臺灣博碩士論文加值系統

板藍根、大青葉及青黛為常用的中藥之一，板藍根之基原為為十字花科(Cruciferae)菘藍Isatis indigotica FORTUNE的乾燥根，功能為清熱解毒，涼血利咽；用於溫毒發斑，舌絳紫暗，痄腮，喉痺，爛喉丹痧，大頭瘟疫，丹毒，癰腫。大青葉為十字花科菘藍(Isatis indigotica FORTUNE)的乾燥葉，功能為清熱解毒，涼血消斑；用於溫邪入營，高熱神昏，發斑發疹，黃疸，熱痢，痄腮，喉痺，丹毒，癰腫。青黛之基原為爵床科(Acanthaceae)植物馬藍(Strobilanthes cusia (NEES) KUNTZE)、蓼科(Polygonaceae)植物蓼藍(Polygonum tinctorium AITON)、十字花科(Cruciferae)植物菘藍(Isatis indigotica FORTUNE)或豆科(Leguminosae)植物野木藍(Indigofera suffruticosa MILLER)的葉或莖葉經加工製得的乾燥粉末或團塊。其功能主治為清熱解毒，涼血，定驚；用於溫毒發斑，血熱吐衄，胸痛咳血，口瘡，痄腮，喉痺，小兒驚癇；而其主要成分為靛藍(indigo)及靛玉紅(indirubin)。根據調查，目前臺灣所用的板藍根、大青葉及青黛藥材來源皆不止一種，故本研究分別進行此三類藥材之品種鑑別、性狀鑑別、顯微鑑別、化學鑑別及藥理活性評估。
以生藥組織鑑別結果顯示，本研究收集台灣市售板藍根藥材廿五件中，馬藍(Strobilanthes cusia (NEES) KUNTZE)莖佔廿三件，馬藍(Strobilanthes cusia (NEES) KUNTZE)根及根莖與野木藍(Indigofera suffruticosa MILLER)莖各佔一件。市售大青葉藥材廿五件中，馬藍(Strobilanthes cusia (NEES) KUNTZE)之乾燥葉佔廿四件，而大青(Clerodendrum cyrtophyllum TURCZANINOW)之乾燥葉一件。至於大陸藥典收載之板藍根（菘藍根Isatis indigotica FORTUNE）大青葉（菘藍葉Isatis indigotica FORTUNE）及蓼大青葉(Polygonum tinctorium AITON)藥材，在台灣則未發現。
台灣市售30件青黛檢品，經檢驗後發現大部份為不含靛藍及靛玉紅之藍色粉末，不具草腥味，且經ICP-MS檢測其所含之Fe較正品青黛高。而臺灣中藥廠所用之青黛及當歸龍薈丸製劑則可檢驗出靛藍及靛玉紅。在HPLC定量分析方面，本研究以RP-18管柱，在UV 292 nm檢測波長下，利用甲醇與水當移動相，以線性梯度條件進行分析，得到迅速、靈敏、準確且再現性不錯之分離效果。而indigo及indirubin之最低檢出量為12 ng/mL。
急性毒性實驗結果顯示，腹腔注射給予高達1000 mg/kg劑量之靛藍及靛玉紅，未見動物死亡；腹腔注射給予菘藍根(II-R) 5000 mg/kg，亦未見動物死亡；而馬藍莖(SC-S)、馬藍根及根莖(SC-R)、馬藍葉(SC-L)及菘藍葉(II-L)之LD50依序分別為3134.60, 5076.66, 2742.00及2510.39 mg/kg。
由青黛萃取之主要成分靛藍及靛玉紅之藥理活性試驗結果顯示，在鎮痛方面，對醋酸誘發鼷鼠扭體反應，靛藍及靛玉紅均呈現劑量依存性的抑制作用；對福馬林誘發舔足的前期反應時間，靛藍及靛玉紅呈現劑量依存性的抑制作用，對福馬林誘發舔足的後期反應時間，僅靛玉紅投予50 mg/kg時，有明顯抑制作用。在抗炎方面，靛藍及靛玉紅對λ-carrageenin注入大鼠足蹠誘發浮腫有抑制作用。在抗腫瘤活性方面，靛藍及靛玉紅對人類喉癌細胞(KB-16)、老鼠血癌細胞(P-388)、人類肺癌細胞(A-549)及人類腸癌細胞(HT-29)未見生長抑制作用。
由以上初步結果顯示，靛藍及靛玉紅對數種腫癌細胞之生長無抑制作用，但皆具有鎮痛抗炎之作用；靛玉紅之鎮痛作用可能經由周邊神經系統或抑制疼痛物質釋放作用外，亦對中樞神經系統有影響。
在板藍根及大青葉藥材方面，馬藍莖(SC-S)、馬藍根及根莖(SC-R)、菘藍根(II-R)、馬藍葉(SC-L)及菘藍葉(II-L)對於醋酸誘發鼷鼠扭體反應皆有劑量依存性之抑制作用；在福馬林舔足鎮痛試驗方面，市售板藍根藥材馬藍莖(SC-S)、馬藍根及根莖(SC-R)、菘藍根(II-R)、馬藍葉(SC-L)及菘藍葉(II-L)，對於福馬林誘發舔足的前期及後期反應時間，均呈現劑量依存性的抑制作用，由此可知，此三種板藍根及二種大青葉藥材之鎮痛作用除對經由周邊之作用外，亦對中樞有影響。在抗炎方面，馬藍莖(SC-S)、馬藍根及根莖(SC-R)、菘藍根(II-R)、馬藍葉(SC-L)及菘藍葉(II-L)對l-carrageenan誘發大白鼠足蹠浮腫皆有抑制作用。
室溫下對正常大鼠肛溫變化之影響方面，馬藍莖(SC-S)、馬藍根及根莖(SC-R)、菘藍根(II-R)、馬藍葉(SC-L)及菘藍葉(II-L)，皆對大鼠產生降溫作用，且在90分鐘時降至最低，其後則慢慢回復，且具有劑量依存性之關係。對LPS誘發發燒大鼠之影響方面，在給藥物90分鐘後溫度變化最大，在46.88 mg/kg劑量時，馬藍莖(SC-S)、馬藍根及根莖(SC-R)及菘藍根(II-R)皆有統計上之意義；馬藍葉(SC-L)及菘藍葉(II-L)亦皆有劑量依存性抑制發燒現象。
在細胞毒性方面，馬藍莖(SC-S)、馬藍根及根莖(SC-R)、菘藍根(II-R)、馬藍葉(SC-L)及菘藍葉(II-L)對人類喉癌細胞(KB-16)、老鼠血癌細胞(P-388)、人類肺癌細胞(A-549)及人類腸癌細胞(HT-29)之生長皆未見抑制作用。
在藥物誘發肝損傷方面，馬藍莖(SC-S)、馬藍根及根莖(SC-R)、菘藍根(II-R)、馬藍葉(SC-L)及菘藍葉(II-L)對CCl4, APAP及D-GalN所導致sGOT及sGPT值急速上升皆有抑制作用，由病理切片圖亦可看出對藥物誘發之肝損傷亦有修復改善作用；但對於ANIT所誘發之肝損傷，三種板藍根藥材及二種大青葉藥材皆無明顯治療作用，但三種板藍根藥材及馬藍葉(SC-L)卻有利膽作用。
綜合本研究結果，臺灣所用板藍根及大青葉藥材皆為臺灣本產植物，雖然臺灣市售所用之板藍根大部份為馬藍莖(SC-S)，但其在鎮痛、抗炎、解熱及由不同藥物誘發肝損傷之治療作用方面皆具有一定之效果；至於大青葉藥材，雖然大陸藥典收載為菘藍葉(II-L)，但臺灣主要使用之馬藍葉(SC-L)在鎮痛、抗炎、解熱及治療肝損傷等藥理試驗亦顯示有良好之作用。馬藍(Strobilanthes cusia (NEES) KUNTZE)為臺灣本產之藥用植物，而大陸藥典收載之板藍根及大青葉臺灣均未產，以本研究結果，應可將馬藍開發成為一新的藥物。至於青黛藥材真偽之辨識，可以其是否具有草腥味簡單加以區分，而臺灣使用之偽品青黛其成分究竟為何？則值得進一步加以評估。

Ban-Lan-Gen, Da-Ching-Yeh and Ching-Dai are the commonly used Chinese herbs. The root of Isatis indigotica FORTUNE (Cruciferae), popularly known as Ban-Lan-Gen is used in traditional Chinese medicine for seasonal febrile diseases, pestilence, mumps, eruptive diseases, inflammatory diseases with redness of skin, sorethroat, etc.. The leaf of Isatis indigotica FORTUNE (Cruciferae), popularly known as Da-Ching-Yeh is used in traditional Chinese medicine to clear away heat and toxic material, cool blood and eliminate eruption and for heat-syndrome with high fever, thirst, restlessness and skin eruptions, especially for pestilence. Now it is usually used for influenza, epidemic cerebrospinal meningitis, encephalitis B, viral pneumonia and mumps. It is also used for sorethroat, aphthae, inflammatory diseases with redness of skin, etc.. Ching-Dai is the dried powder prepared from the leaves and stems of Strobilanthes cusia (NEES) KUNTZE (Acanthaceae), Isatis indigotica FORTUNE (Cruciferae), Polygonum tinctorium AITON (Polygonaceaae) or Indigofera suffruticosa MILLER (Leguminosae). It is used to clear away heat and toxic material, cool the blood and disperse heat in the liver and used for febrile disease, hematemesis, hemoptysis, and mumps. Indigo and indirubin are the active ingredients of Ching-Dai.
According to our previous investigation, we found that the origins and commercial species of Ban-Lan-Gen, Da-Ching-Yeh and Ching-Dai were complicated in Taiwan. Thus, we used comparative pharmacognosy methods to compare the origin, morphology, microscopic characteristics, chemical constituents and pharmacological effects of commercially available species.
Twenty-five Ban-Lan-Gen specimens from crude drug stores in Taiwan were collected. After morphological identification, twenty-three were identified as the stems of Strobilanthes cusia, one as the root and rhizome of Strobilanthes cusia, and one as the stem of Indigofera suffruticosa. The root of Isatis indigotica, the species recorded in the pharmacopeia of PRC, was not seen in this study.
Twenty-five Da-Ching-Yeh specimens from crude drug stores in Taiwan were also collected. After morphological identification, twenty-four were identified as the leaves of Strobilanthes cusia, one as the leaf of Clerodendrum cyrtophyllum. The leaves of both Isatis indigotica and Polygonum tinctorium were not seen in this study.
Of the thirty samples collected from Taiwan area, most of Ching-Dai were found to be dyes without herbal smelly odor and no indigo and indirubin were found at all. The compound preparations, Dan-Kuei-Lung-Huei-Wan made by GMP pharmaceutical company in Taiwan were found to contain indigo and indirubin. The result of ICP-MS showed that the counterfeit Ching-Dai contained more Fe than the genuine Ching-Dai. In HPLC analysis, RP-18 column was used with methanol and water as mobile phase under linear gradient conditions and the UV detection wavelength was 292 nm. We got rapid, sensitive, accurate and reproducible results. The limits of detection of indigo and indirubin were 12ng/mL.
This study also evaluated the antinociceptive and antiinflammatory effects of indigo and indirubin extracted from Ching-Dai. The results showed that indigo and indirubin inhibited the writhing response induced by acetic acid with a dose-response. In addition, indigo and indirubin repressed the licking time on the early phase in mice, but only indirubin at the dose of 50 mg/kg inhibited the licking time on the late phase in the formalin test. Indigo and indirubin inhibited the inflammatory edema induced by l-carrageenin in the initial phase. However, neither indigo nor indirubin showed cytotoxicity against the KB-16, P-388, A-549 and HT-29 cell lines.
The above results indicated that indigo and indirubin had no influence on cancer cells, but had both antinociceptive and antiinflammatory effects. The antinociceptive effect of indirubin might due to the effects on both the peripheral and central nerve systems.
In this study, we also evaluated the antinociceptive, anti-inflammatory, antipyretic effects and cytotoxicity activity of Ban-Lan-Gen specimens (the stem of Strobilanthes cusia (SC-S), root and rhizome of Strobilanthes cusia (SC-R) and root of Isatis indigotica (II-R))and Da-Ching-Yeh specimens (the leaf of Strobilanthes cusia (SC-L) and leaf of Isatis indigotica (II-L)). The results showed that all the methanolic extracts of Ban-Lan-Gen specimens and Da-Ching-Yeh specimens significantly inhibited the writhing responses of mice and decreased the licking time on both the early and late phases of formalin test in a dose dependent manner. The antinociceptive effects of SC-S, SC-R, II-R, SC-L and II-L extracts not only influenced the peripheral system but also the central system. They also reduced the paw edema induced by l-carrageenan in rats. In addition, they potently attenuated pyrexia induced by lipopolysaccharide and the change of colonic temperature dropped to bottom at 1.5 h with each dose. They also produced a dose-related fall in colonic temperature at room temperature in normal rats. However, all three materials did not show cytotoxicity against the KB-16, P-388, A-549 and HT-29 cell lines.
In this study, SC-S, SC-R, II-R, SC-L and II-L extracts showed different therapeutic effects against liver damage induced by carbon tetrachloride, acetaminophen, a-naphthylisothiocyanate and D-galactosamine. All extracts inhibited the high values of sGOT and sGPT induced by different drugs and the photomicrographs of liver section taken from rats showed their improvement of liver damage. Because of the differences in their mechanisms of injury, it was indicated that they exerted their therapeutic effects through more than one mechanism. However, SC-S, SC-R, II-R, SC-L and II-L extracts did not have therapeutic effects on ANIT-induced acute liver damage in rats, but SC-S, SC-R, II-R and SC-L could promote biliary elimination.
To sum up, the crude drugs of Ban-Lan-Gen and Da-Ching-Yeh used in Taiwan were native plants. Although the major Ban-Lan-Gen specimens in Taiwan were the stems of Strobilanthes cusia, it had the antinociceptive, anti-inflammatory, antipyretic effects and therapeutic activities against liver damage induced by different drugs. As to Da-Ching-Yeh species, the leaves of Strobilanthes cusia were commonly used in Taiwan. They also showed the antinociceptive, anti-inflammatory, antipyretic effects and therapeutic activities against liver damage induced by different drugs.
Isatis indigotica was not produced in Taiwan. According to the above results, the activities of the stem, root, rhizome and leaves of Strobilanthes cusia were similar to the root and leaves of Isatis indigotica. Therefore, it was worthwhile to develop Strobilanthes cusia as a new medicine. Genuine Ching-Dai could be easily distinguished by its herbal smelly odor. The exact constituents of the two counterfeit Ching-Dai is worthwhile for further investigation.

封面
謝辭
目錄
圖目錄
表目錄
略字表
中文摘要
英文摘要
第一章　前言
第二節　總論
第一節　本草學考察
第二節　藥用物學考察
第三節　化學成分整理
第四節　藥理作用文獻整理
第五節　藍染植物與加工技術考察
第三章　材料與方法
第一節　實驗試藥與儀器
第二節　實驗方法
第四章　結果
第一節　生藥組織鑑別
第二節　青黛製作與分析
第三節　藥理活性試驗
第五章　討論
第六章　結論
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