- 學位論文
台灣金線連之指標成分的定量及其調控果糖誘發高血糖症小鼠之血糖作用的探討
Quantification of the index components of Anoectochilus formosanus and study on its blood glucose regulation in fructose-induced hyperglycemic mice
摘要
中文摘要 隨著飲食習慣的改變，食物的精緻化，導致第二型糖尿病的罹患率上升，其主因多為胰島素阻抗而產生高血糖症，高血糖會增加體內活性氧化物含量，造成體內的氧化壓力上升，近年來已有報告指出氧化壓力容易導致胰島細胞受損，加重胰島素阻抗性。除了飲食以及生活方式的改變能改善胰島素阻抗，許多植物之天然成分已知具有生理活性也能有效地輔助疾病的控制。本研究目的是利用不同處理得到金線連甲醇萃取物，測定其指標成分以及抗氧化物質的含量，並進一步探討利用果糖誘發小鼠產生之高血糖症狀及探討金線連萃取物是否具有調控小鼠血糖之效果。 金線連經甲醇萃取後，再利用正己烷、乙酸乙酯、正丁醇以及水的不同極性溶媒來分劃，得到水層萃取物比例最高。再利用高壓液相層析來分析指標成分並定量每克粗萃取物含異鼠李素(isorhamnetin)及其醣苷(Isorhamnetin 3-O-β-D glucoside)以及金線連醣苷(kinsenoside)的量，分別為0.044±0.01、0.177±0.12及170.00±16.34 mg/g。金線連的特有醣類結構物質金線連醣苷，由於其水溶性，分佈在正丁醇的部分最多(265.7±24.40 mg/g)。在自由基清除作用方面，利用電子順磁共振法來偵測不同濃度(10 μg/ml~200 μg/ml)金線連乙酸乙酯層的結果，與濃度成正比，濃度上升清除自由基的能力也就上升，在200 μg/ml濃度下具有清除60%自由基的能力。 在動物實驗方面，我們將C57BL/6小鼠分為兩組，控制組為飲水，實驗組為飲用高果糖溶液，果糖誘發小鼠產生肥胖與高血糖症，實驗組小鼠在誘發第5週後空腹血糖值與控制組就有明顯統計上差異(p<0.05)，且在第8週實驗組小鼠體重與控制組有明顯統計上差異(p<0.05)。我們在誘發第22週進行葡萄糖耐受試驗，實驗組以及控制組，分別投予食鹽水、金線連萃取物10、25、50 mg/kg與Metformin 50mg/kg。在控制組小鼠投予金線連萃取物和空白組比較血糖濃度曲線下面積並沒有統計上的差異，可知金線連萃取物對正常小鼠的血糖值並沒有影響。在實驗組小鼠中，投予金線連萃取物隨著給藥劑量上升而血糖曲線下面積下降，其中25與50 mg/kg血糖曲線下面積比未投藥組來得小(p<0.001)，顯示金線連具有調降血糖之功能，且調降血糖的功能與劑量成正比。投予金線連萃取物50 mg/kg劑量與Metformin 50 mg/kg的曲線下面積沒有統計上差異，顯示兩者調降血糖功能相當。
並列摘要
英文摘要 As modern world lifestyle comes, people have different way of habits. Refined carbohydrate food and lack of exercise lead to a raise of metabolic disease- type 2 diabetes mellitus, which is characterized as the insulin resistance and leaded to hyperglycemia. In recent years, hyperglycemia was found to induce the generation of reactive oxygen species (ROS) and enhance oxidative stress. The oxidative stress-induced cell damage in pancreatic islet cell and aggravate insulin resistance. Modifications of diet and lifestyle are widely regarded as the cornerstones of treatment of insulin resistance. In addition, many nature occurring ingredients have shown their accompanied effects on disease control. The aims of this study is to quantitate the index components and evaluate free radical scavenging capacity of Anoectochilus formosanus extracts (AFE), to establish the hyperglycemia animal model by fructose and to investigate the blood glucose regulation of AFE on hyperglycemia. Anoectochilus formosanus Hayata was extracted with methanol and partition with organic solvents, n-hexane, ethyl acetate and n-butanol. High performance liquid chromatography (HPLC) was then used to determine the index components, isorhamnetin、Isorhamnetin 3-O-β-D glucoside and kinsenoside in freezing dry AFE, in which contained 0.04±0.01、0.18±0.12 and 170.00±16.34 mg per gram respectively. Kinsenoside, the peculiar glycoside of Anoectochilus formosanus is most distributed in n-butanol layer (265.7±24.40 mg). The free-radical scavenger capacity of AFE (10 μg/ml~200 μg/ml) was evaluated by electron paramagnetic resonance (ESR), and its clearance ability is along with the concentration of AFE. It has shown up to 60% of clearance of free radical in high (200 μg/ml) concentration of AFE. In animal model experiments, C57BL/6 mice were divided into two groups, control group and experimental group, which drink tap water and high fructose solution separately. Hyperglycemia and obesity mice model was successfully developed in experimental group Both groups were given separately with normal saline, AFE 10、25、50 mg/kg and metformin 50 mg/kg. In control group treated with AFE and normal saline, there has no significant statistical difference in incremental area under the blood glucose response curve (AUC) after intraperitoneal glucose tolerance test. It indicated that AFE did not affect the glucose level in control group. In experimental group, the AUCs were decrease in mice treated with AFE in a dose-dependent manner. A significant statistical difference exists in AUCs of mice treated with AFE 25 and 50 mg/kg compared to those with normal saline (p<0.001). AFE show its obviously blood glucose lowering effect in hyperglycermic mice. Comparison to metformin 50 mg/kg, there has no significant statistical difference in AUCs with AFE 50 mg/kg . The results suggested that AFE has blood glucose regulative effect and has a similar blood glucose lowering ability to diabetic drug-metformin with a dose of 50 mg/kg.
參考文獻
延伸閱讀
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