 |
| 系統識別號 |
U0005-1808200614361200 |
| 論文名稱(中文)
|
板藍萃取物在抗血癌、抑菌、抗假性狂犬病病毒及基因毒性之研究
|
| 論文名稱(英文)
|
Studies of Isatis indigotica on antileukemia, antitimicrobial, anti-PR virus and genotoxic effects |
| 校院名稱 |
中興大學 |
| 系所名稱(中) |
獸醫病理學研究所 |
| 系所名稱(英) |
Graduate Institute of Veterinary Pathology |
| 學年度 |
94 |
| 學期 |
2 |
| 出版年 |
95 |
| 研究生(中文) |
廖恬菱 |
| 研究生(英文) |
Tian- Ling Liao |
| 電子信箱 |
vminverse17@yahoo.com.tw |
| 學號 |
79347011 |
| 學位類別 |
碩士 |
| 語文別 |
中文 |
| 口試日期 |
2006-06-28 |
| 論文頁數 |
110頁 |
| 口試委員 |
委員-劉正義
委員-龐飛
委員- 康照洲
委員-陳世雄
指導教授-廖俊旺
|
| 中文關鍵字 |
板藍根
大青葉
人類血癌細胞
紙錠擴散法
基因毒性
安氏試驗
小鼠微核試驗
|
| 英文關鍵字 |
Isatis indigotica
HL-60
disc diffusion assay
genotoxicity
Ames test
micronucleus test
|
| 學科別分類 |
學科別＞醫學與生命科學＞動物學
|
| 中文摘要 |
板藍(Isatis indigotica Fortune)屬十字花科植物，其乾燥根及乾燥葉為常用之中藥板藍根、大青葉。傳統用於防治流行性感冒、傳染性肝炎、腮腺炎、慢性白血病及皰疹等。近年來有許多研究文獻在探討板藍的功效及其有效組成份的分析。已有研究顯示具有抗腫瘤，抑菌及致基因毒性作用的板藍成份分別為，靛玉紅(indirubin)、靛藍(indigo)及色胺酮(tryptanthrin)。為進一步了解板藍之藥理活性、抗病毒作用，及是否具基因毒性，本研究以本校農藝系自行栽種之板藍乾燥根甲醇(IEM)及水煎煮(IEW)萃取物；與大陸購得之板藍根甲醇(RM)、水(RD)及乙酸乙酯層(RE)萃取物，大青葉甲醇(LM)、水(LD)及乙酸乙酯層(LE)萃取物，另外與由大青葉所萃取出的成分靛玉紅(indirubin)作為比較。在抗血癌活性方面，靛玉紅(indirubin)、LE、RE及LM有明顯抑制人類血癌細胞(HL-60)生長，其半數抑制濃度(IC50)分別為：98 μg/mL、226 μg/mL、305 μg/mL及473 μg/mL。在抗菌實驗，使用豬隻常見病原菌Salmonella typhimurium, Salmonella choleraesuis, Escherichia coli, Actinobacillus pleuropneumoniae四株標準菌株，以紙錠擴散法(disc diffusion method)測試，皆無明顯抑菌作用。在抗病毒試驗中，使用豬假性狂犬病病毒(pseudorabies virus)感染豬腎細胞(PK-15)，可見大青葉萃取物抗病毒能力較板藍根明顯，尤以乙酸乙酯層更加顯著，顯示板藍萃取物具有毒殺PR病毒作用。基因毒性評估，以微生物基因突變Ames分析，結果顯示僅IEW對於TA98具有致突變性的可能，但在小鼠血球微核測試分析結果皆為陰性。綜合以上結果顯示，大青葉萃取物相較於板藍根萃取物抗血癌及抗PR病毒能力較具顯著作用，且以乙酸乙脂層之作用最明顯。在安全性方面，僅板藍根水煎煮萃取物(IEW)會引起沙門氏菌TA98菌種的基因突變，為一框構位移(frameshift)的突變作用。在血球微核試驗結果方面，顯示板藍對小鼠不會造成紅血球染色體異常。
|
| 英文摘要 |
The traditional Chinese medicine of Isatis indigotica (Cruciferae) is known as Ban-Lan-Gen and Da-Ching-Yeh, and are used for prevention and treatment of influenza, epidemic hepatitis, chronic leukemia and mumps, etc. Recently, numerous studies investigated the efficacy and its active compounds of Isatis indigotica. Indirubin, indigo and tryptanthrin are the compounds which possess antitumor, antimicrobial and genotoxic effects. In this study, we further evaluated the antileukemia, antimicrobial, anti-PR virus and genotoxic effects of methanol (IEM) and water (IEW)-extracted from root of Isatis indigotica that planted by Department of agronomy; dried root (R) and leaf (L) bought from China that extracted by using methanol (M), ethyl acetate (E) and distilled water (D). Results showed that the pure compound of indirubin displayed showed the most significant cytotoxicity on human leukemia cell line (HL-60) among these extracts, the IC50 was 98 μg/ml, followed by LE (IC50= 226 μg/ml), RE (IC50=305 μg/ml), LM (IC50= 473 μg/ml), respectively. These extracts did not show remarkable antimicrobial effects on Salmonella typhimurium, Salmonella choleraesuis, Escherichia coli, Actinobacillus pleuropneumoniae by using the disc diffusion method. Interestingly, numerous extracts of Da-Ching-Yeh had remarkable inhibition on pseudorabies (PR) virus replication in PK-15 cells, particularly extracts from ethyl acetate. The anti-PR virus effect of these extracts was proposed as virucidal activity. On the other way, IEW posed potential mutagenicity at the dose of 5 mg/plate on Ames test toward TA98 with or without liver S9 metabolism; however, no chromosome aberration was noted on micronucleus formation at dose of 0.2, 1 and 5 mg/kg bw of all extracts in ICR mice. From these results, extracts of Isatis indigotica had potential effects on anti-HL60 cancer cells, and anti-PR viral replication in PK-15 cells, extracts from Da-Ching-Yeh had better virucidal activity than Ban-Lan-Gen. Furthermore, crudes of ethyl acetate- extracts exhibited the highest efficacy among these extracts. Although IEW induced a frameshift mutation in TA98, all extracts of Isatis indigotica didn’t cause chromosomal aberration in mice.
|
| 論文目次 |
目 錄
頁次
中文摘要 ----------------------------------------------------------------------------- Ⅰ
英文摘要 ----------------------------------------------------------------------------- Ⅱ
目錄 ----------------------------------------------------------------------------- Ⅲ
圖次 ----------------------------------------------------------------------------- Ⅳ
表次 ----------------------------------------------------------------------------- Ⅴ
第一章 前言 -------------------------------------------------------------------------- 1
第二章 文獻探討 -------------------------------------------------------------------- 3
第一節 板藍之文獻資料 ----------------------------------------------------- 3
第二節 抗癌、抑菌及抗病毒活性之研究 -------------------------------- 5
第三節 抗突變性及基因毒性 ----------------------------------------------- 17
第三章 材料與方法 ----------------------------------------------------------------- 25
第一節 實驗材料與儀器 ----------------------------------------------------- 25
第二節 實驗設計 -------------------------------------------------------------- 28
第三節 試驗藥材 -------------------------------------------------------------- 28
第四節 抗血癌之細胞毒性試驗 ---------------------------------------------- 30
第五節 抑菌試驗 -------------------------------------------------------------- 32
第六節 抗病毒試驗 ----------------------------------------------------------- 33
第七節 抗致突變性試驗 ----------------------------------------------------- 37
第八節 基因毒性試驗 -------------------------------------------------------- 40
第九節 統計方法 ----------------------------------------------------------------- 41
第四章 結果 -------------------------------------------------------------------------- 43
第一節 板藍萃取物體外抗血癌、抑菌及抗病毒作用 -------------------- 43
第二節 板藍萃取物抗致突變性作用 -------------------------------------- 50
第三節 板藍萃取物之基因毒性 -------------------------------------------- 51
第五章 討論 -------------------------------------------------------------------------- 55
第六章 參考文獻 ----------------------------------------------------------------------- 95
|
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