食品科学

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竹盐酿造酱油对H2O2诱发LLC-PK1细胞氧化损伤的保护作用

宋家乐1,2,李贵节2,3,赵 欣2,3,*   

  1. 1.桂林医学院公共卫生学院,食品卫生与营养学教研室,广西 桂林 541004;2.重庆第二师范学院,功能性生态食品研究所,重庆 400067;3.重庆第二师范学院生物与化学工程系,重庆 400067
  • 出版日期:2015-05-15 发布日期:2015-05-11

Protective Effect of Bamboo Salt Soy Sauce on H2O2-Induced Cellular Oxidative Damage in LLC-PK1 Cells

SONG Jiale1,2, LI Guijie2,3, ZHAO Xin2,3,*   

  1. 1. Department of Food Hygiene and Nutrition, School of Public Health, Guilin Medical University, Guilin 541004, China;
    2. Institute of Functional Ecological Food, Chongqing University of Education, Chongqing 400067, China;
    3. Department of Biological and Chemical Engineering, Chongqing University of Education, Chongqing 400067, China
  • Online:2015-05-15 Published:2015-05-11

摘要:

目的:研究竹盐酿造酱油乙醇提取物对H2O2诱发猪肾近曲上皮小管细胞(pig proximal tubular cell)LLC-PK1氧化损伤的保护作用。方法:以不同质量浓度(10~200 μg/mL)的竹盐酿造酱油乙醇提取物预培养LLC-PK1细胞24 h后,换用含500 μmol/L H2O2的DMEM细胞培养液继续培养4 h建立细胞氧化损伤模型。四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法检测细胞生存率,硫代巴比妥酸比色法测定细胞内丙二醛(malondialdehyde,MDA)含量,比色法测定细胞内过氧化氢酶(catalase,CAT)、超氧化物歧化酶(superoxidedismutase,SOD)、谷胱甘肽过氧化物酶(glutathione peroxidase,GSH-Px)活性和谷胱甘肽(glutathione,GSH)含量。结果:经不同质量浓度竹盐酿造酱油乙醇提取物预处理24 h后,受损细胞的生存率上升,细胞内MDA生成量减少,ROS水平显著降低。同时,细胞内抗氧化酶(CAT、SOD和GSH-Px)的活性及其mRNA转录水平,GSH含量也较未经竹盐酿造酱油乙醇提取物处理的受损细胞增加。结论:竹盐酿造酱油乙醇提取物可以通过提高细胞内抗氧化酶系的活性和抗氧化物质GSH的含量而有效地对抗H2O2诱发的LLC-PK1细胞氧化损伤。

关键词: 竹盐酿造酱油, 氧化损伤, 抗氧化, LLC-PK1细胞

Abstract:

The aim of this study was to investigate the protective effect of ethanol extract from bamboo salt sauce (BSSE) on
H2O2-induced oxidative damage in porcine renal epithelial (LLC-PK1) cells. The LLC-PK1 cells were first incubated with
different concentrations of BSSE (10–200 μg/mL) for 24 h, and then exposed to H2O2 (500 μmol/L) for 4 h. MTT assay was
used to evaluate the cell viability. The cellular levels of reactive oxygen species (ROS), lipid peroxidation, and antioxidant
enzymes including catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and glutathione (GSH)
were measured. In addition, mRNA levels of these antioxidant enzymes were determined by RT-PCR assay. BSSE was able
to increase the cell viability, and also decrease the cellular levels of ROS, lipid peroxidation, as well as increase the activity
and mRNA expression of antioxidant enzymes when compared with those in control cells. These results suggest that BSSE
showed a protective effect against H2O2-induced oxidative damage in LLC-PK1 cells through inhibiting lipid peroxidation,
deceasing ROS levels, and increasing antioxidant system activities.

Key words: bamboo salt soy sauce, oxidative damage, antioxidant, LLC-PK1 cells

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