姜黄素类化合物的体外抗氧化活性及其对鸡红细胞氧化损伤的保护作用

doi:10.7506/spkx1002-6630-20190629-406

摘要/Abstract

摘要： 本实验旨在通过自由基清除实验和红细胞氧化应激模型比较3 种姜黄素（curcumin，CUR）的体外抗氧化活性。在自由基实验中，选取CUR、去甲氧基姜黄素（demethoxycurcumin，DUR）和双去甲氧基姜黄素（bisdemethoxycurcumin，BUR）为研究对象，测定其2,2’-联氮-双-3-乙基苯并噻唑啉-6-磺酸（2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)，ABTS）阳离子自由基、1,1-二苯基-2-三硝基苯肼（1,1-diphenyl-2-picrylhydrazyl，DPPH）自由基、羟自由基（·OH）和超氧阴离子自由基（O2－·）清除率，铁离子还原/抗氧化能力（ferric-reducing antioxidant power，FRAP）和脂质过氧化抑制率。同时，选取人工合成的抗氧化剂二丁基羟基甲苯（butylated hydroxytoluene，BHT）和丁羟茴醚（butylated hydroxyanisole，BHA）作为参照物。结果显示，3 种CUR均表现出良好的抗氧化活力，能有效清除ABTS阳离子自由基、DPPH自由基、·OH和O2－·，且其清除率呈现明显的浓度依赖性。与BHT相比，DUR对DPPH自由基的清除率显著提高（P＜0.05），CUR对DPPH自由基的清除率则显著降低（P＜0.05）。与BHA和BHT相比，3 种CUR对·OH的清除率均有显著降低（P＜0.05）。与CUR相比，DUR和BUR对·OH的清除率显著提高（P＜0.05）。与DUR相比，BUR和CUR对O2－·的清除率显著提高（P＜0.05）。在浓度为400 μmol/L时，BHA的FRAP最大，BUR的脂质过氧化抑制率最高。与CUR相比，BUR和DUR对脂质过氧化反应的抑制率显著提高（P＜0.05）。在红细胞氧化应激实验中，研究了3 种CUR对2,2’-偶氮二(2-甲基丙基咪)二盐酸盐（2,2’-azobis(2-amidinopropane) dihydrochloride，AAPH）处理后的红细胞溶血率、丙二醛（malondialdehyde，MDA）含量和超氧化物歧化酶（superoxide dismutase，SOD）活力的影响。结果显示，不同处理时间和不同浓度的3 种CUR均可明显抑制AAPH诱导的红细胞溶血作用，降低MDA含量和提高SOD活力；且这种保护作用呈现浓度依赖性和时间依赖性。孵育5 h后，在20 μmol/L时，与CUR组相比，DUR和BUR组的红细胞溶血率显著降低（P＜0.05）；与CUR和DUR组相比，BUR组的红细胞MDA含量显著降低（P＜0.05）；与BUR组相比，CUR和DUR组的红细胞SOD活力显著升高（P＜0.05）。结论：与CUR类似，BUR和DUR均表现出良好的抗氧化活性。3 种CUR能有效地清除自由基，具有较高的FRAP和脂质过氧化抑制率，可保护红细胞免受AAPH诱导的氧化损伤。就ABTS阳离子自由基、·OH和O2－·清除率和脂质过氧化抑制率而言，BUR的活性最佳。在缓解红细胞氧化损伤方面，DUR和BUR的抗氧化作用较CUR相比更显著。

关键词: 姜黄素, 去甲氧基姜黄素, 双去甲氧基姜黄素, 自由基, 鸡红细胞, 抗氧化

Abstract: The aim of the present study was to investigate the in vitro antioxidant activity of three curcuminoids in terms of radical scavenging capacity and protective effect against oxidative stress-induced damage in chicken erythrocytes. The scavenging capacity of curcumin (CUR), demethoxycurcumin (DUR) and bisdemethoxycurcumin (BUR) against 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) radical cation, 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical, hydroxyl radical (·OH), and superoxide anion O2-· radical were evaluated as well as their ferric-reducing antioxidant power (FRAP) and lipid peroxidation inhibitory potential. The synthetic antioxidants butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA) were used as reference standards. The results showed that all three curcuminoids exhibited strong ABTS, DPPH, ·OH and O2-· scavenging capacity in a dose-dependent manner. Compared with BHT, the DPPH radical scavenging capacity of DUR was significantly increased, while that of CUR was significantly decreased (P < 0.05). The ·OH scavenging capacity of the three curcuminoids was significantly decreased when compared with both BHA and BHT (P < 0.05). BUR and CUR had stronger ·OH scavenging activity than did CUR. Compared with DUR, the O2-· scavenging activity of BUR and CUR was significantly increased (P < 0.05). When used at a concentration of 400 μmol/L, BHA had the highest FRAP value and BUR exhibited the strongest inhibition of lipid peroxidation. Compared with CUR, BUR and DUR were significantly more effective against lipid peroxidation (P < 0.05). Moreover, all the curcuminoids significantly suppressed 2,2-azobis(2-amidinopropane) dihydrochloride (AAPH)-induced hemolysis of chicken erythrocytes, as well as decreased MDA level and increased superoxide dismutase (SOD) activity in a dose- and time-dependent manner. After 5 h incubation, DUR and BUR at 20 μmol/L showed lower hemolysis rate when compared with CUR. Compared with those treated with CUR and DUR, the MDA level and SOD activity in BUR-treated cells were significantly decreased (P < 0.05). In conclusion, DUR and BUR, similar to CUR, have good antioxidant activity. The three curcuminoids can sufficiently scavenge free radicals, have high FRAP values and potent inhibitory activity against lipid peroxidation, and can protect erythrocytes from AAPH-induced oxidative stress damage. Among these, BUR exhibits the highest antioxidant activity in free radical (including ABTS, ·OH and O2-·) scavenging and lipid peroxidation assays. DUR and BUR are more effective in protecting erythrocytes from oxidative stress damage than CUR.

Key words: curcumin, demethoxycurcumin, bisdemethoxycurcumin, free radical, erythrocytes, antioxidant

中图分类号:

张婧菲, 韩红丽, 沈明明, 张莉莉, 王恬. 姜黄素类化合物的体外抗氧化活性及其对鸡红细胞氧化损伤的保护作用[J]. 食品科学, 2020, 41(13): 96-105.

ZHANG Jingfei, HAN Hongli, SHEN Mingming, ZHANG Lili, WANG Tian. In Vitro Antioxidant Activity of Curcuminoids and Their Protective Effects against Oxidative Stress-Induced Damage of Chicken Erythrocytes[J]. FOOD SCIENCE, 2020, 41(13): 96-105.

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姜黄素类化合物的体外抗氧化活性及其对鸡红细胞氧化损伤的保护作用

In Vitro Antioxidant Activity of Curcuminoids and Their Protective Effects against Oxidative Stress-Induced Damage of Chicken Erythrocytes

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