臺灣博碩士論文加值系統

金錢薄荷 (Glechoma hederacea) 為唇形花科植物，又名連錢草、活血丹、落地金錢等，為一傳統入藥與食用植物，民間傳用具有解熱利尿、活血化瘀等功效。先前以體外試驗探討金錢薄荷熱水萃取物 (HWG) 具清除自由基及抑制脂質過氧化之效力，並且證實不具有基因毒性，故本實驗接續以14天單一劑量毒性試驗和28天亞急性毒性試驗之動物餵食模式評估HWG之急性及亞急性毒性。
單一劑量毒性試驗，將SD大鼠隨機分為控制組、HWG低 (1 g / kg BW) 與高劑量組 (5 g / kg BW)，每組20隻 (雌、雄性各十隻)，實驗為期14天，期間觀察各組臨床症狀並紀錄體重和攝食量，犧牲後秤其臟器重量、分析血液生化值和血液常規。結果顯示，無論是觀察體重、攝食量、臨床症狀及臟器相對重量皆與控制組無統計學差異；在檢驗血液生化值和血液常規，於各組間均無毒性影響，故證實HWG對實驗動物不具急毒性反應，並可估計HWG之半數致死劑量 (LD50) 為5 g / kg BW以上。
在28天亞急性毒性試驗中，分為控制組、HWG低 (0.25 g / kg BW)、中 (0.5 g / kg BW) 與高劑量組 (1 g / kg BW)。結果顯示，觀察體重、攝食量和臨床變化皆無顯著差異，在臟器相對重量發現低與高劑量之雌鼠於脾臟顯著高於控制組和中劑量組，但觀察其外觀無異常腫大且重量仍在正常值範圍內；對於評估血液生化值和血液常規方面，在HWG劑量組與控制組間均無毒性影響，故HWG之添加對生物體內不會造成傷害，其無毒性顯示劑量 (no-observed-adverse-effect level, NOAEL) 為1 g / kg BW以上。
進一步分析雄鼠心、肝和腦部組織中抗氧化物質與酵素，包含維生素E (Vit E)、維生素C (Vit C)、麩胱甘肽 (GSH)、麩胱甘肽過氧化酶 (GPx)、觸酶 (CAT) 及超氧歧化酶 (SOD)，此外測定脂質過氧化物-丙二醛 (MDA) 含量及血漿中總抗氧化力 (TAS)。結果顯示，HWG實驗組有顯著提升腦中Vit E、心臟中Vit C含量及SOD之活性，並可維持動物臟器組織中GSH之含量及GPx和CAT之活性，且不會造成MDA之生成增加及TAS之降低。綜合上述，HWG對生物體內不具急性及亞急性毒性，且亦可藉由提升體內抗氧化保護機制來降低氧化傷害，未來將進一步對HWG做更多之生理活性探討。

Glechoma hederacea belongs to the Lamiaceae family, was used for treating diuresis and stimulating blood circulation according to ancient Chinese prescription. Previously studies have demonstrated that hot water extract of Glechoma hederacea (HWG) possessed antioxidant and anti-mutagenic characteristics. However, the safety of HWG remains to be evaluated. The objective of this study was to evaluate acute oral toxicity and 28-days repeated feeding toxicity tests for HWG by using the oral gavage method in Sprague-Dawley (SD) rats.
In the acute toxicity test, a single dose of 1and 5 g / kg body weight (b.w.) HWG was given to SD rats, then the rats were observed for 14 days. No acute lethal effect at a maximal dose of 5 g / kg b.w. HWG was observed in rats. In the 28-days repeated feeding toxicity tests, the rats were administered daily by gavage at dose of 0.25, 0.5 and 1 g / kg b.w. HWG for 28 days. The results indicated that no significant toxic effect was found in the parameters of body weight, feed consumption, relative organ weights, as well as hematological and serum biochemical parameters between control group and the HWG-treated rats. The no-observed-adverse-effect-level (NOAEL) of HWG was more than 1 g / kg b.w. for subacute toxicity study.
Furthermore, in the 28-days repeated feeding toxicity tests was conducted to determine the effect of HWG on the oxidation–antioxidantion equilibrium of male rats. The results showed that HWG-treated significantly elevated the level of vitamin E in brain, vitamin C and the activity of superoxide dismutase in heart compared with the control group. The HWG-treated maintained the balance of the level of glutathione and the activities of catalase, glutathione peroxidase. Besides, the level of lipid peroxidation and plasma of total antioxidant status (TAS) showed that HWG-treated was not significantly changed compared with the control group. In conclusion, HWG not noly safe, but also can be elevated capable of antioxidant for rats. Further, the more physiological activity of HWG need to be investigated.

目錄 I
表目錄 IV
圖目錄 V
中文摘要 VI
英文摘要 VIII
第一章 前言 1
第二章 文獻回顧 3
第一節 金錢薄荷簡介 3
第二節 自由基與活性氧物質介紹 6
一、過氧化氫 (hydrogen peroxide, H2O2) 8
二、氫氧自由基 (hydroxyl radical, ‧OH) 8
三、超氧陰離子 (superoxide anion radical, ‧O2-) 8
四、單重態氧 (singlet oxygen, 1O2) 8
五、脂質過氧化物 (lipid peroxide, LPO) 8
第三節 生物體內抗氧化防禦系統 9
一、酵素性防禦系統 9
1. 超氧歧化酶 (superoxide dismutase, SOD) 9
2. 觸酶 (catalase, CAT) 10
3. 麩胱甘肽過氧化酶 (glutathione peroxidase, GPx) 10
二、非酵素性抗氧化防禦系統 10
1. 維生素E (Vit E) 10
2. 維生素C (ascorbic acid, Vit C) 10
3. 麩胱甘肽 (glutathione, GSH) 11
4. 類胡蘿蔔素 (β-carotene) 11
5. 泛醌 (ubiquinone) 11
第四節 食品安全性評估 12
一、食品安全性標準制定 12
二、食品安全性評估分類 12
(一) 基因毒性試驗分類 (Genotoxicity study) 13
(二) 餵食毒性試驗 (Feeding toxicity study) 14
第三章 研究目的 16
第四章 研究流程 17
第五章 材料與方法 18
第一節 實驗材料 18
第二節 實驗方法 21
一、金錢薄荷萃取物之製備 21
二、動物餵食實驗 21
1. 單一劑量口服毒性試驗 23
2. 28天餵食毒性試驗 23
3. 抗氧化物質與酵素活性測定 24
(1) 組織均質液之製備: 24
(2) 維生素C 含量測定 24
(3) 維生素E 含量測定 25
(4) glutathione測定 26
(5) 丙二醛 (MDA) 含量測定 27
(6) Superoxide dismutase (SOD) 活性測定 28
(7) Glutathione peroxidase (GPx) 活性測定 28
(8) catalase活性測定 29
(9) 總蛋白質含量之測定 29
(10) 血漿中總抗氧化力 (total antioxidant status, TAS) 之測定 30
第三節 統計方法 31
第六章 結果與討論 32
第一節 單一劑量口服毒性試驗 (急性毒性試驗) 32
一、管灌金錢薄荷水萃取物14天之體重與攝食量變化及其相對臟器重 32
二、管灌金錢薄荷水萃取物14天後之血液生化值 36
三、管灌金錢薄荷水萃取物14天後之血液常規檢查 38
第二節 28天連續餵食毒性試驗 (亞急性毒性試驗) 41
一、管灌金錢薄荷水萃取物28天之體重與攝食量變化及其相對臟器重 41
二、管灌金錢薄荷水萃取物28天後之血液生化值 45
三、管灌金錢薄荷水萃取物28天後之血液常規 46
四、管灌金錢薄荷水萃取物28天後之雄鼠主要器官中抗氧化物質及脂質過氧化物含量 50
五、管灌金錢薄荷水萃取物28天後之雄鼠主要器官中抗氧化酵素活性 53
六、管灌金錢薄荷水萃取物28天後之雄鼠血漿中總抗氧化力 55
第七章 結論 58
第八章 參考文獻 59

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