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-azinobis(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400μmol/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20μmol/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。
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-azinobis(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 400μmol/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 20μmol/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.
作者
张婧菲
韩红丽
沈明明
张莉莉
王恬
ZHANG Jingfei;HAN Hongli;SHEN Mingming;ZHANG Lili;WANG Tian(College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)
出处
《食品科学》
EI
CAS
CSCD
北大核心
2020年第13期96-105,共10页
Food Science
基金
国家自然科学基金青年科学基金项目(31601973)
江苏省科技计划项目-青年基金项目(BK20160739)
中央高校基本科研业务费专项资金项目(KJQN201707)。
