School of Chinese Pharmaceutical Sciences and Chinese Medicine Resources, College of Pharmacy, China Medical University, Taichung 404, Taiwan.
Food Chem. 2012 Dec 1;135(3):861-7. doi: 10.1016/j.foodchem.2012.05.082. Epub 2012 May 29.
This study was designed to investigate the antioxidant activities of sweet potato defensin (SPD1) in vitro and ex vivo. Antioxidant status [2,2'-azinobis[3-ethylbenzthiazoline-6-sulphonic acid (ABTS) assay], scavenging activity against DPPH (1,1-dipheny-2-picrylhydrazyl) radical method, reducing power method, Fe(2+)-chelating ability, FTC (ferric thiocyanate) method, and protection of calf thymus DNA against hydroxyl radical-induced damage were studied in vitro. The ex vivo experiments revealed that SPD1 could decrease the production of intracellular peroxide in HepG2 cells. Four peptides, namely GFR, GPCSR, CFCTKPC and MCESASSK for testing antioxidative activity, were synthesized according to tryptic hydrolysis simulation. In the TEAC assay CFCTKPC performed the best (13.5±0.3μmol TE/g dw), even better than reduced glutathione (7.3±0.2μmol TE/g dw). In the DPPH radical assay (%), [IC(50) (μM) (the concentration required for scavenging 50% activity)] CFCTKPC again had the highest antioxidant activity (IC(50) is 11.3±3.2μM) even better than reduced glutathione (IC(50) is 74.3±2.4μM). In the lipid peroxidation assay, once again CFCTKPC performed the best, with an IC(50) value of 0.5±0.0μM better than reduced glutathione (1.2±0.1μM). These findings mean that cysteine residue is most important in antioxidant activities. It was suggested that SPD1 might contribute its antioxidant activities against hydroxyl and peroxyl radicals.
本研究旨在体外和离体条件下研究甘薯防御素(SPD1)的抗氧化活性。在体外研究了抗氧化状态[2,2'-联氮双(3-乙基苯并噻唑啉-6-磺酸)(ABTS)测定法]、对 DPPH(1,1-二苯基-2-苦基肼)自由基的清除活性、还原能力法、Fe(2+)螯合能力、FTC(铁硫氰酸盐)法以及保护小牛胸腺 DNA 免受羟基自由基诱导的损伤。离体实验表明,SPD1 可降低 HepG2 细胞内过氧化物的产生。根据胰蛋白酶水解模拟,合成了四种肽,即 GFR、GPCSR、CFCTKPC 和 MCESASSK,用于测试抗氧化活性。在 TEAC 测定中,CFCTKPC 表现最佳(13.5±0.3μmol TE/g dw),甚至优于还原型谷胱甘肽(7.3±0.2μmol TE/g dw)。在 DPPH 自由基测定中(%),[IC50(μM)(清除 50%活性所需的浓度)]CFCTKPC 再次具有最高的抗氧化活性(IC50 为 11.3±3.2μM),甚至优于还原型谷胱甘肽(IC50 为 74.3±2.4μM)。在脂质过氧化测定中,CFCTKPC 再次表现最佳,IC50 值为 0.5±0.0μM,优于还原型谷胱甘肽(IC50 为 1.2±0.1μM)。这些发现意味着半胱氨酸残基在抗氧化活性中最重要。因此,SPD1 可能有助于其对羟基和过氧自由基的抗氧化活性。