Park Min-Jung, Han Ji-Sook
Department of Food Science and Nutrition, Pusan National University, Busan 609-735, Korea.
Prev Nutr Food Sci. 2013 Dec;18(4):227-33. doi: 10.3746/pnf.2013.18.4.227.
This study investigated the protective effect of the butanol (BuOH) fraction from fermented Laminaria japonica extract (BFLJ) on AAPH-induced oxidative stress in porcine kidney epithelial cells (LLC-PK1 cells). L. japonica was fermented by Aspergillus oryzae at 35±1°C for 72 h. Freeze-dried fermented L. japonica was extracted with distilled water, and the extracted solution was mixed with ethanol and then centrifuged. The supernatant was subjected to sequential fractionation with various solvents. The BuOH fraction was used in this study because it possessed the strongest antioxidant activity among the various solvent fractions. The BuOH fraction of fermented L. japonica had a protective effect against the AAPH-induced LLC-PK1 cells damage and increased cell viability while reducing lipid peroxidation formation and increased activities of antioxidant enzymes such as superoxide dismutase and glutathione peroxidase. The inhibitory effect of BFLJ on lipid peroxidation formation had a higher value of 0.11±0.01 nmol MDA at 100 μg/mL concentration in comparison with intact BuOH fraction showing 0.22±0.08 nmol MDA at the same concentration. Furthermore, BFLJ treatment increased glutathione concentration. GSH concentration in the cell treated with BFLJ of 100 μg/mL was 1.80 pmol/L×10(5) cells. These results indicate that BFLJ protects the LLC-PK1 cells against AAPH-induced cell damage by inhibiting lipid peroxidation formation and increasing antioxidant enzyme activities and glutathione concentration.
本研究考察了发酵海带提取物的丁醇(BuOH)组分(BFLJ)对2,2'-偶氮二异丁基脒二盐酸盐(AAPH)诱导的猪肾上皮细胞(LLC-PK1细胞)氧化应激的保护作用。海带由米曲霉在35±1°C下发酵72小时。将冻干的发酵海带用蒸馏水提取,提取液与乙醇混合后离心。上清液用各种溶剂进行分步分离。本研究使用BuOH组分是因为它在各种溶剂组分中具有最强的抗氧化活性。发酵海带的BuOH组分对AAPH诱导的LLC-PK1细胞损伤具有保护作用,可提高细胞活力,同时减少脂质过氧化的形成,并提高超氧化物歧化酶和谷胱甘肽过氧化物酶等抗氧化酶的活性。与完整的BuOH组分在相同浓度下显示的0.22±0.08 nmol丙二醛相比,BFLJ在100 μg/mL浓度下对脂质过氧化形成的抑制作用具有更高的值,为0.11±0.01 nmol丙二醛。此外,BFLJ处理可提高谷胱甘肽浓度。用100 μg/mL的BFLJ处理的细胞中谷胱甘肽浓度为1.80 pmol/L×10(5)个细胞。这些结果表明,BFLJ通过抑制脂质过氧化形成、提高抗氧化酶活性和谷胱甘肽浓度来保护LLC-PK1细胞免受AAPH诱导的细胞损伤。