發行為之觀察……………………………………………………13
(二) 動物協調性之觀察……………………………………………………14
(三) 動物安眠試驗之觀察…………………………………………………14
(四) 動物驚厥模型…………………………………………………………14
(五) 動物疼痛模型…………………………………………………………15
(六) 動物焦慮模型…………………………………………………………15
(七)動物憂鬱模型 ……………………………………………………………15
(八) 動物行為紀錄分析系統 ………………………………………………16
三、柳杉抽出成分之研究 …………………………………………………18
(一) 萜類……………………………………………………………………19
1. 單萜類……………………………………………………………………20
2. 倍半萜類…………………………………………………………………20
3. 二萜纇……………………………………………………………………21
4. 三萜類……………………………………………………………………21
(二) 聚酚類……………………………………………………………………21
1. 木酚素……………………………………………………………………22
2. 類木酚素…………………………………………………………………22
3. 黃酮類……………………………………………………………………22
4. 其他化合物………………………………………………………………23
五、林木精油活性研究法及柳杉精油成分 …………………………………24
(一) 植物的揮發成分………………………………………………………24
(二) 精油的利用……………………………………………………………25
(三) 氣相層析………………………………………………………………26
(四) 柳杉精油之成分………………………………………………………29
1. 心材中萜類含量…………………………………………………………29
2. 邊材中萜類含量…………………………………………………………29
3. 葉部中萜類含量…………………………………………………………29
4. 樹皮中萜類含量…………………………………………………………29
四、 具生物活性之柳杉抽出成份…………………………………………33
(一) 抗真菌活性……………………………………………………………33
(二) 抗蟲活性………………………………………………………………34
(三) 抗植物病原菌活性……………………………………………………35
(四) 抗螨活性………………………………………………………………35
(五) 抗發炎和抗氧化活性…………………………………………………36
(六) 細胞毒殺活性…………………………………………………………36
(七) 抗細菌活性……………………………………………………………36
(八) 其他活性………………………………………………………………36
參、材料與方法………………………………………………………………37
一、試驗材料 ………………………………………………………………37
(一) 柳杉……………………………………………………………………37
(二) 實驗動物………………………………………………………………37
二、柳杉葉部精油與柳杉林芬多精 …………………………………………37
(一) 柳杉葉部精油之萃取…………………………………………………37
(二) 柳杉含水率及其精油之收率…………………………………………38
(三) 野外採樣………………………………………………………………38
(四) 氣相層析分析…………………………………………………………39
三、各種動物模型試驗方法 …………………………………………………39
(一) 戊巴比妥誘導催眠小鼠試驗(安眠的動物模型) ………………………39
(二) 強迫游泳試驗(憂鬱動物模型) …………………………………………40
(三) 高腳十字迷宮(焦慮動物模型) …………………………………………41
(四) 開放場地測試(open field test)…………………………………………42
(五) 咖啡因誘導小鼠興奮試驗……………………………………………43
(六) 冰醋酸扭體法(鎮痛動物模型) …………………………………………43
(七) 熱刺激甩尾法(鎮痛動物模型) …………………………………………43
(八) 戊四唑對小鼠驚厥之影響(驚厥試驗) …………………………………44
(九) 動物行為分析…………………………………………………………44
四、柳杉精油口服急毒性試驗 ………………………………………………45
(一) 口服急毒性之試驗……………………………………………………45
(二) 血液學檢查……………………………………………………………45
(三) 血清生化檢查…………………………………………………………46
(四) 臟器病理檢查…………………………………………………………46
(五) 結果分析………………………………………………………………46
五、 統計分析………………………………………………………………46
肆、結果與討論 ……………………………………………………………47
一、柳杉精油成分含量分析 …………………………………………………47
二、野外採樣樣品分析 ………………………………………………………49
三、柳杉葉部精油與d-limonene對小鼠中樞神經之影響 …………………51
(一) 柳杉葉部精油物對小鼠安眠之影響…………………………………51
(二) 柳杉葉部精油對強迫游泳試驗小鼠之影響…………………………52
(三) 柳杉葉部精油對小鼠抗焦慮之影響(高腳十字迷宮試驗)………………53
(四) 柳杉葉部精油對小鼠的自發行為影響(開放場地測試) ……………… 56
(五) 柳杉葉部精油對咖啡因誘導小鼠興奮之影響………………………57
(六) 柳杉葉部精油物對小鼠鎮痛之影響(冰醋酸扭體試驗) ………………59
(七) 柳杉葉部精油物對小鼠熱刺激甩尾法之影響………………………60
(八) 柳杉葉部精油對戊四唑誘導驚厥小鼠之影響………………………62
四、柳杉葉部精油對小鼠之口服急毒性試驗結果 ………………………64
(一) 柳杉葉部精油對小鼠體重變化之影響………………………………64
(二) 柳杉葉部精油對小鼠血液學與血清肝及腎臟功能指數變化………68
(三) 柳杉葉部精油口服急毒性試驗對小鼠臟器之變化…………………72
伍、結論 ……………………………………………………………………79
陸、參考文獻 ………………………………………………………………80
柒、附錄 ………………………………………………………90
附錄1. LabDiet® 5001 飼料成分……………………………90
附錄2. 柳杉葉部精油口服急毒性試驗第14天對照組個別雄鼠之組織病理檢查………91
附錄3. 柳杉葉部精油口服急毒性試驗第14天對照組個別雌鼠之組織病理檢查……………………92
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