大蒜水溶性成份之抗氧化活性研究

Studies On The Antioxidant Activity of Garlic (Allium sativum) Water-soluble Constituents

狄亞莉(Ardhia Deasy Rosita Dewi)

指導教授 ： Wen-Ling Shih ； Dr.Ir. Joni Kusnadi,M.Si

國立屏東科技大學/農學院/生物科技研究所/碩士(2016年)

摘要

細胞內含有大量自由基造成正常細胞代謝混亂，最後導致各種疾病。大蒜是一種富含有機硫的天然物，被視為最常見的醫療試劑，其含有抗氧化活性可對抗疾病，而有機硫能有益身體健康的效果，特別是抑制活性氧。有趣的是，大蒜成分中的N-乙酰半胱氨酸（NAC），為大蒜水溶性成分之一，在DPPH自由基清除試驗中，具有高度抗氧化活性。本研究製備大蒜的水溶性成分和大蒜油並進行抗氧化活性之比較。使用LC-MS鑑定大蒜水溶性成分，獲得5種成分，即N-乙酰半胱氨酸，半胱氨酸丙氨酸，苯酚-2-2苯並噁唑基和兩種未知成分。接著用GC-MS分析大蒜油組成，獲得有機硫化物二烯丙基二硫（DADS）佔總成分的53.81％、二烯丙基三硫化物（DATS）佔31.49％，以及D-檸檬烯占14.71％。使用DPPH測試抗氧化活性，結果顯示:大蒜水溶性成分之自由基清除率70 ± 0.02% 較大蒜油58 ± 0.07% 具有高度抗氧化活性。巨噬細胞株RAW264.7細胞存活率試驗中，大蒜水溶性成分IC10 355µg/ml與大蒜油IC10 5.37µg/ml相比，對細胞較無毒性。使用過氧化氫誘導RAW264.7細胞產生活性氧測試，結果顯示，大蒜水溶性成分與大蒜油相比能有效降低活性氧產生大蒜水溶性和大蒜油可藉由激活抗氧化酶HO-1來減少氧來減少氧化壓力。當大蒜油10 µg/ml和大蒜水溶性萃取物400 µg/ml處理細胞4小時後，可透過Nrf2的表現來活化HO-1。

關鍵字

抗氧化活性, 大蒜水溶性, 大蒜油, 活性氧, HO

並列摘要

The abundance of free radical in cells leads to disrupted normal cellular metabolism eventually various diseases. Garlic is regarded as the most common medical agent containing organosulfur and phytochemical compounds against diseases. It is a natural source which has abundant organosulfur constituents. Organosulfur has beneficial health effects particularly, inhibiting Reactive Oxygen Species (ROS). Interestingly, among the constituents present in garlic, N-acetylcysteine (NAC) posseses the highest antioxidant activity when using DPPH radical scavenging analysis. Presence of NAC in garlic water-soluble may complete bioactive compounds in garlic water-soluble counteracting ROS. Garlic water-soluble constituents were identified by LC-MS and 5 constituents were found, namely N-acetylcysteine, cysteinyl-alanine, phenol-2-2-benzoxazolyl and 2 constituents unknown. The GC-MS chromatogram also showed 3 main constituents present in garlic oil as diallyl disulphide (DADS) 53.81%, diallyl trisulphide (DATS) 31.49% and D-limonene 14.71%. Subsequently, antioxidant activity determined by using DPPH assay showed that garlic water-soluble extract had higher DPPH scavenging activity 70 ± 0.02% compared with garlic oil 58 ± 0.07%. The survival of macrophage cell line RAW 264.7, indicated that garlic water-soluble IC10 355µg/ml was less toxic than garlic oil IC10 5.37µg/ml. Hydroperoxide was applied to induce Reactive Oxygen Species (ROS) in RAW 264.7 cells. ROS assay proved that ROS production cells were significantly reduced in garlic water-soluble extract compared with garlic oil FSU. Garlic water-soluble extract and garlic oil are important for reduce oxidative stress by activating antioxidant enzymes mainly HO-1. Maximum time for activation of HO-1 through induction of Nrf2 was achieved at 4 h when 10 µg/ml of garlic oil and 400 µg/ml of garlic water-soluble were induced.

並列關鍵字

Antioxidant activity, Garlic water-soluble, Garlic oil, Reactive Oxygen Species, HO-1

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