以D型半乳糖誘導老化動物模式評估之天然物抗氧化活性研究

Study on Antioxidant Activities of Natural Products in D-Galatose-Induced Aging Mice

李泰霖(Tai-Lin Lee)

指導教授 ： 侯文琪

臺北醫學大學/藥學院/生藥學研究所/碩士(2012年)

https://doi.org/10.6831/TMU.2012.00234

摘要

薑黃素 (curcumin) 是植物薑黃 (turmeric) 內主要具活性的成分，包含抗發炎、抗氧化、神經保護等活性。本篇研究探討薑黃素及其二十三個結構衍生物改善D型半乳糖誘導之老化鼠學習記憶能力及其抗氧化活性。經由細胞存活率測試 (Resazurin assay) ，發現結構衍生物七號及八號在10 ?嵱下對於以6-羥基多巴胺誘導人類母細胞瘤SH-SY5Y之神經毒性有顯著的保護作用。在同樣的條件下，也發現結構衍生物七號及八號有抑制細胞內ROS產生的能力。
在抗氧化能力方面，ORAC (oxygen radical absorption capacity) 總抗氧化能力試驗中，發現結構衍生物七號及八號在20 ?嵱、40 ?嵱、80 ?嵱都有優於薑黃素的抗氧化能力。抑制AAPH誘導紅血球溶血試驗中，發現結構衍生物七號及八號在20 ?嵱、40 ?嵱、80 ?嵱對於抑制紅血球溶血的時間都較同濃度的薑黃素為長，顯示出較好的抗氧化活性。
在動物實驗上，以D型半乳糖 (1.2 g/Kg bw/day) 的劑量誘導老化。透過被動迴避試驗評估其改善記憶能力的效果，在本篇研究中結構衍生物七號及八號沒有明顯的增加記憶恐懼的時間，故在行為學上沒有明顯的表現。在血清及組織的ORAC總抗氧化能力的實驗中，餵食結構衍生物七號及八號的組別，相較於實驗對照組在血清裡的ORAC值有顯著的提升，但在全腦組織中則沒有效果。而在丙二醛含量試驗中，餵食結構衍生物七號及八號的組別，在血清裡的丙二醛含量，相較於實驗對照組，有顯著的減少；而在全腦組織中只有在高濃度組 (10 mg/Kg bw) 顯示出其減少脂質過氧化的能力。
而在抗發炎活性方面，利用LPS誘導RAW 264.7細胞株產生NO的方法，評估結構衍生物七號及八號抑制NO產生的能力。結構衍生物七號及八號在10 ?嵱下有顯著的減少NO產生，且皆較薑黃素為優。
在本篇研究中，發現薑黃素結構衍生物七號及八號皆具有抗氧化及抗發炎的活性，但是在改善記憶功能的行為學研究上則沒有效果。

關鍵字

d型半乳糖 ； 薑黃素 ； 老化 ； 氧化壓力

並列摘要

Curcumin is the primary bioactive constituent of turmeric, which is an herbal medicine used in Asia for a long time. It has been shown to possess anti-inflammatory, antioxidant, and neuroprotection. In this study, curcumin and 23 different kinds of curcumin analogs were evaluated for antioxidant, anti-inflammatory and the ability to improve the learning and memory function in D-galactose-induced aging mice. According to the results of cell viability assay, analog 7 and 8 showed stronger neuroprotection and ROS inhibition activities against 6-OHDA, neurotoxic agent, in SH-SY5Y cell lines than curcumin under concentration of 10 μM. Besides, analogs 7 and 8 also showed better oxygen radical absorption capacity (ORAC) and the inhibitory effect of RBC hemolysis induced by AAPH than curcumin under concentration of 20, 40, 80 ?嵱 and were selected as test candidates for animal model.
Taking 1.2g/Kg bw/day D-galatose subcutaneous injection on BALB/c male mice as the animal model for aging. By the step-through passive avoidance test, analog 7 and 8 didn’t improve the memory function at all. By ORAC and TBARS assay, analog 7 and 8 show the antioxidant activities through compare with the group D-galactose only on serum and whole brain. In anti-inflammatory activities, it was found that analog 7 and 8 could inhibit iNOS on whole brain and decrease NO production more than curcumin on RAW 264.7 induced inflammatory by LPS (1 ?慊/mL) .

並列關鍵字

d-galactose ； curcumin ； aging ； oxidative stress

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