臺灣博碩士論文加值系統

以當歸補血湯的單味藥材當歸:黃耆 (1:5) 和鐵皮石斛等進行清除α, α-diphenyl-β-picrylhydrazyl(DPPH)自由基之體外活性評估。本實驗針對此合方所含的有效成分黃耆皂苷(astragaloside)、阿魏酸(ferulic acid)及石斛多醣(Dendrobium candidum Polysaccharides)進行濃度分析。
當歸補血湯水萃及酒精萃取物所含的有效成分阿魏酸(ferulic acid)進行分析，結果得知當歸補血湯之水萃及酒精萃取物所含阿魏酸之含量分別為7.25±5.47與84.51 ± 11.31 g/g，以當歸補血湯酒精萃取物含量最高。而石斛多醣吸光度測試，最合適吸光值的濃度為300 g /mL，以酚硫酸法所得的葡萄糖標準曲線值換算，300 g 粗萃石斛多醣等同含glucose 85 g。當歸補血湯與鐵皮石斛具有清除DPPH自由基之抗氧化能力但清除效力均較正對照組維生素C較弱。
本研究在細胞毒性分析 (MTT assay)中發現，當歸補血湯酒萃物與石斛多醣合方，可明顯增加細胞之存活率，且對於大鼠眼部眼壓及視網膜病變具有明顯之保護作用。由高血糖大鼠血糖調降的試驗中，可明顯知道石斛多醣加當歸補血湯合方具有更佳之血糖調降效果。最後藉由生化檢查數據及病理切片觀察所得結果，服用當歸補血湯與鐵皮石斛合方對大鼠之腎臟及肝臟具有相當的保護能力，故可當作於日後改善人體預防高血糖及視網膜病變服用之參考。

The research used single dose including Danggui Buxue Tang and Angelica: Astragalus (1:5), and Dendrobium candidum to assess the efficiency in vitro. The analysis is the reduction of α-diphenyl-β-picrylhydrazyl (DPPH) radical of the Dendrobium candidum Polysaccharides. Also, the concentration analysis have made, regarding to the active ingredients of Astragaloside contained in this prescription, ferulic acid and Dendrobium candidum Polysaccharides.
We analyzed the active ingredients of ferulic acid ,which is the extract from water and alcohol in Danggui Buxue Tang. The results show that the content of ferulic acids were 7.25 ± 5.47 from water extract and 84.51 ± 11.31 g/g from alcohol extract, respectively. The content of ferulic acids from alcohol extract is higher. In the absorbance test of the Dendrobium candidum Polysaccharides, we had found that the most suitable concentration of the light absorption value is 300 g / mL in the OD (optical density) . To transfer the concentration of the light absorption value 300 g /mL of crude Dendrobium candidum Polysaccharides, according to a glucose standard curve values obtained by the phenol-sulfuric acid method, is equivalent to glucose 85 g/mL. Danggui Buxue Tang and Dendrobium candidum has show the capacity of reduction DPPH radical but the clear effect is weaker than the control group which given vitamin C.
In the cytotoxicity assay (MTT assay), the research found that the combination of Danggui Buxue Tang extracts by alcohol and Dendrobium candidum Polysaccharides can significantly increase the survival rate of cells, and show protective effects in the rat intraocular pressure and retinal eye disease. The combination obviously has the better efficacy of blood sugar lowering test in hyperglycemic rats. Finally, we checked the data by biochemical and pathological observations results to find the considerable protective effect of kidney and liver in rats with the combination medicine treatment. These findings show reference to improve, even prevent, the hyperglycemia and retinopathy for clinical treatment.

目 錄

中文摘要 I
Abstract II
誌 謝 IV
目 錄 V
表 目 錄 VII
圖 目 錄 VIII
第一章 前言 1
第二章 總論 6
第一節 當歸補血湯文獻回顧 6
第二節 當歸文獻回顧 7
第三節 黃耆文獻回顧 10
第四節 鐵皮石斛文獻回顧 14
第五節 糖尿病之簡介 16
第六節 糖尿病視網膜病變之簡介 20
第七節 研究目的 24
第三章 材料與方法 25
第一節 實驗動物 25
第二節 實驗儀器 25
第三節 實驗試劑與藥品 26
第四節 實驗方法 28
一、 概述 28
二、 當歸補血湯單味藥材，當歸與黃耆之基源鑑定 30
三、 當歸補血湯(當歸：黃耆＝1：5)之成分萃取 33
四、 鐵皮石斛之基源鑑定 33
五、 鐵皮石斛中粗萃石斛多醣之成分萃取 34
六、 當歸補血湯酒萃、水萃及粗萃石斛多醣之抗氧化試驗 34
七、 葡萄糖利用功效評估 35
八、 當歸補血湯與鐵皮石斛合方之細胞毒性分析(MTT assay) 36
九、 當歸補血湯與鐵皮石斛合方於糖尿病大鼠中，血糖調降作用之評估 39
十、 離體器官的病理切片觀察與試驗 41
十一、 統計方法 41
第四章 結果 42
第一節 當歸補血湯藥材(當歸與黃耆)及鐵皮石斛之基源鑑定 42
第二節 萃取產率 43
第三節 定性或定量分析 44
第四節 當歸補血湯酒萃、水萃及石斛多醣粗萃物之抗氧化試驗 45
第五節 葡萄糖利用評估 45
第六節 當歸補血湯與鐵皮石斛合方之細胞毒性分析 (MTT assay) 46
第七節 當歸補血湯與鐵皮石斛合方之改善血糖調降作用之功效評估 47
第五章 討論 51
第六章 結論 54
參考文獻 55




表 目 錄

表一 當歸補血湯水萃及乙醇萃取所得之粗萃物重量及產率 68
表二 鐵皮石斛中粗萃石斛多醣之萃取重量及產率 69
表三 當歸補血湯水萃及酒精萃取物經HPLC分析並對照其阿魏酸(ferulic acid)之檢量線，所得的每克藥材所含的單位含量 70
表四 Wistar大鼠每日平均攝食量及飲水量評估表 71
表五 連續分別給予石斛多醣(DC)、當歸補血湯酒萃物(DBT)和石斛多醣加當歸補血湯酒萃物(DBT-DC)八週對高果糖飲食大鼠血清生化含量變化 72
表六 連續分別給予石斛多醣(DC)、當歸補血湯酒萃物(DBT)和石斛多醣加當歸補血湯酒萃物(DBT-DC)八週對高果糖飲食大鼠血球計數值含量變化 73
表七 連續分別給予石斛多醣(DC)、當歸補血湯酒萃物(DBT)和石斛多醣加當歸補血湯酒萃物(DBT-DC)八週對高果糖飲食大鼠血液中葡萄糖、胰島素、胰島素抗性指數(HOMA-IR score)和葡萄糖-胰島素指數(ISI)含量變化 75










圖 目 錄

圖一 當歸藥材外觀圖 76
圖二 黃耆與鐵皮石斛藥材外觀圖 77
圖三 當歸及黃耆之生品薄層層析(TLC)圖 78
圖四 阿魏酸(ferulic acid) 檢量線圖、阿魏酸標準品圖譜和當歸補血湯酒萃圖譜及當歸補血湯水萃圖譜 79
圖五 酚-硫酸法所得的葡萄糖檢量線圖 80
圖六 當歸補血湯酒萃、水萃及石斛多醣粗萃物之清除DPPH自由基之抗氧化能力分析圖 81
圖七 葡萄糖利用評估 82
圖八 當歸補血湯與鐵皮石斛合方之細胞毒性分析 (MTT assay) 分析圖 83
圖九 大鼠眼壓分析圖 84
圖十 大鼠眼球視網膜照片 85
圖十一 Wistar大鼠每日平均體重變化分析圖 86
圖十二 口服葡萄糖耐量試驗 (oral glucose tolerance test; OGTT) 87
圖十三 大鼠肝臟病理切片圖 88
圖十四 大鼠腎臟病理切片圖 89

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